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Full text of "Commercial development ambient temperature starch hydrolysis"

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RENEWABLE ENERGY 
REPORT LIBRARY 

COMMERCIAL DEVELOPMENT 

AMBIENT TEMPERATURE 

STARCH HYDROLYSIS 



STATE DOCUMENTS COLLECTiOH 

MAR 1 iSG3 

MONT.V':.\ S-ATG LIBf7A 

VolZZ. 6ti-. AVE, 
HELEK'A, MOr iAi'4 r.-.^^,- 



Prepared for 
MONTANA DEPARTMENT of NATURAL RESOURCES and CONSERVATION 



PLEASE RETURN 



MONTANA STATE LIBRARY 



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HELENA, MONTANA 59620 



COMMERCIAL DEVELOPMENT 
lENT TEMPERATURE STARCH HYDROLYSIS 



Prepared by 

RENEWABLE TECHNOLOGIES, INC. 

P.O. Box 41 13, 630 Utah 

Butte, Montana 59702 

(406) 782-2386 



Prepared for 

Montana Department of Natural Resources and Conservation 

1520 East Sixth Avenue 

Helena, Montana 59620-2301 



Aval I abl e from 

Montana State Library 

1515 East Sixth Avenue 

Justice and State Library Builaing 

Helena, Montana 59620-2301 



This report was prepared under an agreement funded by the Montana Department of 
Natural Resources and Conservation. Neither the Department nor any of its 
employees makes any warranty, express or implied, or assumes any legal 
liability or responsibility for the accuracy, completeness, or usefulness of 
any information, apparatus, product, or process disclosed, or represents that 
its use would not infringe on privately owned rights. Reference herein to any 
specific commercial product, process, or service by trade name, traaemark, 
manufacturer, or otherwise, does not necessarily constitute or imply its 
endorsement, recommendation, or favoring by the Department of Natural Resources 
and Conservation or any employee thereof. The reviews and opinion of authors 
expressed herein do not necessarily state or reflect those of the Department or 
any employee thereof. 



ABSTRACT 

The objective of the two projects described in this report was to advance 
commercial development of an enzyme which could be used to eliminate starch 
cooking in alcohol fuel production. 

Cooking is necessary for the efficient action of enzymes now used to 
degrade (hydrolyze) starch to glucose prior to fermentation. Cooking accounts 
for up to one-third of the process energy used in alcohol production. Elimina- 
tion of cooking would substantially improve the economics of alcohol production. 

In previous work (DNRC Grant #RAE-82-1007) , RTI developed an enzyme which 
would efficiently degrade uncooked starch using innovative solid state culture 
technology (SSC) and a selected strain of mold. This enzyme is used in a simple 
one-step process for simultaneous hydrolysis and fermentation of starch in 
uncooked grain mash, Ambient Temperature Starch Hydrolysis (ATSH). 

Work described in this report was to scale up SSC technology to produce 
ATSH enzyme, improve production efficiency and evaluate commercial potential. 
Pilot plant work under Grant RAE-84-1044 developed the engineering date necessary 
to design larger scale culture systems and specify associated processing equip- 
ment. Laboratory work evaluated a number of variables affecting enzyme produc- 
tion efficiency resulting in a four-fold improvement. Grant RAE-85-1055 was a 
supplemental grant to develop improved mold culture substrates and SSC monitoring 
and control systems. 

Tests of the enzyme at commercial alcohol production plants demonstrated 
the efficiency and cost effectiveness of the no-cook process. 

All major objectives were met. Work culminated in preliminary design and 
equipment specifications for a first-stage commercial enzyme production plant 
designed to supply regional alcohol fuel producers. Economic analysis indicated 
that such a plant is profitable and could pay an attractive return on investment. 
The initial plant could be followed by expansion to serve national markets. 
RTI is working to raise private capital for construction of the plant. 

In addition to ATSH enzyme, RTI has obtained federal grant support for 
developing three additional products using SSC technology adapted from systems 
developed in DNRC-supported work. 



TABLE OF CONTENTS 



Page 



A. INTRODUCTION 1 

Starch Cookin>; 1 

Project History 4 

B. RESEARCH METHODOLOGY 8 

Laboratory Experimental Systems 8 

Culture Procedures 12 

ATSH Fermentation 13 

Enzyme Assays 15 

Computerized Data System 15 

Pilot Plant Systems 15 

C. RESULTS AND DISCUSSION 23 

1. Laboratory Results and Discussion 23 

Enzymology/ Biochemistry of Raw Starch Hydrolysis 24 

ATSH Fermentation 25 

hkjid Culture Parameters 28 

Culture Substrate 31 

Alternative Mold Strains 31 

Improved Culture Substrati> 32 

Monitoring and Control 33 

Conclusion 35 

2. Pilot Plant Results and Discussion 35 

3. Commercial Demonstration 37 

Dillon, Montana Test 38 

ADM Test 40 

D. SYSTEM DESIGN 43 

E. ECONCMICS 50 

1. Market Analysis 50 

2. Cash Flow Analysis 54 

F. GRANT ADMINISTRATION 59 

1. Work Schedule 59 

2. Budget 62 

G. RESULTS AND CONCLUSIONS 65 

1. Major Findings and Recommendations 65 

2. Permits, Licenses, and Authorizing Agencies 66 

3. Additional Development 68 

APPENDIX 1 - Computer Software Monitoring and Control System 
APPENDIX 2 - Assay Procedures and Instrumentation 
APPENDIX 3 - Drawings and Schematics 
APPENT)IX 4 - Mass and Energy Balance 
APPENDLX 5 - Financial Analysis 



LIST OF FIGURES 



Page 



Figure 


1. 


Figure 


2. 


Figure 


3. 


Figure 


4. 


Figure 


5. 


Figure 


6. 


Figure 


7. 


Figure 


8. 


Figure 


9. 


Figure 


10. 


Figure 


11. 


Figure 


12a, 


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Figure 


14b, 


Figure 


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Figure 


16. 


Figure 


17. 


Figure 


18. 


Figure 


19. 


Figure 


20. 



Fuel Alcohol Production Process 2 

Comparison of Conventional vs. ATSH Process for 

Alcohol Production 4 

Reactor Instrumentation and Control System: 

Simplified Schematic 10 

Photographs of Culture System with Improved Monitoring 

and Control 11 

7 Liter Fermentation System 14 

200 Liter Fermenter 14 

Glass Column 17 

Holo-Flite Processor 18 

Glass Columns with Auger System 19 

Heat Exchanger Plates 19 

Concentric Screen Reactor Internals 20 

Tray 21 

Reactor 21 

Lifting Flight Reactor 22 

ATSH Fermentation - Barley Starch 26 

ATSH Fermentation - Corn Starch 27 

ATSH Fermentations: 7 Liter Bench Fermenters, Reduced 

Enzyme Dose Rates on Barley 29 

ATSH Fermentations: 7 Liter Bench Fermenters, Reduced 

Enzyme Dose Rates on Whole Corn 30 

ATSH Fementation Scale-up Tests 39 

ATSH Feraentations from ADM Tests 42 

Project Time Line - Grant Agreement No. RAE-84-1044 60 

Project Time Line - Grant Agreement No. RAE-85-1055 61 



LIST OF TABLES 



Table 


2. 


Table 


3. 


Table 


4. 


Table 


5. 


Table 


6. 



Page 

Alcohol Fuel Production Companies Which Have Contacted 

RTI Regarding ATSH Enzyme 52 

Enzyme Production Analysis 55 

Statement of Projected Results of Operations and Cash Flows... 58 

Budget for Grant Agreement No. RAE-84-1044 63 

Budget for Grant Agreement No. RAE-85-1055 64 

List of RTI Biofuel Contracts for Solid State Culture 

Development 64 



A. INTRODUCTION 

Alcohol could reduce Montana's reliance on fossil fuels. It can be used 
to replace petroleum-based or lead-based octane boosters in gasoline and can 
be used by itself as fuel in specially modified engines. Conversion of Montana 
grain crops to alcohol is a value-added processing industry that benefits the 
Montana economy by providing for four product markets, creating employment and 
creating a new tax base. 

The current process for converting grain starch to alcohol is a multi-step 
process as depicted in Figure 1. Energy inputs are required in feedstock pre- 
paration, cooking, distillation and drying of distiller's grains produced as a 
coproduct and marketed as high protein livestock feed. Research and development 
work conducted by RTI under the two contracts described in this report was 
directed toward eliminating the cooking step from alcohol fuel production. 
This would significantly improve the energy efficiency and economics of alcohol 
fuel production. 

Starch Cooking 

Starch is a polymer of glucose molecules bound together in chains. In 
grain starch, molecules are contained in microscopic starch granules which are 
insoluble in cold water. Starch conversion or "cooking" is a multistep process 
in which starch is broken down to glucose. Glucose is then fermented by yeast 
to ethanol. (Yeast used in alcohol production cannot directly ferment starch.) 
In the current enzymatic starch conversion process, a starch/water slurry or 
mash is cooked at boiling or above and then treated in separate steps with two 
enzymes. Cooking breaks up or gelatinizes the starch granules, rendering the 
starch susceptible to the action of the enzymes. The first enzyme (alpha 
amylase) is added when the mash is about 200°F. This enzyme breaks the long 



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starch chain into shorter, soluble glucose chains. This step is generally called 
"liquefaction." The second enzyme (glucoamylase) is added after the mash is 
cooled to about 140°F. This enzyme releases glucose molecules from the ends of 
the short chains. The cooking step is essential for the efficient and economical 
use of these enzymes. In small alcohol plants such as those in Montana, cooking 
is a batch process in which a tank of mash is heated to near boiling, treated 
with enzymes and then cooked and fermented. In larger plants, continuous "jet 
cookers" are used. 

The energy and capital cost of cooking varies depending on plant scale, 
feedstock and process used, but generally accounts for about 30% of overall 
plant energy use and 20 to 40% of capital cost. 

Elimination of cooking would require an enzyme or combination of enzymes 
that would efficiently degrade starch contained in starch granules. The 
objective of RTI's research and development work for DNRC was to develop such 
an enzyme and define the process for its use in alcohol fuel production. 

RTI has successfully developed pilot scale technology for producing an 
enzyme preparation to degrade granular starch and demonstrated the enzyme in 
converting grain starch to alcohol. Conversion of starch to glucose and fer- 
mentation of glucose to alcohol is a simple one-step process that takes place 
entirely at fermentation temperature. RTI calls this process Ambient Tempera- 
ture Starch Hydrolysis (ATSH). Alcohol production using the conventional 
cooking process and the ATSH process are compared in Figure 2. In the ATSH 
process, cooking is eliminated entirely. Standard fermentation tanks can be 
used for the process, eliminating both the operating and capital cost of cooking 
systems. Overall conversion rate and efficiency is equivalent to conventional 
processes and enzyme costs are similar. This report details test results. 



preliminary design for an enzyme production plant and the economics of enzyme 
production and use. 



COr;VErJTI0MAL ENZVM-IJ ■.-.,. 'ATSH" ENZVXE tor Alcohol Prcducticn |j 




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Figure 2. Comparison of Conventional vs. ATSH Process for Alcohol Production 



Project History 

Development of the ATSH enzyme was funded by three grants totaling $480,000 
from the Montana Department of Natural Resources and Conservation (DNRC) under 
the Renewable Alternative Energy Grants Program. Additional support of $13,000 
was provided by the Montana Department of Agriculture for work on questions 
related to co-product distiller's grains. RTI contributed over $60,000 in 
matching funds. RTI has used the basic technology developed through DNRC support 
to obtain over $600,000 in federal grants for research and development work on 
three other biological products, all of which have significant commercial poten- 
tial and could be produced in Montana. 

The initial DNRC grant (Contract RAE 82-1007) funded laboratory research 
to test the feasibility of developing an enzymatic process for converting 
uncooked starch. This first phase, completed in September 1983, was successful. 
Production of the ATSH enzyme resulted from two breakthroughs. The first was 

4 



identifying a mold which produced an enzyme complex with the necessary charac- 
teristics. The second was development of innovative technology for culturing 
the mold. This first phase of research defined development of the culture 
system as the central problem in commercial development of the ATSH enzyme. 

Conventional technology for producing mold derived-enzymes is submerged 
culture or liquid state culture (LSC). In LSC, the mold is cultured as a sus- 
pension in a water-based nutrient solution generally in tanks of 10,000 to 
100,000 gallons. In nature, mold normally grow as surface cultures on moist, 
solid substrates. Liquid environments are normally occupied by bacteria. 
Although LSC is not a normal environment, strains of mold used for industrial 
enzyme production have been adapted or mutated for use in LSC. In LSC, process 
conditions such as temperature, nutrient concentrations and agitation can be 
readily controlled. 

The ATSH enzyme is produced by culturing the selected mold strain in solid 
state culture (SSC). In SSC, the mold is grown on the surface of moist solid 
nutrients. Solid state culture has been used in oriental beverage and food 
production for centuries. Solid culture of mold are used in production of 
sake (rice wine), miso and soy sauce. Traditional Japanese solid mold cultures 
are referred to as Koji. In the mid-1940s, ko j i technology was adapted to 
produce enzymes which were used in alcohol production. In large scale, SSC 
process conditions proved very difficult to control and, with the end of war 
time economics, SSC was abandoned in the U.S. in favor of LSC. 

The initial research led to development of a laboratory experimental systen 
in which the mold was grown in a packed bed reactor using processed barley as 
a nutrient and bed support. The use of SSC in combination with the selected 
mold strain resulted in production of starch degrading enzymes which are 
functionally and probably structurally different than enzymes produced using 

5 



LSC. The solid culture enzymes efficiently degraded uncooked starch at fermen- 
tation temperature of 35*'C at a pH of 3.5. A simple process was developed for 
mixing the mash, enzyme and yeast In a simultaneous conversion of starch to 
glucose and fermentation of glucose to alcohol. Preliminary economic evalua- 
tions indicated that the enzyme could be produced at a cost competitive with 
existing commercial enzyme preparations used in alcohol production. 

Based on these results, RTI was awarded additional funding (DNRC Grant 
RAE-84-1044) to support scale-up through design, construction and testing of 
pilot plant facilities. Specific objectives were to: 

Obtain the engineering and economic data to provide the basis 
for subsequent financing, design and construction of a small 
commercial ATSH enzyme production plant. 

Continue to improve the economics of ATSH enzyme production. 

Work under the contract began in May 1984. 

In April 1985, RTI received supplemental funding (UNRC Grant RAE-85-1055) 
for additional work on two crucial aspects of SSC development for ATSH enzyme 
production. These were feedstock processing for preparation of mold culture 
substrate and improved monitoring and control. Specific objectives in these 
two areas were: 

Feedstock Processing 

Identify an alternative form of processed barley with Improved 
culture substrate characteristics. 

Determine the availability of equipment for feedstock processing 
Including the necessity for adaptation and modifications to fit 
ATSH enzyme production requirements. 

Evaluate the opportunity to recover co-products for sale as live- 
stock feed and determine the contribution of co-products to over- 
all plant economics. 

Develop preliminary designs and experiment cost estimates for the 
feedstock processing component of an ATSH enzyme production plant. 



Improved Culture Monitoring and Control 

Improve enzyme production efficiency. 

Determine culture monitoring and control requirements for commer- 
cial ATSH enzyme production. 

Develop preliminary design and equipment cost estimates for 
monitoring and control systems in an ATSH enzyme production plant, 



B. RESEARCH METHODOLOGY 

This section describes the laboratory experimental systems, culture 
procedures, analytical procedures, and pilot plant systems used In the develop- 
ment of ATSH enzvme. 

Laboratory Experimental Systems 

Two experimental systems were used. Both used either 250 or 500 cc clear 
plastic columns for culture reactors. Initially the system consisted of 15 
columns Immersed in a temperature-controlled water bath. Each column had 
individual controls for a flow of humidified air and continuous temperature 
monitoring. 

Work with the initial laboratory experimental system and pilot plant tests 
indicated a need for improved monitoring and control capability. Under Con- 
tract RAE-S5-1055, a second generation laboratory experimental system was 
constructed. This system was based on Individual computerized monitoring 
and control for 10 individual culture reactors. Continuous monitoring capa- 
bilities include temperature, airflow rate, air pressure, humidity, and con- 
centration of oxygen and carbon dioxide. Both inflow and outflow air can be 
sampled and analyzed. Control capabilities include variations in temperature, 
airflow rate and pressure, humidity and atmosphere composition. The concen- 
tration of oxygen, carbon dioxide and nitrogen can be controlled Individually 
for each culture. Computer actuated electronic valves are used for gas sampling 
and gas blending and for controlling the airflow regime. Airflow through the 
culture can be continuous with exhaust to atmospheric pressure, continuous 
with controlled back pressure or intermittent with time variable on/off cycles. 
As with the previous sytem, a water bath was used for temperature control. 
The entire system was placed in a temperature controlled cabinet to ensure 



proper calibration of sensors and avoid condensation in lines and valves. 
Figure 3 is a schematic drawing of the system, and Figure 4 shows photographs 
of the system. 




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A. Culture Test Stand 



Computer Monitoring and 
Control System 



Figure 4. Photographs of Culture System with Improved Monitoring and Control 



11 



Portions of the design and computer software were developed by MultlTech, 
Inc. of Butte under subcontract to RTI. A complete description of computer 
software Is included in Appendix 1. 

Development of this sophisticated laboratory SSC experimental system 
provided important insight into metabolisms and enzyme production by the mold. 
This was crucial to development of pilot plant reactor design and control sys- 
tems. One important element in the ability to design and effectively control 
large commercial SSC systems is the application of state-of-the-art sensors and 
computerized monitoring and control systems. 

Culture Procedures 

Mold strains used in laboratory and pilot plant tests were maintained as 
pure cultures on agar slants. Slant cultures were used to inoculate solid 
cultures grown under conditions which enhanced spore (conidia) production. 
These spore cultures were dried, ground and stored at 4°C for use in starting 
experimental cultures. This procedure represents an efficient procedure for 
maintaining cultures and preparing inoculant in commercial SSC enzyme production. 

Enzyme production cultures utilized solid, barley-based particles as sub- 
strates. Physical and biochemical substrate characteristics were important 
variables and substrate preparation procedures varied. Generally particles of 
processed barley were soaked in a nutrient solution and steamed. When cooled, 
the substrate was inoculated with a freshly prepared slurry of spores and loaded 
into the culture reactor. Laboratory and pilot plant procedures were essentially 
identical. 

Enzyme production cultures were grown in culture reactors for variable 
periods, generally 72 hours or to the points defined by monitoring data on 
temperature or oxygen consumption. At the end of the culture period, the 

12 



entire culture, including cells, extracellular protein and residual culture 
solids, was dried to about 5% moisture and milled to produce a coarse powder. 
This crude, dried whole culture was used as ATSH enzyme. Economic evaluations 
indicate that this procedure is most economical in commercial production of the 
enzyme. As a result, there was no further enzyme purification. 

ATSH Fermentation 

ATSH fermentation refers to the process in which starch mash, enzyme and 
yeast are mixed in a one-step conversion of starch to glucose and simultaneous 
fermentation of glucose to ethanol. 

ATSH fermentations were conducted at laboratory and pilot scale using both 
barley and corn as feedstocks. 

A standard laboratory fermentation of 25% grain in 100 g total mash was 
used as an assay of enzyme preparations. This procedure provided the best 
comparative analysis of experimental enzyme preparations. 

Pilot scale fermentations were run in 7 liter and 200 liter fermenters. 
Whole ground barley, whole milled corn and purified corn starch were used in 
pilot fermentations with variable starch concentrations and enzyme dose. 
Pilot fermenters were designed as models of large ethanol fermenters and 
included control of temperature, variable agitation and pH. Fermentation 
assay capabilities included ethanol concentration, glucose concentration, 
total reducing sugars, yeast count, bacterial contamination, and residual 
starch. Figures 5 and 6 are photographs of 7 liter and 200 liter fermenters. 

Yeast used in ATSH fermentations was a commercial distillery yeast mar- 
keted by Gist Brocades under the trade name Fermiol. Yeast were maintained 
and cultured using standard techniques. 



13 







Figure 5. ' Liter Ferraentation System Consisting of 7 Liter Polycarbonate 

Fermenter Vessel in a Temperature Controlled Bath, Variable Speed 
Stirring Motors with Torque and rpra Monitoring and pH Monitoring 







1% 0k 




Figure 6. 200 Liter Fermenter 
14 



ATSH fermentations are started with a 1 to 3% v/v inoculation of yeast 
culture. Initial pH is 3.5 to inhibit bacteria. 

Enzyme Assays 



A set of enzyme assay procedures was developed to measure different starch 
degrading activities in the crude ATSH enzyme preparations. Different enzymes 
were determined by using assay substrates selective for glucoamylase, alpha 
amylase and debranching activities. Assay procedures and instrumentation used 
in the work are described in more detail in Appendix 2. 

Computerized Data System 

A data base program was developed to store and evaluate information on 
each enzyme production test including culture conditions, variables tested, 
fermentation results, and enzyme assay values. The final version of the data 
base had graphing capability for plotting results and basic statistical func- 
tions for correlating results. Principal statistical evaluations were to run 
correlations between fermentation results and enzyme assay values. 

Pilot Plant Systems 

Pilot plant experimental systems includes SSC mold culture reactors and 
processing equipment. Design of efficient SSC reactors which could be con- 
trolled at commercial scale was the principal technical problem addressed in 
these projects. 

During the course of the projects, 7 different pilot plant scale culture 
reactors were designed, constructed and tested. A number of the reactors 
were built for a specific set of experiments designed to gather data on a 
specific design or operating problem. Information from these tests was in- 
corporated into the final reactor design described in Section G. Photographs 

15 



of different reactors are shown in Figures 7-13. Reactors included: 

Figure 7. Glass columns 6" x 30" equipped with internal heat exchangers 
for heat exchange and temperature control experiments. 

Figure 8. "Hollo Flight" auger system consisting of twin counter rotating 
augers used for material handling experiments. 

Figure 9. Glass columns 6" x 50" equipped with internal auger used for 
materials handling experiments. 

Figure 10. Heat exchanger plate system 24" x 24" and 24" x 60". Two 

plates set at variable distances with mold culture between the 
plates were used for heat transfer experiments and preparation 
of enzyme for pilot plant and commercial demonstration tests. 

Figure 11. Concentric screen reactor contained culture between two cylin- 
drical screens. Airflow was from the outside with exhaust 
through the center. This design was used for airflow experiments. 

Figure 12. Tray reactor system with 18" diameter screen bottom trays in a 
vertical cylinder. Some trays were designed with heat exchange 
circulation incorporated in the screen. This system was operated 
with variable depths of substrate bed for temperature and air- 
flow experiments. This system was also used with oxygen control 
and monitoring systems for experiments on atmospheric composition. 

Figure 13. Lifting flight reactor with 4 hollow center vertical augers in an 
18" X 3b" vertical reactor. The reactor was equipped with high 
volume airflow and a temperature actuated spray system for cooling. 
Augers were designed to lift and gently mix the mold culture. 

Several types of equipment were used in support of pilot plant tests and 
in experiments to define process conditions and equipment design specifications 
for substrate processing, materials handling, reactor control, culture drying, 
and milling in commercial ATSH enzyme production. Samples of material were 
also sent or taken to equipment vendors for tests of potential types of pro- 
cessing equipment. Pilot plant equipment included: a 24 kg balance, drum roller 
and a pressure vessel for substrate preparation; several different types of 
drvers; compressors; pressure regulators and flow controllers for air delivery; 
data logger; oxygen monitor; humidity sensors for process control; and a mill. 

Vendor equipment tests included several types of substrate preparation 
equipment and several types of dryers. 

16 




Figure 7. Glass Column 



17 




Figure 8. Holo-Flite Processor 



18 







Figure 9. Glass Columns with Auger System 




Figure 10. Heat Exchanger Plates 



19 




Figure 11. Concentric Screen Reactor Internals 



20 




Figure 12a. Tray 




Figure 12b. Reactor 
21 




Figure 13. Lifting Flight Reactor 



22 



C. RESULTS AND DISCUSSION 

Results and discussion will be presented in three sections: 1) laboratory, 
2) pilot plant and 3) commercial demonstration. 

1. Laboratory Results and Discussion 

Laboratory experimental programs in both contracts were designed to improve 
enzyme production efficiency. This is defined by enzyme dose rate, enzyme 
recovery and reactor productivity. Dose rate is the amount of enzyme required 
in ATSH fermentation to meet selected specifications for rate, conversion effi- 
ciency and alcohol concentration. These specifications can vary in commercial 
alcohol production based on feedstock, plant design scale and operating schedules. 
For experimental programs, dose rate was defined as the amount of enzyme required 
to achieve at least 90% of theoretical starch to ethanol conversion efficiency 
with final alcohol concentrations of at least 9% v/v in 64 hours or less. Dose 
rate is expressed as weight percent of enzyme in total weight of mash. 

Enzyme recovery is the amount of enzyme recovered from a given amount of 
culture substrate expressed as a weight-based percentage. Reactor productivity 
is the amount of enzyme produced from a given reactor volume and is a function 
of culture time, recovery rate and enzyme concentration. The most important 
measure of ATSH enzyme economics is dose rate. 

The original proposals listed six laboratory tasks organized in six mile- 
stones in Contract RAE-84-1044 and five laboratory tasks in four milestones 
for Contract RAE-85-1055. For clarity, results will be presented according to 
the following outline: 

RAE-84-1044 

Enzymology/ Biochemistry of Raw Starch Hydrolysis 
ATSH Fermentation 



23 



Culture Parameters 

Culture Substrate 

Mold Strains 
RAE-85-1Q55 

Culture Substrate 

Monitoring and Control 
These categories reflect the emphasis of work as it was actually conducted in 
response to results and technical questions important in commercial development 
of ATSH enzyme. Laboratory work under the two contracts included over 1300 
Individual mold cultures. Individual cultures were evaluated in shake flask 
ferraentlons and enzyme assays. Selected preparations were used for enzymology 
studies, 7 liter bench fermentations and commercial demonstrations. 

RAE-84-1044 

Enzymology/Biochemistry of Raw Starch Hydrolysis 

The enzymology and biochemistry of raw starch hydrolysis is complex and 
not completely understood by either RTI or other research groups. Work in this 
area was conducted with two objectives. The first was to develop a set of 
rapid enzyme assays to predict enzyme performance in ATSH fermentation. The 
second was to better understand raw starch hydrolysis as a basis for improving 
enzyme production efficiency. 

Standardized fermentations provide the best basis for predicting perfor- 
mance of ATSH enzyme. However, this requires two days. Enzyme assays provide 
rapid culture analysis and quality control in commercial production. Assays 
were developed for alpha amylase, glucoamylase and debranchlng activity. 
Correlations between assays and fermentation results were very high for mold 



24 



strains and culture conditions used during the first year of work. Correlations 
were poor with later changes in strain, substrate and control parameters. Sig- 
nificant improvements in dose fermentation rate have been achieved without 
corresponding increases in enzyme assay values. Further work will be required 
to develop reliable quality control assays for commercial production. 

Studies of raw starch hydrolysis included comparisons between ATSH enzyme 
and commercial preparations, absorbtion studies, hydrolysis of different types 
of starch, determination of pH optima, and microscope studies. Results of this 
work clearly showed functional differences between ATSH enzyme and commercial 
amylases. Results suggest differences in protein structure. Differences 
include pH optima, starch absorption patterns, hydrolysis rates on different 
starches, and conversion efficiency. ATSH enzyme rapidly and completely 
hydrolyzes raw starch. Conventional amylases show slower reaction rates in 
raw starch hydrolysis and typically convert less than 60% of available starch. 
Figures 14 shows microscope photographs of raw starch hydrolysis by ATSH enzyme. 

ATSH Fermentation 



ATSH fermentations were run in 250 ml shake flasks, 7 liter bench fermen- 
ters and 200 liter pilot fermenters. Commercial demonstrations were also 
conducted in 4 liter bench fermenters and 1200 gallon industrial fermenters. 

Shake flasks were used as a screening procedure for preliminary definition 
of fermentation variables including yeast dose, acid concentration, enzyme 
concentration and starch sources. Results from shake flasks were confirmed in 
scale up at 7 liter and 200 liter. 

Shake flask fermentations were also routinely employed as an assay proce- 
dure to compare different enzjmies and for correlations with enzyme assays. 



25 



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Figure 14a. ATSH Fermentation - Barley Starch 



26 



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Figure 14b. ATSH Fermentation - Corn Starch 



27 



Bench scale fermentations were employed to define optimum fermentation 
conditions as a means to reduce enzyme dose and improve conversion efficiency. 
Agitation is the most important variable in scale-up of ATSH fermentation. 
Agitation typical of conventional pilot fermentation systems inhibited ATSH 
fermentation. Inhibition apparently results from effects of shear forces on 
binding of enzyme to starch granules. Optimum agitation in 7 liter fermenters 
was determined to be intermittent for 2 minutes per hour. Agitation speed and 
blade design used were just sufficient to completely mix the mash. 

Figures 15 and 16 show typical results for 7 liter fermenters of barley 
and corn at different enzyme dose rates. Results from 7 liter bench fermenters 
accurately predicted results in commercial demonstration tests. 

Mold Culture Parameters 

Variables which affect mold growth and enzyme production in SSC include 
inoculation, temperature, airflow rate, humidity, culture time, and biochemical 
and physical substrate characteristics. Physical substrate characteristics 
proved to be very important and will be discussed below. This section summar- 
izes results of experiments to monitor and control other culture variables. 

Inoculation procedures were developed for use in laboratory, pilot plant 
and commercial operations. Culture procedures which promote spore (conidia) 
formation in SSC were developed. Spore cultures were dried and milled to pro- 
duce a stable spore preparation which is used to inoculate production cultures. 
For inoculation, spore cultures were slurried In water and mixed with substrate, 

Systems for monitoring and control of temperature, airflow and humidity 
were described in Section B. Systems constructed initially as part of Contract 
RAE-84-10AA were used to establish baseline conditions for culture uniformity. 



Is 



|2 



r 

X 

S3 



:W 





\ 



xww 


xww 


rfMHt 


WWJE-H 


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OiAK 


fflfiK 


I 


^ 




Experiments were then conducted to define conditions for inoculation rates, 
airflow, temperature, and culture time. This was followed by experiments to 
evaluate addition of organic and inorganic nutrient supplements. A low-cost 
supplement was developed which significantly improves enzyme production. 

Culture Substrate 

Physical substrate characteristics proved to be an important variable in 
enzyme production efficiency and critical to design of pilot and commercial 
production systems. Important characteristics include density, packing char- 
acteristics, moisture retention, surface area, and utilization by the mold. 
Substrate utilization is a critical economic factor affecting enzyme recovery, 
unreactive solids in enzyme preparations, feedstock requirements and enzinne 
concentrations. Initial substrate evaluations included a variety of commer- 
cially available forms of processed barley which were further processed to 
varying degrees in laboratory experiments. This initial work was unsuccessful 
in identifying an improved substrate. This work did define the importance of 
physical substrate characteristics leading to further work in this area as 
part of Contract RAE-85-1055. 

Alternative Mold Strains 



Basic research on ATSH enzyme production had compared a limited number of 
mold strains which showed a high degree of variability in producing high con- 
centrations of the enzymes with the desired characteristics. Work in Contract 
RAE-84-104A compared an additional 22 strains individually by enzyme assay and 
standard fermentations. Enzymes from cultures of different strains were also 
mixed and tested in standardized fermentations as a means to reduce dose rate. 
All were tested in 2 strain mixtures. The best were tested in 3 strain mixtures 



31 



This work was successful in identifying a strain which produced significantly 
higher concentrations of raw starch -active enzyme allowing reduced dose rates. 
The strain is also superior in temperature control characteristics. 

Mixed strain enzyme preparations resulted in only small improvements in 
dose rate. Results did not appear to justify the additional complexity and 
expense of using two separately cultured organisms in commercial production. 

RAE-85-1055 



Improved Culture Substrate 

Based on initial substrate tests, a more exhaustive evaluation of physical 
substrate characteristics was undertaken. This work was conducted in four 
parts: an initial survey of starch processing equipment manufacturers, prelim- 
inary tests in which samples were sent to selected manufacturers for processing, 
on-site substrate production tests, and laboratory evaluation of substrates in 
ATSH enz3Tne production cultures. Substrates were evaluated for packing charac- 
teristics, void space, surface area, starch utilization, and enzyme recovery. 

The initial survey included 34 companies of which 16 responded with product 
literature and information in follow-up telephone discussions. Samples of 
barley starch were sent to 8 companies representing each of the basic types of 
process equipment. Processed samples were received from 6 companies for 
laboratory testing. Based on laboratory results, tests were conducted on-site 
at two companies using their demonstration facilities and staff assistance. 
These tests produced quantities of substrate for more thorough lab tests and 
provided valuable experience in evaluation of the equipment. Laboratory tests 
of commercially processed substrate samples included a variety of additional 
process steps to add moisture, vary density and affect physical strength. 
Substrate variables were tested in over 300 individual cultures. 

32 



Tests did not show any increase in enzyme production efficiency using 
substrates obtained from equipment tests. However, significant improvements 
were obtained when laboratory process steps learned in this work were applied 
to substrates previously tested in RAE-84-1044. As a result, dose rates in 
ATSH fermentations were reduced by half. 

Substrate evaluation also included evaluation of by-products. The first 
step in preparing barley for use as culture substrate is pearling to remove 
hull. This produces a waste which can be marketed as cattle feed for $20 to 
$40 per ton. Market price varies with protein content and the price of grain. 
Economic evaluations include by-product value based on cattle feed markets. 
Pearling waste may also have application in formulating culture substrate in 
production of cellulose degrading enzymes using SSC technology. In work 
sponsored by the U.S. Department of Energy (U.S. DOE), RTI has successfully 
adapted SSC technology to production of cellulose degrading enzyme. If used 
commercially by RTI in this application, value added to pearling waste would 
range from $170 to $450 per ton. 

Monitoring and Control 

This work included design and construction of a laboratory experimental 
system with much more sophisticated monitoring and control capability than 
previous systems. Design of the system is described in Section B. 

This system was used for 31 tests for a total of over 300 individual 
cultures. Tests evaluated: metabolic rates; metabolic heat production and 
temperature control; substrate utilization and enzyme recovery; culture time; 
and effects of controlled atmosphere. Two different strains were used in these 
tests and a range of substrate formulations. 



33 



Use of this system did not provide significant iraprovements in enzyme 
concentration or dose rate. Results provided scale-up data for 1) reactor 
design and control and 2) control of spore formation. Baseline information 
was developed for use in pilot plant tests and design of commercial reactors 
and control systems. Information on metabolic rates derived from oxygen and 
carbon dioxide monitoring data provided a quantitative basis for setting oper- 
ating parameters in larger SSC cultures. Lab data were used to predict oxygen 
requirements, carbon dioxide evaluation, moisture balances, and temperature 
profiles of larger SSC cultures. Predictions were confirmed by pilot plant 
tests, providing a quantitative rather than empirical basis for design of 
commercial SSC reactors and processing equipment. 

Data was also developed in control of spore formation by the organism. 
The presence of mold spores in ATSH enzyme preparations were of concern for 
two reasons. Spores are viable, so potential competitors could culture RTI 
organisms from enzyme preparations. This is of particular concern to potential 
private investors. The second concern is in marketing. Although the spores 
do not adversely affect performance of the enzyme, several alcohol companies 
interested in ATSH enzjnne expressed concern because of the black color and the 
need to control airborne spores. Spores could be removed from ATSH enzyme 
preparations by mechanical process steps; however, this would add additional 
capital and operating costs. Spores could also be controlled at the point of 
use by standard dust collection equipment. However, the best option was to 
prevent spore formation in the cultures. 

The sophisticated monitoring and control capability allowed testing of a 
wide range of variables affecting spore production. As a result, operating 
parameters were defined which completely suppress spore production without 
significant effects in enzyme concentrations or recovery rates. Pilot plant 

34 



tests designed from laboratory data were successful in producing spore-free 
ATSH enzyme preparations. 

Conclusion 

Laboratory work under Contracts RAE-8A-1044 and RAE-85-1055 achieved sig- 
nificant improvements in enzyme concentration, reduced dose rates and economics. 
Dose rates were reduced from 1.5% of mash weight at the time work started to 
0.375% for typical preparations. With some preparation, acceptable fermenta- 
tion rates and yields for barley fermentation in small scale plants have been 
achieved at .25% dose rates. Economic analysis discussed in Section H is 
based on dose rate, enzyme recovery and substrate formulation results achieved 
in laboratory work and confirmed by pilot tests. 

2. Pilot Plant Results and Discussion 

Eighty-three pilot plant experiments were conducted. Seven different 
reactor configurations were used with several different sizes in each configu- 
ration. Reactor capacities varied from 10 liters to 100 liters. 

The original experiments were focused on defining various physical charac- 
teristics and control parameters affected by reactor system scale-up. Different 
reactor configurations were designed to test various characteristics. Later 
experiments were designed to test optimum operating conditions defined by 
laboratory experiments and economic analysis. Results are summarized in the 
following categories: 

a. Materials Handling 

b. Control Parameters 

c. Enzyme Production Efficiency 

d. ATSH Fermentations 

e. Packaging 

35 



a. Materials Handling 

Substrate physical characteristics are critical to reactor design. Loading 
characteristics, physical strength, void space, particle porosity, water reten- 
tion, surface area, and starch availability affect enzyme growth. They are the 
limiting properties in maximizing reactor capacities. Various reactors were 
designed to test individual and multiple properties under different operating 
conditions. Whenever a substrate was changed, these characteristics were re- 
evaluated. The evaluations were used to optimize the final pilot plant reactor 
design. While some substrates had better physical properties, the enzyme was 
not as good. Barley flakes combined with barley straw in a 9:1 ratio by weight 
provided the best economic answers while satisfying the physical requirements. 

b. Control Parameters 

Recovery rate, dosage rate and substrate utilization have the most effect 
on the process economics. Laboratory experiments were used to determine the 
optimum conditions to achieve the most economical operation. Pilot plant 
experiments were run using the laboratory conditions to control bed tempera- 
ture, airflow, final moistures and spore formation. The critical control 
parameters were bed temperature and airflow. 

c. Enzyme Production Efficiency 

Pilot plant reactors typically ran 72 hours. Dry recovered enzyme was 60% 
of the dry feedstock on a pound/pound basis (recovery rate). Feedstock was 90% 
by weight barley flakes and 10% barley straw. Dosage rates were 0.0041 pounds 
of dry ATSH enzyme per pound of 25 percent solids barley mash. This is based 
on a typical barley mash where one gallon of ethanol is produced from 10 gal- 
lons of mash. 



36 



d. ATSH Fementations 

Fermentations using pilot plant enzyme were done in the laboratory. They 
required .0041 pounds of ATSH enzyme for each pound of barley mash. This is 
equivalent to .375 pounds of ATSH enzyme per gallon of ethanol produced. Some 
laboratory-produced ATSH enzyme worked well at .25 pounds per gallon of ethanol. 
With additional work, it may be possible to further reduce the dosage of the 
pilot plant enzyme. However, it is still economical at .375 pounds ATSH enzyme 
per gallon of alcohol and a 60% recovery rate from the SSC reactor. 

g. Packaging 

It will be necessary to dry, grind and package the enzyme. The enzyme 
would probably be packaged in 50-pound paper bags or drums. Different users 
may want a different size or container. 

Recent experiments were successful at eliminating spores while maintaining 
enzyme quality. Since no spores are formed, controlling spores would not be a 
problem. A wet scrubber would be installed on the grinding/packaging operation 
to control dust. 

3 . Commercial Demonstration 

Two commercial demonstrations of ATSH enzyme in alcohol production were 
conducted. The first was in a small alcohol plant in Dillon, Montana. The 
second was two sets of tests conducted at the laboratory of Archer Daniels 
Midland Company (ADM) in Clinton, Iowa. These tests represent the extremes 
in alcohol production scale and technology and demonstrated that ATSH enzyme 
has potential markets in the entire U.S. alcohol industry. Tlie plant in 
Dillon produces about 500,000 gallons of alcohol per year from dry milled 
wheat or barley. Cooking is 1200 gallon batch tanks which serve for both 



37 



cooking and fermentation. The plant purchases enzymes from commercial enzyme 
companies. 

ADM is the largest fuel alcohol producer in the world with a total capa- 
city of 365 million gallons of alcohol per year from four midwest plants. ADM 
produces alcohol as one product of an integrated wet milling process. Other 
products include purified corn starch, glucose, corn syrups, high fructose 
corn syrup, corn oil, and a variety of protein products marketed principally 
as livestock feeds. ADM employs continuous jet cooking technology for starch 
hydrolysis. For fermentation, 250,000 to 500,000 gallon tanks are employed in 
a continuous cascade system. Enzymes are produced in-house for all operations 
at the Clinton, Iowa plant. Results from these two demonstrations are discussed 
below. 

Dillon, Montana Test 

Prior to the commercial test, RTI conducted four pilot plant fermentations 
to determine any scale-up problems with ATSH fermentations. Physical operating 
parameters and recipes were duplicated from the best 7 liter (1.85 gallon) 
fermentation tests. With these variables held constant, the major scale-up 
factor was fermenter size, design and geometry. 

The ATSH enzyme used in the scale-up tests was pooled from pilot and 
laboratory cultures. All tests used barley mash. The pilot plant tests were 
conducted with RTI's 200 liter (52. 8A gallons) fermenter. The commercial 
demonstration test was conducted with a 1200 gallon (4542 liter) fermenter at 
Southwest Montana Alcohol Plant in Dillon, Montana. The ethanol values from 
the 7 liter, 200 liter and 4500 liter fermenters are shown in Figure 17. 
These results demonstrate that the ATSH process can be scaled up from 7 liters 
to 4500 liters. There should be no reason why the ATSH process cannot be 

38 







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z 


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scaled up to very large industrial fermenters as long as operating parameters, 
mash concentrations, pH, and enzyme dose as specified for a commercial ATSH 
enzyme preparation are followed. 

During the commercial demonstration test, Southwest Montana Alcohol also 
ran a conventional cook and fermentation to compare with the ATSH fermentation. 
Final ethanol concentrations and efficiencies were very similar. What was 
dramatic was the ferraenter turnover time for the conventional batch cook as 
contrasted to the ATSH process. The conventional batch cook and fermentation 
took a total of 84 hours to reach an ethanol concentration of 9.46 v/v%. The 
ATSH fermentation took a total of 58 hours to reach the same final ethanol con- 
centration. By using the ATSH process, the 26-hour savings could be turned into 
a 30% increase in ferraenter output without the cost of additional fermenters. 

ADM Test 



Two sets of tests were conducted in August and October 1986. Tests employed 
4 liter bench fermenters (New Brunswick Scientific). All used purified corn 
starch supplemented with corn steep liquor obtained from commercial production 
lines. A total of 20 separate fermentations were run, 8 during the August 
tests and 12 during the October tests. ATSH enzyme preparations from three 
different sets of experimental cultures were used. Variables tested included 
yeast strain, agitation, pH, starch concentration, enzyme dose, and temperature. 
Yeast strains included the distillery yeast (GB Fermentation Industries) used 
by RTI, the strain employed by ADM^and several experimental strains. Agitation 
was different for different fermenters because ferraenter agitation systems 
could not be operated intermittently. Some were not agitated; others were 
agitated continuously at low speeds. Starch concentration, dose rate and 
temperature are interelated variables affecting fermentation rate, conversion 

40 



efficiency and final alcohol concentration. Figure 18 shows results of dose 
rate and starch concentration experiments at optimum agitation using ADM 
yeast at 35°C ferraenter temperature. ATSH fermentations achieve conversion 
efficiencies and alcohol concentration equal to the conventional process. 
Fermentation rate depends on enzyme dose. Rates corapable to retention time in 
ADM continuous fermentation system were obtained at enzyme dose rates which 
appear to be economical compared with current costs for enzyme and cooking. 



41 



"<;>'iz ■■o<z ■•';::;'<£ 
,-.«siffi anL-^K isac!::; 




D. SYSTEM DESIGN 

A mass balance schematic, energy balance, process flow, and process flow 
schematic are included in Drawings BF503D6, BF503D7 and BF503D8. The assumptions 
for the mass and energy balances are presented in Appendix 4. 

The commercial plant would produce ATSH enzyme from mold grown on barley- 
based substrates using the following process. Barley from storage bins would 
be conveyed to a pearler/f laker that would produce pearling waste for livestock 
feed and barley flakes for mold culture substrate. The barley flakes would be 
mixed with chopped straw and nutrient supplements before being sterilized by 
steaming. The substrate would be cooled and augered into a clean reactor. 
Cooling would be within the augering system. The substrate would be inoculated 
with spores before entering the production reactor. Cycle time for each reactor 
would be four days, at which time the culture would be removed from the reactor 
and dried. The dried enzyme would be conveyed and ground before packaging into 
50-pound bags. The sacked enzyme would be stored and shipped as needed. 

Spores for inoculation would be produced in one of the reactors on a 
batch basis as needed. 

Equipment descriptions for the major process equipment, operating and 
control parameters, and production rates are described in the following text. 
IVhen applicable, equipment vendors and cost estimates are included with the 
equipment descriptions. 

Storage 

Barley storage requirements were based on three months of storatge. Volume 
requirements were calculated as follows: 

18,320,970 Ibs/yr barley x Bu/A8 lbs x 1.244 ft^/Bu x .25 yr = 
118,705 ft^ for 3 months of storage 

43 



Allowing one silo per month required 3 silos with individual volumes of 39,568 
ft3. Acron Manufacturing of Charlotte, North Carolina, sells a 42,750 ft^ 
corrugated metal silo. It is 33 feet in diameter and 50 feet high, complete 
with loading and unloading system, ladders, platforms, and aeration system. 
The price is approximately $16,000 per silo package. 

Barley straw for culture substrate would be purchased in bales and stored 
in stacks. The stacks could be covered but that probably is not necessary. 
Chopped straw could be stored in a concrete bunker. 

Pearler/Flaker 

Minnesota Grain Pearling estimated a package system cost at approximately 
$80,000. This system is rated at 2.5 ton/hour input. The package included 
all the necessary controls, equipment and utilities. A feeding system was not 
included. The discharge system consists of a discharge chute. 

A pearling/flaking operation operating 52 weeks/year, 5 days/week and 16 
hours/day would require a feed rate of 2.1 tons/hour. 

Straw Chopper 

A straw chopper processes the barley straw into pieces 1" to 3" in length. 
An 8-hour a day operation, 5 days/week, 52 weeks/year would require a 300 pound/ 
hour feed rate. The smallest choppers available are similar to International 
Harvesters Series 9000 tub' grinder. It has a 30 ton/hr feed rate based on 
alfalfa bales. The feed rate for straw would be approximately 15 tons/hr. 
This would require 21 hours of operation/year. Purchase cost for a stationary 
unit would be approximately $10,000. It should be possible to rent one for 3 
days during the year. 



44 



Mixing and Cooking 

Mixing and cooking can be combined into one operation. There are many 
types of equipment that could perform this combination efficiently. One that 
would be very satisfactory is made by Bepex Corporation. A Bepex Turusdisc 
Model TD36-12 has a hold-up volume of 77 ft^ and could process up to 6.25 
tons/hour on a continuous basis. A 5 day/week, 8 hr/day, 52 week/year opera- 
tion would require a 6.15 ton/hour feed rate. To load 4 reactors/week, a 10 
hour/day operation is necessary. A preliminary cost estimate for this model 
was $216,000. 

The Turusdisc is complete with temperature, time, rate, and speed controls. 
It was estimated that feed would enter at 65°F and exit at 210°F. Since the 
feed is 50% water, a specific heat of 1.0 Btu/pound °F was used. While this 
value is close, it may vary slightly. 



Cooling 



No specific equipment was specified for cooling. Residence time on the 
exit conveyor and reactor loading and start-up time should provide sufficient 
time for substrate cooling. Before plant construction, the dollar savings in 
heat recovery should be compared to the additional building and materials 
handling equipment. It may be desirable to install specific heat recovery 
equipment. 

Reactors 



The plant was sized with four reactors having a total volume of 24,880 
cubic feet or an individual volume of 6,220 cubic feet. Each reactor would be 
12 feet in diameter and 55 feet tall. Cycle time for one reactor would be 4 
days, including loading and unloading. A fifth day was allowed for cleaning 
and sterilization. Each reactor would be cycled once a week. 

45 



A reactor would be loaded with 123,000 pounds over a 10-hour period for 
a feed rate of 6.15 tons/hour. The discharge rate would be 6.02 tons/hour 
over 10 hours. 

The shell temperature would be controlled at 86°F while the bed tempera- 
ture would not exceed 100°F. Oxygen, carbon dioxide, inlet airflow, and exit 
gas flow would be monitored on a continuous basis. Airflows would be adjusted 
to keep exit oxygen and carbon dioxide within certain ranges. 

Dryers 

Many different types of dryers could be used ranging from a simple box 
dryer to vacuum dryers. Wysraont Company provides a rotating horizontal screen 
turbo dryer that would work very well. A turnkey packaged unit complete with 
feeders, wet scrubber, heating system, controls, instrumentation, and motors 
would cost $200,000. Wysmont claims heat recovery in the 74 to 78% range. 

Capacity of this unit is 15.5 tons/hour. Since each reactor's output is 
6.02 tons/hour, a smaller unit would work; however, this is the smallest 
standard unit Wysmont makes. If the plant were to operate 7 days/week rather 
than 5, the additional capacity would be used. 

Wet enzyme enters the dryer between 90 and 100°F. Tlie exit temperature 
would be approximately 75° F. 

Packaging 

St. Regis Company, based in Denver, Colorado, manufactures a packaging 
system used to package talc. The same system should work very well on dry 
enzyme. A complete turnkey system equipped with grinder and wet scrubber would 
cost $70,000. The final product would be packaged in 50 pound sacks. The 
system quoted would have an operating capacity of 4.5 to 5.1 tons/hour. The 



46 



required feed rate would vary from 2.25 tons/hour to 4.5 tons/hour depending 
on reactor scheduling. 

Conveyors 

It was estimated that a plant of this size would require approximately 
250 linear feet of converyors, including weigh belts. Conveyors and Equip- 
ment Company of Salt Lake City, Utah, suggested using $70/linear foot as an 
average cost figure for installed 24" wide, rubber-covered conveyor belt. 
Load cells and instrumentation for the weigh belts are included in the instru- 
mentation estimate. 

Boiler 

A boiler would be necessary to provide heat for flaking and cooking. 
Total heat requirement would be approximately 700 HP of steam. Velmco Sales of 
Missoula, Montana, estimated a Cleaver Brooks Boiler cost of $93,000. The 
complete system would produce 800 HP of steam at 30 psig and be fired with 
natural gas. 

Nutrient Pump and Tank 

A 10,000 gallon storage tank would be used for nutrient storage. Required 
pump capacity would be 12 gpm to match the feed rate to the mixer based on a 
10-hour operation. 

Inoculation 



Spores produced by one of the reactors in a batch operation on an as-needed 
basis would be slurried and stored in a holding tank. The slurry would be 
pumped and sprayed on the substrate before entering the reactor. The pump 
would have a capacity of approximately 1.5 gpm. 



47 



Miscellaneous 

Storage bunkers, hoppers, small tanks, etc. would depend on building 
location, shipping, operation schedules, and final equipment selection. 

Instrumentation 

One oxygen and one carbon dioxide monitor would measure concentrations for 
the exit gas of all four reactors. Concentrations of each component vary from 
to 21%. Individual flow and temperature controls are required. Airflow will 
require a proportional controller. Bed temperature will vary from ambient to 
100°F. 

Besides the instrumentation provided with the equipment and needed by the 
reactors, the following would be required: 6 level controls, 12 temperature 
controls, 19 flow controls, 5 pressure controls, and weigh belt instrumentation. 

James E. Rawley Company estimated the cost of this instrumentation at $93,000, 
including the programmable controllers. James E. Rawling Company is a vendor for ^^ 
Fisher Instruments. 

Building Requirements 

The primary requirement of a building is being able to wash the walls, 
ceiling and floors. Ideally, the grinding and bagging operation need to be 
separated from the rest of the process. This would facilitate dust collection. 

The building also needs an air conditioned storage area for bagged enzyme. 
The size of the storage area would be dependent on shipping and sales. A 12' x 
16' room should be adequate. 

The reactor room needs to have headroom above the reactors. This would 
require a ceiling height of approximately 65 feet. Tlie reactor floor space 
would be approximately 30' x 30' or 16' x 54' . 

The control room should be separate from the processing areas. (/^ 



IC would be desirable to locate the building and storage silo close to 
railroad tracks. This would make receiving large quantities of barley more 
economical and eliminate the need for truck scales. 

The building would require 440 volt, 3-phase power. Waste process heat 
could supplement building heat. 



49 



E. ECONOMICS 

Economics of ATSH enzyme production and use have been evaluated by market 
analysis and cash flow projections. 

1. Market Analysis 

The primary market for ATSH enzyme is the U.S. alcohol fuel industry. 
This area of market analysis will be discussed below. Other potential markets 
exist in overseas alcohol fuel production, food processing, alcoholic beverage 
production, and the textile industry. 

In 1986, U.S. alcohol fuel production capacity was about one billion 
annual gallons in about 200 plants. Production was about 750 million gallons. 
Over 90% of production was from corn and the remainder from other grains or 
starch crops. 

The industry consists of two market segments. A small number of large 
producers account for more than 80% of production. One company. Archer Daniels 
Midland Company (ADM), accounts for nearly 50% of production in four plants. 
Typical plant scale is 50 to 100 million gallons per year. Small producers, 
typically 10 million gallons per year or less, are the largest number of 
potential customers but less than 20% of the total market. 

The competitive advantage of ATSH enzyme results from reduced cost due to 
elimination of starch cooking. Cost savings include reduced process energy, 
cooling water, boiler operation, and maintenance, labor and capital equipment. 
Energy and capital cost savings are the most important. Cost savings vary 
depending on plant scale, cooking technology, local energy costs, and feedstock. 
RTI has evaluated savings for typical small plants based on Montana conditions 
and for large plants based on data from ADM and other companies. 



50 



# 



Small plants In Montana employ batch cooking and fermentation systems in 
which mash is heated, treated with enzymes, cooled, and fermented in the same 
tank. In these systems, direct energy requirements for cooling is in the range 
of 15,000 Btu/gallon of alcohol produced. Depending on fuel source and price, 
savings would range from 6 to 10|6 per gallon. Additional savings in cooling 
costs of about li per gallon would also be realized. In small plants, the 
simplicity of the one-step ATSH process would be an important advantage, pro- 
viding savings in labor and related costs. 

Analysis of large scale plants, particularly integrated, corn wet-milling 
plants, is more difficult because of the multiple process flows and the extensive 
use of heat recovery and recycle systems. Analyses in the technical literature 
vary widely depending on energy accounting assumptions. Direct energy con- 
sumption of typical jet cookers is about 9,000-13,000 BTU per gallon of alcohol 
produced. Assuming 50% heat recovery, net energy consumption would be 4,500 to 
6,500 BTU/gallon. Heat recovery is in the form of hot water which is generally 
in surplus and of low value in large wet milling operations. Costs more closely 
reflect direct energy consumption than the net figure after accounting for heat 
recovery. At $3.00 per million BTU cooking energy cost would be 2.7-3.9 cents 
per gallon, discounted perhaps 10-20% for heat recovery. In large, new plants, 
capital cost savings from elimination of cooking systems, boiler capacity, 
cooling system, and heat exchanger would be significant. 

Because of potential cost savings, the alcohol industry has been very 
interested in ATSH enzyme. Table 1 is a listing of alcohol fuel production 
companies which have contacted RTI regarding ATSH enzyme. These contacts were 
the result of very limited coverage of RTI in trade journals. These companies 
represent the majority of the U.S. industry. 



51 



Table 1. Alcohol Fuel Production Companies Which Have 
Contacted RTI Regarding ATSH Enzyme 



6 



.Company Name 


Location 


Annual Capacity 
(Gal 000,000) 


Alcotech 


Ring ling, MT 


7.5 


AE Montana 


Amsterdam, MT 


1.5 


SW Montana 


Dillon, MT 


.5 


Archer Daniels Midland 

Pekin Energy 

(CPC International) 


Decatur, IL 
Pekin, IL 


360 
(4 plants) 

68 


A. E. Staley Co. 


London, TN 


40 


Midwest Grain 


Atchison, KS 


14 


Dawn Enterprises 


Walhalla, ND 


14 


Tennol Inc. 




8 


Grain Processing Co. 


Muscatine, IW 


50 


New Energy of Indiana 


South Bend, IN 


52 


TCV Alcohol 


Greenburg, WI 


3 


CEPO Inc. 


Batavia, IL 


3 


Center Valley Alcohol 


Logan, WI 


2 


Greater Rockford Energy Corp. 


Rockford, IL 


2 


Agmart Inc. 


Monmouth, IL 


2 


Simplot 

TOTAL 


Boise, ID 


6 


627.5 



52 



Because of the small number of potential customers and their level of 
interest, marketing of ATSU enzyme will be almost entirely a function of cost 
savings and price. The ability to replace cooking processes will depend on 
marketing the ATSH enzyme for a price less than the combined cost of conventional 
enzymes and savings in energy and other process costs. Cash flow analysis 
described in the next section shows that ATSH enzyme can be produced and pro- 
fitably marketed at prices competitive with present commercial enzymes. 

SSC technology is not currently used in the U.S. for enzyme production. 
This technology needs to be proven at a small commercial scale to minimize 
technical, economic and market risks of very large production facilities needed 
to supply national markets. 

Initial plans for production of ATSH enzyme are based on regional markets 
composed of small scale producers. These markets can be supplied from a rela- 
tively small scale enzyme production plant which can serve as a prototype for 
larger facilities. Economics of enzyme price and cost savings are also more 
attractive in small scale plants, reducing economic risk in initial enzyme 
production. 

Regional markets consist of a number of plants in Montana, North and South 
Dakota, Idaho, Colorado, and Washington, with a combined capacity of about 30 
million gallons of production capacity. ATSH enzyme could also be supplied 
economically to small plants in the Midwest. Based on these markets, engineering 
considerations and capital cost estimates, preliminary plant designs and cash 
flow analyses are based on an ATSH enzyme plant designed to support 20 million 
gallons of annual alcohol production in the first year of operation. 



53 



2. Cash Flow Analysis 

RTI retained the accounting firm of Anderson ZurMuehlen to prepare cash 
flow analyses for ATSH enzyme production. Cash flow analyses and supporting 
engineering analyses are being used in ongoing efforts to raise private capital 
for construction of an ATSH enzyme production plant in Montana. 

A computer model was developed for the cash flow analysis. This allows 
rapid analysis of changes in model inputs resulting from technical progress, 
engineering changes, market considerations, and costs. Sensitivity to principal 
costs including capital, raw materials and finance assumptions can be readily 
evaluated. 

The cash flow analysis includes the following financial statements: 

Projected Balance Sheet 

Statement of Projected Results of Operation and Cash Flow - 5 Years 
Statement of Projected Results of Operation and Cash Flow - 1 Year 
Summary of Significant Projection Assumptions and Accounting Policies 
Supplementary Schedules 

The complete cash flow analysis is included as Appendix 5. The principal 
assumptions and inputs to the cash flow model are the capital cost estimates 
described in Section G and a computer model. Enzyme Production Analysis. 
Capital cost estimates are the basis for assumptions on finance requirements, 
debt and equity, assets, and depreciation. The enzyme production analysis 
provides the basis for extrapolating pilot plant data, market assumptions, 
revenue feedstock requirements, and other variable production costs. 

The enzjmie production analysis is shown in Table 2. This shows the 
functioning of the model and the actual inputs and calculated values used in 
the cash flow analysis. Inputs to the model are underlined. Other values are 
derived by calculation from these inputs. Important inputs are described by 
corresponding line numbers below. 



54 



i ; ENZYflE PRODL'CTIDN ANALYSIS prodanal.fsd 

- ' DATE: 2-!i-1987 Tiae: 16:53:50 

3 !♦ Note: L'nderlined areas are input; 

4 ! athers are calculated. Year 1 Year 2 Vgar ' 



6 ; PRODUCTION DATA 



■^ ■ 


!]o=e- ■; in: 3w/'.!: lar 


J.i\ 


3 ■ 


y.i?- En: Ibs.'gal Etoh 
Barley t-'bushel 


0.;75Z 

:.i5 


13 ; 
11 : 


Parley J/'.Gn 
rlalres \ of Barley 


137.53 
45.30 


i: ; 


Flakesiof trtai Hstkl 


3, '530 


i: : 


St'iH 'Gf total ^dstH 


3,3539 


14 : 


Pecoverv Rat? 


3.40 



3, -11 


3.3750 
2.33 


^91. 47 
65.39 


9.<?533 


3.3532 


3.40 



191.47 

d5.33 



3.63 



16 ; -FEEDSTDCK- 

17 ; Flakes-Cost S/ton J137.S2 till. 93 1141.03 1144.23 f~i " 

18 ; Straw-Cost t/ton 125.39 125.30 «25.30 

19 ! Flakes-Cost I/lb En: 10.128900 13.111333 10.111339 107173363 
28 I Straw-Cost I/lb En: 10.381039 10.391339 10.091039 i^'.Z^l 



3.2521 


J<?3.75 

65.33 


3.^523 


3.3533 


3.60 


1144.23 
125.30 



5,937 


1813,399 


625,909 


312 


17,812 


!, 500, 300 


6,259 


1326,122 


35.00 



35 : FEEDSTOCK 

36 ; flakes 4 straw 12,500,300 15,123,336 15,123,836 15.123,33 



' ^l3t^" ■' straw ton/yr 6,259 7,562 7,562 



39 1 By-product (Pearling) "; 35.00 35.09 

IB ;8v-product Ibs/yr 6,394,231 7,736,424 7,736^424 

41 iBy-product tons/yr 3,197 3,368 

^- '■ ''"trient Co st t/lb gn:yB 3.3913 9.3910 

'' ; Cheiical; I/lb en-vB 3.3003 0.8003 

44 ! Packaoirg l/lb en;yB 0.0003 9303 

45 I ' 

46 - SUfrARY. PRODUCTION DATA 

*7 ; En:Yge ?r:c? t/gal ETOH 13.120 ;b.i23 

43 :En:yije Price l/lb {9.329 10.329 

49 :SE7E'JJE, En:yse l/yr 12,399,983 12,903,762 

^3 : 3Y-product Price t/ton 143.30 ;4i.30 

51 IREVENUE, By-products 1127,335 1153,597 



3.3313 


3.3003 


0.9303 


10.124 


10.379 

13,440,257 

141.09 



3.3333 


3.3333 




19. 






19. 


455 


'4, 


128, 


329 




142 


:.30 



52 ;COSTS, Feedstock 1326,122 11,022,556 



53 ;COSTS, Total Other 112 



3, 


1 2' 


:64 


t; 


:- 


-<: 




b5, 


ixa 


3, 


,■=533 




,3533 




0. 


,60 



13.1:3363 
13.301339 

21 : Cost Fdstk l/lb En: 19.1098 10.1124 $3.1124 13 1149 "»-'i9 

22 1 -ANNUAL- ' '""" 
Ethanol Prod'')- aal/yr 19.9^9.399 24.193.315 27,499.333 Z2,llih,'^b3 -S ■:5'' 9-^3 



4,332 "=,374 
^.-537 i. 



24 ; En:yfle Ib/yr 7,533,330 9,374,302 ',374,392 

25 '■ En:yae ton/yr 3,753 4,537 4 5 

26 : Innoc'jiu.1 req'd !bs/yr 21.243 25,698 25.f:'g 2- .^'a -"^ top 
" ; TOTAL ENZYME Ibs/yr 7,521,248 9,190,308 9.130^338 ""TTisMM 7 •?g'igB 
23 1 TOTAL ENZYME tons/yr 3,761 4,550 4,553 4,553 '""4,"558 
29 : Flakes- Input Ibs/yr 11,375,000 14,367,644 14,367,644 14,367!644 \i,''t,1%^ 
38 ; Flakes- Input tons/yr 5,937 7,134 7,134 "" 7'io4 ' ''TI?4 
311 Flakes - Cost l/yr 1813,309 11,013,103 11.913,183 II.036,'l28 H 0'6'r3 
32 I Straw- Input Ibs/yr 625,909 756,192 756,192 756!l92 "754."l9'' 
33; Straw- Input tons/yr 312 378 378 -73 ''373 
34 ; Straw - Cost l/yr |7,812 $9,452 19,452 |9.452 19 4^ 



562 7.562 



33 ; Cost-flakes+straw l/yr 1826,122 11,022,556 11,022,556 11.345,531 ll.g 



33 35.08 35.38 

736,424 7,736,424 

3,863 

3.3313 



:■ 


.363 




3313 




333^ 




333" 






13, 


,563 


15,333, 


,"=! 


142.39 



»I53,597 llo2,465 

11,322,556 11,045,531 11,345.581 



*1*.51' 114,519 114,519 114,519 



Table 2. 
55 



Line 7 - Dose: This is the dose rate expressed as weight percent of 
enzyme in mash. This value is based on results from pilot plant 
tests. Dose rate is forecast to improve with continued technical 
progress. 

Line 9 - Barley: Price of barley in dollars per bushel. The value is 
the 7-year average price in Montana. 

Line 11 - Flakes % of Barley: The percentage of barley recovered for use 
as mold culture substrate. 

Line 12 - Flakes % of Total Feedstock: The proportion of flakes used in 
the total substrate. 

Line 13 - Straw: As portion of other components added to processed barley 
for physical bed structure. 

Line 14 - Recovery Rate: This is the amount of enzyme recovered from 
culture substrate. This value is from pilot plant results. 

Line 18 - Straw: Cost in dollars per ton. 

Line 23 - Ethanol Production Gallons Per Year: The estimated market 
for ATSH enzyme. 

Line 26 - Inoculum Required: The amount of culture required for spore 
preparations. 

Lines 27-38: Calculated values for annual enzyme production requirements, 
production inputs and costs based on dose rate, barley costs and 
market size. 

Lines 39-41: These are calculated values for by-product recovery from 
processing inputs (Line 11) and feedstock inputs (line 29). 

Lines 42-44: These are inputs for nutrients used in culture substrates, 
chemicals used in various process steps and packaging costs. 

Line 47 - Enzyme Price: This is market price of enzyme to alcohol pro- 
ducers. The input is expressed as enzyme price per gallon of alcohol 
for ready comparison to conventional enzymes. The value of $.12/ 
gallon is based on current enzyme prices and cost savings at typical 
small alcohol plants. 

Line 49 - Revenue: Revenue is calculated from price (line 47) and market 
inputs (line 23). 

Line 50: By-Product Price: This is an input value based on market price 
for barley processing by-products. The value is based on sale as 
cattle feed. 

Line 51: By-Product Revenue: By-product revenue is calculated from by- 
product amounts and price. 

56 



Input values reflect current levels of technical progress and market 
conditions. Tlie cash flow generated from these values together with capital 
cost estimates and finance assumptions show that ATSH enzyme productioa will be 
profitable at the projected scale of production. This is shown in the 5-Year 
Statement of Projected Results of Operation and Cash Flow, Table 3. The cash 
flow assumes sales increases (in terns of alcohol production) from 20 million 
gallons per year to 38 million gallons per year in year 5. This reflects full 
utilization of the initial plant capacity. Net after tax income increases 
from $68,138 in period 1 operation to $1.9 million in period 5. Corresponding 
cash balances are $484,002 in period 1, increasing to nearly $4.8 million in 
period 5. 

Contractually required repayment of grant funds disbursed by DNRC for 
these projects is not shown in the cash flow analysis pending a determination 
by tax counsel of the cost category for this obligation. Repayment, however, 
is a recognized cost which would affect cash available at the end of each 
year. 

Cash flow analysis shows that a small scale ATSH enzyme production plant 
can be profitable, providing both a demonstration of the technical and commer- 
cial potential of SSC and generating sufficient income to support expansion of 
ATSH enzyme production to serve national markets. 



57 



F. GRANT ADMINISTRATION 



1. Work Schedule 



For Che first 14 months of this project, from May 1984 through June of 
1985, all work and project milestones for Grant Agreement RAE-84-1044 were 
completed by their proposed deadlines. In July of 1985 work was begun on two 
additional projects, both of which were related to and impacted the scope of 
RAE-84-1044. 

Grant Agreement RAE-85-1055 from DNRC was awarded in June of 1985 and 
work commenced in July. This project, for work on feedstock processing for 
culture substrate preparation and improved process monitoring and control, 
supplemented the work being conducted in RAE-84-1044. As a result of discus- 
sions with DNRC staff, it was decided to run the two projects concurrently 
and submit one final report on the research from both projects. Ammendraent 
No. 3 to RAE-84-1044 extended the completion date of RAE-84-1044 from January 
1986 to May of 1987. 

Also in June of 1985 RTI obtained a Small Business Innovation and Research 
Program, Phase II award from the U.S. Department of Energy for related research. 
This award more than doubled the amount of funds available for advancement of 
the solid state culture technology. 

In order to maximize the effectiveness of the research and development 
being conducted and for best utilization of combined funds from DNRC and U.S. 
D.O.E. the three programs were integrated wherever possible. The combined 
effect of this integration was to extend the performance period of both 
RAE-84-1044 and RAE-85-1055. Figures 19 and 20 show the timelines for both 
of the DNRC projects. 



59 



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A technical delay in RAE-84-1044 occurred in Task 6, Milestone 5, Commer- 
cial Demonstration. The commercial demonstration was delayed 8 months while 
the different pilot plant reactor designs and operating parameters were tested 
and a sufficient quantity of enzyme was produced to conduct the commercial 
demonstration test. 

Some technical delays occurred in RAE-85-1055 (see Figure 5) which can be 
attributed to long lead times in equipment delivery and the complexity in soft- 
ware development as part of tasks in Component II, Improved Monitoring and 
Control. 

The administrative and technical delays in the two projects are probably 
not unique to research projects. What is unique in the projects is the inte- 
gration of ATSH enzymes and cellulase to make solid state culture systems 
technology marketable to potential private investors. 

2. Budget 

The contract budgets and costs billed to the contract by budget category 
for RAE-84-1044 and RAE-85-1055 are shown in Tables 4 and 5, respectively. 
The billed costs for RAE-84-1044 match the contract budget except for $7008 in 
equipment money that was moved to contracted services. In RAE-85-1055, it 
should be noted that $8160 was authorized by DNRC (letter dated December 29, 
1986) to be moved from contracted services to salary (salary, fringe benefits, 
laboratory indirect, and indirect). 

From May 1984 to May 1987, RTI will have expended research funds of 
$931,551 on solid state culture development for enzyme production (see Table 
6). The two enzymes resulting from this research are ATSH enzymes funded by 
DNRC and cellulase funded by the U.S. DOE. While the research was conducted 
on different enzymes, the results can be used to complement each other. DNRC 

62 



research provided a culture system and experimental approach that was adapted 
and built on for the cellulase research. 



Tabl 


e 4. Budget for Grant 


Agreement No. RAE- 


-84-1044 






Contract 


Billed 






Budget 


Costs 


a. 


Salary 


$118,370 


$118,370 


b. 


Fringe Benefits 


39,062 


39,062 


c. 


Contracted Services 


6,000 


13,008 


d. 


Supplies and Materials 


13,302 


13,302 


e. 


Communications 








f. 


Travel 


926 


861 


g. 


Lab Indirect 


24,859 


24,859 


h. 


Equipment 


28,955 


21,947 


i. 


Indirect 


53,526 


53,526 




TOTAL 


$285,000 


$284,935 



63 



Tabl 


e 5. Budget for Grant 


Agreement No. RAE- 


-85-1055 






Contract 


Billed 






Budget 


Costs 


a. 


Salary 


$ 34,741 


$ 34,741 


b. 


Fringe Benefits 


11,464 


11,464 


c. 


Contracted Services 


19,040 


19,040 


d. 


Supplies and Materials 


6,385 


6,385 


e. 


Communications 


350 


128 


f. 


Travel 


5,000 


1,483 


g- 


Lab Indirect 


7,473 


7,473 


h. 


Equipment 


19,955 


19,955 


i. 


Indirect 


29,065 


29,065 




TOTAL 


$133,473 


$129,734 



Table 6. List of RTI Biofuel Contracts for Solid State Culture Development | 


RTI 

Control 

No. 


Contract 
No. 


Agency 


Enzyme 


Title 


Amount 


BF401 


RAE-84-1044 


DNRC 


ATSH 


ATSH Enzjmie Commercial 
Development & Research 


$284,935 


BF503 


RAE-85-1055 


DNRC 


ATSH 


ATSH Commercial Develop- 
ment: Improved Barley 
Processing & Culture 
Monitoring & Control 


$129,734 


BF403 


DC-AC03- 
84ER80188 


DOE 


Cellulase 


SBIR Phase I - Solid State 
Culture of Trichoderma 
reesei for Cellulase 


$ 49,566 


Production 


BF503 


DE-AC03- 
84ER80188 


DOE 


Cellulase 


SBIR Phase II - Solid State 
Culture of Trichoderma 
reesei for Cellulase 
Production 


$467,316 


TOTAL 


$931,551 



64 



C. RESULTS AND CONCLUSIONS 

1. Major Findings and Recommendations 

The two projects resulted in successful pilot scale development of ATSH 
enzyme production technology. Preliminary design and equipment specifications 
for a commercial plant to produce enzyme for a regional market of 20-50 million 
gallons of annual alcohol fuel productions were developed. The plant could be 
built in Butte, Montana for an estimated $1.1 million. In full production 
(year 5 of operation), the plant would employ about 60 people. Economic analysis 
shows an attractive return on investment at this scale of operation. 

ATSH enzyme has beem commercially demonstrated for both small and large 
scale alcohol production. Elimination of cooking will result in significant 
operating cost savings at any scale of alcohol production. Capitol cost 
savings of up to 30% of total conventional plant cost could also result from 
elimination of cooking. 

Because of potential cost savings, the alcohol industry has expressed 
considerable interest in ATSH enzyme. RTI has been contacted by firms 
representing over 90% of U.S. alcohol fuel production. This level of interest 
Indicates a national as well as regional market. Successful operation of a 
small commercial facility will provide the technical and financial base to 
support expansion to supply national markets. 

Based on technical, economic and market analysis, RTI initiated work to 
raise private capital to construct and operate a commercial ATSH enzyme pro- 
duction plant to serve regional alcohol fuel markets. 

Solid state culture systems developed for ATSH enzyme production can be 
used to produce a number of mold derived products. As a result of the exper- 
tise developed through DNRC funding, RTI has obtained over $600,000 in federal 

65 



grants for work on three additional 

solid state culture products. These are cellulase, ligninase and a mold 
based biopesticide. All of the products have large potential markets and can 
be produced in Montana using agricultural crops or waste as the principal 
feedstocks for plant operation. RTI has completed a business plan and is 
working to raise private capital to support additional development work and 
commercial production of these additional products. 

2. Permits, Licenses, and Authorizing Agencies 

A number of permits and licenses are necessary to build an ATSH enzyme 
production facility. The permits and licenses can be divided into two cate- 
gories: 1) Permits necessary to starting any business and 2) permits specific 
to an ATSH enzyme plant. Permits that fall under category one are such things 
as business licenses and building permits. Special permits deal more with 
environmental and operating permits that are more directly related to the type 
of business. The two categories of permits are discussed below. The inform- 
ation on permitting and licenses was taken from DNRC's publication "Montana's 
Bioenergy Project Permitting Guidebook," July 1986. 

a. General Business Licenses and Construction Permits 

RTI plans to start a new company called Mycotech, Inc. to produce ATSH 
enzymes. The business must go to the Secretary of State and file articles of 
incorporation, and register a business name and trademark. Other business 
responsibilities include obtaining a local business license, obtaining a federal 
tax identification number, registering with the Montana Department of Revenue 
as an employer for income tax purposes, filing a state withholding tax regis- 
tration, registering for unemployment insurance and obtaining worker's compen- 
sation insurance. 

66 



RTI plans to build the ATSH enzyme plant In the Butte, Silver Bow area if 
at all possible. The local planning board would have to approve any proposed 
site for zoning conpliance. Before construction can begin the local building 
department will be contacted to get the necessary building, plumbing, electrical 
and mechanical permits. The local fire department will be contacted to make 
sure the building meets fire escape, fire alarm and fire extinguisher require- 
ments, and complies with the Uniform Fire Code. 

b. Permits and Licenses for the ATSH Enzyme Plant 

The state Department of Agriculture requires several permits for businesses 
buying agricultural commodities and selling commercial feed. A Commodity 
Dealers License is required for any business involved in the buying of Montana 
agricultural commodities. A Warehouseman's License is required to store the 
grain. A license fee based on the volume of grain handled per year plus a 
minumum bond of $20,000 is required for each license. Tlie Department also has 
specific business requirements and requires detailed reporting and record 
keeping. Selling the by-products of the pearling process will require a Feed 
Marketing Permit from the state agriculture department. All commercial feeds 
must comply with labeling format requirements, brand and product name specifi- 
cations, expression of guarantee requirements, statements of ingredients and 
directions for use and other requirements as defined by the laws and rules. 

Environmental permits are required for emissions or effluents that affect 
air quality or water quality or are solid or hazardous waste. RTI anticipates 
that no environmental permits will be necessary for the ATSH enzyme production 
plant. The only gaseous emissions that are predicted from the production 
facility are CO2 from the natural gas fired boiler and water vapor from the 
drying process. Grain processing, enzyme grinding and bagging will generate 

67 



particulates that will be controlled with wet scrubber technology integrated 
with the appropriate processing equipment. It is planned to locate the pro- 
duction facility in an industrial zone that is connected to the Butte, Silver 
Bow sewage treatment facility. The liquid effluents predicted are waste 
scrubber water, cooling water and rinse water that is used to clean reactors 
and other processing equipment. Solid waste from the facility will include 
floor sweepings and out-of-spec enzyme. None of this material is classified 
as hazardous waste. The solid waste material that cannot be recycled or sold 
as a by-product will be disposed of in the Butte-Silver Bow landfill. 

The ATSH enzyme plant will require several other operating licenses. The 
boiler must be licensed by the Department of Labor and Industry (DOLI). The 
persons operating the boiler must have the proper grade of Boiler Operator's 
License as specified by DOLI. The enzyme plant will fall under the federal 
Occupational Safety and Health Administration (OSHA). OSHA does not require 
a license or permit, but does require compliance with all federal regulations. 
The only potential problem with the enzyme plant is spores and dust in the work 
environment. All process and materials handling equipment will be designed to 
minimize dusting problems and integrated with dust control equipment. 

3. Additional Development 

Solid state culture is not currently used in the U.S. for enzyme produc- 
tion. Development plans include a period to design and test one commercial 
scale culture reactor and associated processing equipment. Commercial pro- 
duction would follow by replication of a tested commercial scale reactor. 
This approach minimizes scale-up risk. 

Experimental work would continue during design construction and testing 
of the commercial plant. Additional work includes experiments to better define 

68 



patentable aspects of the technology, develop a rapid assay procedure for 
quality control and continued work to improve enzyme production efficiency. 
Enzyme production economics is very sensitive to enzyme dose rate. As part 
of the final development work, RTI plans a continuing effort to select improved 
strains, identify rate limiting steps in raw starch hydrolysis and better 
define metabolic control of enzyme production. This work would continue during 
commercial operation. 



69 



APPENDIX 1 

Computer Software Monitoring 
and Control System 



DAS So-ftware Listing for ' DEL^.'ER2- BAS 

13 REM THIS PROGRAM IS DESIGNED TO OPERATE VALVES AND READ ANALYZERS 

20 REM AND THERMOCOUPLES TO AUTOMATE THE OPERATION AND DATA ACQUISITION 

30 REM FOR THE RTI CULTURIN6 SYSTEM. 

40 REM WRITTEN BV STEVE LUNDBERG 3/86 

50 REM MODIFIED: 
i0 REM 
^0 REM 
30 REM 

90 REM THE PROGRAM HAS FOUR PRINCIPAL PARTS: 

100 REM 1. PARAMETER SETUP LINES 1000-3999 

110 REM 2. DATA ACQUISITION SYSTEM SETUP LINES 4000-4999 

120 REM 3. OPERATION LINES 5000-7999 

130 REM 4. SUBROUTINES LINES 8000-9999 
140 REM 
150 REM 

160 REM PARAMETER SETUP: 1000-39q'P 

170 REM IOCONFIG.DAT: RANDOM FILE CONTAINING THE NAMES OF ALL 

ISO REM DEVICES ATTACHED TO ALL KIETHLEY I/O CHANf- 

190 REM FIRSTREACTOR, LASTREACTOR: MUST BE UNBROKEN SEQUENCE 

200 REM SV.$(I<,I): STRING ARRAY CONTAINING NAMES OF VALVES FOR 

210 REM INLET AND OUTLET OF EACH REACTOR 

220 REM TC*(I): STRING ARRAY CONTAINING NAMES OF THERMOCOUPLES 

230 REM FOR EACH REACTOR 

240 REM H2DWAIT7.: NUMBER OF SECONDS TO WAIT FOR STABILIZATION 

250 REM BEFORE READING HUMIDITY METER 

260 REM C02WAIT-/.: NUMBER OF SECONDS TO WAIT FOR STABILIZATION 

270 REM BEFORE READING C02 METER 

280 REM 02WAIT7.: NUMBER OF SECONDS TO WAIT FOR STABILIZATION 

290 REM BEFORE READING 02 METER 

300 REM 02STRTLISTf: LIST OF VALVES TO BE OPENED AT BEGINNING OF 

310 REM 02 MEASUREMENT CYCLE 

320 REM 02STRTPT2i:: LIST OF 3 VALVES TO BE CLOSED AFTER PTl 

330 REM STABILIZES 

340 REM 02READY:f: LIST OF 2 VALVES TO BE OPENED PRIOR TO 

350 REM READING 02 

360 REM 02END:|:: LIST OF 3 VALVES TO BE CLOSED AT END OF 02 R 
370 REM 
380 REM 

390 REM SOFT5O0 SYSTEM SETUP: 4000-4999 

400 REM INITIALIZE SOFT500: CALL INIT 

^10 REfl INITIALIZE VARIABLES: SET VALUE, VSTAT , HRX , MIN7., SEC7., 

^20 REM DATE7., MOV., YR"/. ALL = 

^30 REM SET CURDEP = 1 

^^0 REM 0PEN5?<: SOFT500 BIT ARRAY < 1 , 1 , 1 , 1 , 1 ) 

450 REM CL0SE3?<: SOFT500 BIT ARRAY (0,0,0) 

460 REM 0PEN2?': SOFT50O BIT ARRAY (1,1) 

470 REM MAKE lONAMES FOR SOFT500 

430 REM FOR EVERY CHANNEL ON EACH CARD ASSIGN lONAME 

490 REM FOUND IN IOCONFIG.DAT 

500 REM NUMTCy.(I): INTEGER ARRAY CONTAINING NUMBER OF TC ' S IN 

510 REM EACH REACTOR 

520 REM CALCULATE TOTAL # OF SAMPLES 

530 REM SET UP MAIN ARRAY FOR STORAGE OF EXPERIMENT VALUES 
540 REM 
550 REM 

'560 REM OPERATION: 5000-7999 



PAGE 2 

570 REM ASK IF READY TO BEGIN 

530 REM MAIN DATA LOOP 

590 REM LOOP OVER ALL REACTORS IN USE FOR BOTH INLET AND OUTLET 

600 REM TIMESTAMP ^ 

610 REM READ TC ' S 

620 REM READ PT0 

630 REM READ GAS TEMP 

640 REM READ HUMIDITY 

650 REM READ C02 % 

660 REM READ 02 '/. 

670 REM PICK UP ADJUSTING INFORMATION 

6S0 REM DISPLAY SELECTED DATA 

690 REM MAKE CONTROL CALCULATIONS 

700 REM SET UP CONTROL LOOP FOR 02X , C02/: 

710 REM END LOOP 

720 REM EXPERIMENT CLEANUP (PURGE, CLOSE ALL '-v-ALVES , SHUT DOWN SYSTEM) 

730 REM STORE MAIN ARRAY ON DISK WITH PROPER NAME 

740 REM END 

750 REM 

760 REM 

1000 REM ***i<-* PARAMETER SETUP ***** 

1010 DIM T I TLE*: ( 2 > , CHANNAME* (16) , 3V:f (2,10), TC:t ( 1 ) , NUN rCX < 1 O ) , MhASf (17^ 

1020 REM SECTION TO SET UP REACTOR #S AND C02 AND 02 LEVELS AND TIMES 

1 030 OPEN " I" , #2 , " P ARAM . DAT " 

1 040 I NPUT #2 , TEMP J-- , F I RSTREACT/: , TEMP F , LASTRE ACT X 

1050 INPUT #2,TEMP$,C02LEyELl"/.,TEMP$,C02LE'v'EL2/:,TEMP:f ,C02TIME1"': ' ,C02TIMb2/: 

1 060 I NPUT #2 , TEMP* , 02LEVEL 1 7. , TEMP-Jp , 02LEVEL27. , TEMP* ,, 02T I ME 1 "•;. ' , 02T I ME2/;. 

1070 CLOSE #2' 

1080 C02TIME27. ^ C02TIME1/: 

1090 02TIME2y. = 02TIME1/: ^^ 

1100 CLS 

1105 COLOR 2 

1110 PRINT SPC(10) "REACTORS IN USE" 

1120 PRINT 

1130 PRINT "NOTE: REACTORS IN USE MUST BE IN A CONTINUOUS SEQUENCE,." 

1140 PRINT 

1150 PRINT "FIRST REACTOR: " FIRSTREACTV. " LAST REACTOR: " LASTREhCT/I 

1 1 60 PR I NT 

1170 INPUT "ARE THESE ASSIGNMENTS OK? [Y3",RESP* 

11S0 IF (RESP* <> "n") AND (RESP* <> "N") GOTO 2110 

1190 LOCATE 10 

1200 INPUT "ENTER NUMBER OF FIRST REACTOR: " , FIRSTREACTV. 

1210 IF (FIRSTREACTV. > 10) OR (FIRSTREACTV. < 1) GOTO ir?0 

1220 LOCATE 12 

1230 INPUT "ENTER NUMBER OF LAST REACTOR: " , LASTREACT--^. 

1240 GOTO 1100 

2110 REM PARAMETER SETUP 

2115 COLOR 2 

2120 REM ARRAY SETUP FOR SVt(K,I) 

213.0 OPEN " I" ,#2, "SVARRAY.DAT" 

2140 FOR K = 1 TO STEP -1 

2150 I NPUT #2 , SV* ( K , 1 ) , SV* ( K , 2 ) , SV* ( K , 3 ) , SV* ( K , 4 ) , SV* ( K , 5 ) , SV* ( K , 6 ) , SV* ( K , 7 ) , J 

* ( K , a ) , SV* ( K , 9 ) , SV* ( K , 1 ) 

2160 NEXT K 

2170 CLOSE #2 A 



PAGE 3 

2180 CLS 

2190 LOCATE 7 

2200 PRINT "REACTOR INLET/OUTLET VALVE rJANEB" 

2213 PRIMT 

2220 PRINT "REACTOR i 2 3 4 5 6 7 8 9 1 

2230 PRINT 

2240 PRINT "INLET ", 

2250 PR INT US I NG " \ \ " ; SV$ (1,1); SV* (1,2); S'-.'S (1,3); SV;*: (1,4'; SV* ( 1 , 5 ) ; SV ; 

V* ( 1 , 7 ) ; SV-f (1,8); S Vf (1,9); 5\>t- ( 1 , 1 ) 

2260 PRINT 

2270 PRINT "OUTLET ", 

2280 PRINT USING "\ \ " ; SV* (0 , 1 ) ; SV# (0 ,, 2) ; SV-^ ( , 3) ; SVJ? (0 , 4) ; SV:ir .;0 , 5) ; SV 

V* ( , 7 ) ; SV* (0,8): SV* (0,9); S\-'T (0,10- 

2290 LOCATE 20 

2300 INPUT "ARE THESE ASSIGNMENTS OK CY]",RESP^ 

2310 IF ''RESRif <:•■ "N") AND (RESP* <> "n") GOTO 2620 

2320 LOCATE 22 

2330 PRINT "ENTER VALVE POSITION, VALVE NAME" 

2340 PRINT "EXAMPLE: I2,SV1B FOR INLET TO REACTOR #2." 

2350 INPUT SPOT-t, VALVE f: 

2360 FIRSTLET^ = MID* (SPOT t , 1 , 1 ) 

2370 IF (FIRSTLETJ: = "I") THEN K = 1 

2380 IF (FIRSTLET-* = "0") THEN K = 

2390 IF vFIRSTLET:f <> "I") AND (FIRSTLET* <> "O") GOTO 2580 

2400 3EC0NDLET* = MID-f (SPOT:|: , 2 , 2) 

2410 I = VAL(SECONDLET*) 

2420 IF (I < 1) OR (I > 10) GOTO 2590 

2430 TEMP* = MID* (VALVE*, 1 ,2) 

2440 IF TEMP* <> "SV" GOTO 2590 

2450 IF LEN( VALVE*) > 4 GOTO 2590 

2460 SV*(K,I) = VALVE* 

2470 INPUT "CHANGE ANOTHER-^ CN]",RESP* 

2430 IF (RESP* = "Y") OR (RESP* = "y") GOTO 2320 

2490 OPEN "0" ,#2, "SV ARRAY. DAT" 

2500 FOR K = 1 TO STEP -1 

2510 FOR N = 1 TO 9 

2520 PRINT #2,SV*(K,N) ",", 

2530 NEXT N 

2540 PRINT #2,SV*(K,10) 'NO COMMA AT END OF RECORD 

2550 NEXT K 

2560 CLOSE #2 

2570 GOTO 2620 

2580 PRINT "ERROR' USE I FOR INPUT SIDE, OUTPUT SIDE," 

2590 PRINT "NUMBER IN RANGE 1-10 FOR REACTOR, 4 CHAR. NAME STARTING W/ SV 

2600 GOTO 2320 

2610 REM END OF SV* ARRAY UPDATE 

2620 REM SET UP TC* ARRAY 

2630 OPEN "I" ,#2, "TCARRAY.DAT" 

2640 FOR I = 1 TO 10 

2650 INPUT »2, TCr ( I) 

2660 NEXT I 

2670 CLOSE »2 

2680 CLS 

2690 LOCATE 5 

2700 PRINT "REACTOR TC NAMES 

2710 PRINT 



PAGE ^ 

2720 FOR I - 1 TO 1(3 

2730 PRINT " " I TAB (10) TC:J: ( I ) 

2740 NEXT I 

2750 INPUT "APE THESE ASSIGNMENTS OK-^^ Cy:",PESP-t ^' 

2760 IF (RESP-:F <> "N") AND (RESP:? v; "n") GOTO 2930 'ALL OK 

2770 LOCATE 20 

277 1 PR I NT SPACE* ( 70 ) ; PR I NT SP ACE-t- ( 70 ) : PR I NT SPACEJ^ ( 70 ) ?. PR I NT SPACE* ( 70 ) 

2772 LOCATE 20 

2780 INPUT "ENTER REACTOR NUMBER: ",I 

2790 IF a < 1) OR (I > 10) GOTO 29Q0 'ERROR ROUTINE 

2800 PRINT "ENTER LIST OF TC ' S SEPARATED BY SINGLE SPACES." 

2810 INPUT TC*(I) 

2840 OPEN "0" ,#2, "TCARRAY.DAT" 

2850 FOR I = 1 TO 10 

2860 PRINT #2, TC* ( I ) 

2870 NEXT I 

2830 CLOSE #2 

2890 GOTO 2680 

2900 PRINT "REACTOR # MUST BE BETWEEN 1 AND 10 I NCLUSP-'E „ " 

2910 GOTO 2770 

2920 REM END OF TC* ARRAY UPDATE 

2930 REM SET UP WAIT TIME 

2940 OPEN "I" ,#2, "WAIT.DAT" 

2950 INPUT #2,TEMP*,H20WAIT7. 

2960 I NPUT #2 , TEMP* , C02WA I T/1 

2970 INPUT #2,TEMP*,02WAIT7. 

2980 CLOSE #2 

2990 CL3 

3000 LOCATE 5 ^ 

3010 PRINT "ANALYZER STABILIZATION DELAYS" ^ 

3020 PRINT "1) H2n","2) C02","3) 02" 

3030 PRINT TAB (4) H2DWAIT-/. TAB (13) C02WAIT7. TAB (32) C2WAIT7, " SECS" 

3040 LOCATE 10 

3050 INPUT "ARE THESE ALL OK'^ CY: " , RESP* 

3060 IF (RESP* <:> "N") AND (RESP* <> "n") GOTO 3190 

3070 INPUT "ENTER # TO CHANGE: " ,NV. 

3030 ON NV. GOTO 3100,3130,3160 

3090 GOTO 2990 

3100 LOCATE 15 

3110 INPUT "ENTER NUMBER OF SECONDS TO WAIT FDR HUMIDIT\' METER STABILIZATION: 

H20WA I TV. 

3120 GOTO 2990 

3130 LOCATE 15 

3140 INPUT "ENTER NUMBER OF SECONDS TO WAIT FOR C02 METER STABILIZATION: " , CO: 

I TV. 

3150 GOTO 2990 

3160 LOCATE 15 

3170 INPUT "ENTER NUMBER OF SECONDS TO WAIT FOR 02 METER STABILIZATION: " , 02W(- 

7. 

318.0 GOTO 2990 

3190 OPEN "0" ,#2, "WAIT.DAT" 

3200 PRINT #2, "H20, " ,H20WAIT7. 

3210 PRINT #2,"C02, " ,C02WAITy. 

3220 PRINT #2, "02," ,02WAIT7. 

3230 CLOSE #2 |^ 

3240 REM THE FOLLOWING ARE LISTS OF VALVES USED IN 02 MEASUREMENT ~ 



PAGE 5 

3250 REM THESE LISTS CORRESPOND TO SETS OF VALVES THAT ARE 



MEASUREMENT 



'CLE 



4 AS IOCHAN0*, 



4 AS lOCHANl*, 

4 AS 



AS lOCHANUl 
A3 lOCHANl' 



3260 REM ACTUATED SIMULTANEOUSLY DURING 

3270 02STRTLIST* = "SV4 SV5 SV6 SV7 SV9" 

3230 n2STRTPT2-.f = "SV4 SV6 SV7" 

3290 02R£HDY-r = "SV6 SV7" 

3300 02END:J: = "SV6 SV7 SV9" 

3310 REM SET UP HARDWARE CHANNEL INFO 

3320 OPEN "R" ,ttl , "IOrONFIG.DAT" ,72 

3330 HALF7. = 1 

3340 FIELD #1 , 4 AS IOCARDjF, 4 AS lOSLOT-t- 

CHAN2f, 4 AS I0CHAN3:|:, 4 AS I0CHAN4$, 4 A3 lOCHANS:*:, 4 AS I0CHAN6*, 

:t:,32 AS LASTS* 

3350 FIELD #1 ,40 AS FIRST10-f, 4 AS lOCHANS*, 4 AS lOCHAN^-.l:, 4 

S lOCHANllf, 4 AS I0CHAN12*, 4 AS I0CHAN13*, 4 AS I0CHAN14*, 4 

3360 REM HALF OF CHANNAMES 

3370 CLS 

33S0 TITLE* (1) = "CARD SLOT CH-0 CH-1 CH-2 CH-3 CH-'^ CH-5 CH-6 CM 

ZZ-^Q TITLE* (2) = "CARD SLOT CH-8 CH-9 OHIO CHll CHI 2 CM 13 CHI 4 CH 

3400 REM DISPLAY CONFIGURATION OF S CHANNELS ON ALL 10 SLOTS 

3410 PRINT TITLE* (HALF-/.) 

3420 FOR I = 1 TO 10 

3430 GET 1 , I 

3440 LOCATE id - 1) *• 2 ^ + 3 

3450 ON HALF/l GOTO 3460,3430 

3460 PRINT IOCARD* IOSLOT* IOCHANQ* lOCHANl* I0CHAN2* I0CHAN3* inCHAN4* iq 

I0CHAN6* I0CHAN7* 
3470 GOTO 3490 

3480 PRINT IOCARD* IOSLOT* I0CHAN3* I0CHAN9* lOCHANlO* lOCHANl 1* inCHHN12:^ 
N13* lOCHANl 4* lOCHANl 5* 
3490 NEXT I 

3500 REM CHECK FOR CHANGES 
3510 LOCATE 23 

3520 INPUT "ARE THESE SLOT/CHANNEL ASSIGNMENTS OK? CY:",RESP* 
3530 IF (RESP* "n") AND (RESP* <> "N") GOTO 3630 
3540 LOCATE 23 

3550 INPUT "ENTER SLOT , CHANNEL (FORMAT: 3,7) OR 0,0 
3560 IF (SL0T7. = 0) AND (CHAN"/. = 0) GOTO 3670 
3570 IF (SL0T7. > 10) OR (SLOT'/. < 1) GOTO 3540 
3530 IF (CHANV. > (7 + (3 * (HALF7. - 1)))) OR fCHANV. 



TO EXIT: " ,SLOT/:,CHe 



(HALF7. -1) ) ) GC 



: EXAMPLES TE0A,SV1A) " , lOCHAN* 





3590 LOCATE 23 

3600 INPUT "ENTER NAME OF INSTRUMENT. 

3610 IF LENdOCHAN*) <= 4 GOTO 3650 

3620 LOCATE 23 

3630 PRINT "NAME CANNOT BE MORE THAN 4 CHARACTERS LONG."; 

3640 GOTO 3590 

3650 GOSUB 9000 'PUT NAME IN IOCONFIG.DAT FILE 

3660 GOTO 3540 

3670 REM END OF IOCONFIG.DAT HALF ADJUSTMENTS 

3630 IF HALF'/. = 2 GOTO 3710 

3690 HALF7. = 2 

3700 GOTO 3360 

3710 CLOSE #1 

3720 REM END OF IOCONFIG.DAT ADJUSTMENTS 

3725 GOSUB 10000 : REM routine to set control parameters 

3727 GOSUB 12200 



PAGE 6 

3729 REM 

3730 REM ***** END OF PARAMETER SETUP ***** 
40fZ)(3 REM 

4001 REM ***** D.A.S„ SETUP ***** ^ 

4(302 CLS 

4003 LOCATE 13 

4004 PRINT "STANDBY WHILE KEITHLEY lOCHANNELS ARE SET UP." 

4009 CALL I NIT 

4010 VALUE =0' 

4011 VSTAT = 0' 

4012 PDT0FACT = 1 

4013 PT0FACT = 1 

4014 AT0FACT = 1/40.96 

4015 TT0FACT = 1 

4016 ATI FACT = .5/40.96 

4017 AT2FACT = 1/40.96 
4025 MEMSIZE = O: 
4030 HR"/. - 

4040 MIN7. = 

4050 SECy. =: 

4060 DATE/I = 

4070 MO"/. = 

4080 YR7. = 

40S5 STATUS7. = 

4090 CURDEP ' = 1 ' 

4092 QPENVALVE = 1 ' 

4094 CLOSEVALVE = 0' 

4100 OPEN "R" ,#1 , "IOCONFIG.DAT" ,72 

4110 FIELD #1 ,4 AS lOCARDt, 4 AS lOSLOT-f, 4 AS IOCHAN0:f, 4 AS IDCHANl;!:-, 4 ^^L , 

CHAN2*, 4 AS I0CHAN3:r-, 4 AS I0CHAN4:J?, 4 AS lOCHANSf-, 4 AS I0CHArT6:|-, 4 AS IC^Pf 

*,32 AS LASTS* 

4120 FIELD #1 ,40 AS FIRST 10*, 4 AS lOCHANS*, 4 AS I0CHAN9*, 1 AS I0CHAN13-F, 4 

3 lOCHANll*, 4 AS I0CHAN12*, 4 AS I0CHAN13*, 4 AS I0CHAN14*, 4 AS lOCHANlS* 

4130 FIELD #1, S AS FIRST2*, 64 AS LAST16* 

4140 FOR SLOT-/. = 1 TO 10 

4150 GET 1 ,SLOTy. 

4160 IF lOCARD* = "NONE" GOTO 4230 

4170 NUMCHAN7. = -1 

4180 FOR J = 1 TO 16 

4190 TEMP* = "lOCHAN" + STR*(J) + "*" 

4200 REM NOW REMO'-'E BLANK PUT IN BY STR* FUNCTION 

4210 CHANNAME*(J) = LEFT* ( TEMP* , 6) + MI D* ''TEMP* , S) 

4220 NEXT J 

4230 FOR J = 1 TO 16 

4240 IF MID*(LAST16*, (4*(J-1)+1) ,4) = "NONE" THEN J = 17 

4250 NUMCHAN7. = NUMCHAN7. + 1 

4260 NEXT J 

4270 IF LEFT*(IDCARD*, 1) = "A" THEN GOSUB 3700 ELSE 60SUB SB00 

4230 NEXT SL0T7. 

42<^0 CLOSE #1 

4300 REM MAKE BIT ARRAYS 

4310 CALL ARMAKE' ("0PEN5?<" , 1.0,5) 

4320 CALL ARMAKE ' ( " CL0SE3?- ",1.0,3) 

4330 CALL ARMAKE' ("0PEN2^<", 1.0,2) 

4340 REM FILL BIT ARRAYS ^ 

4350 FOR N7. = 1 TO 5 

4360 CALL ARPUTVAL ' ( "OPEN52<" , 1 . , N7. , 1 . 0) 



PAGE 7 

4370 NEXT N7. 

43S0 FOR N/l = 1 TO 3 

43<?0 CALL ARPlJT VAL ' • " CL0SE3.!< ",1 . Q , N7. ,0.0- 

m 4400 MEXT M'l 

44 1 CALL ARPLITVAL ' ( " OPEMZS-: ",1.0,1,1.0) 

4420 CALL ARPUTVAL ' ( "OPEN22<" , 1 . , 2 , 1 . 0) 

4430 REM INITIALIZE NUMTC7. (10) 

4440 FOR N = 1 TO 10 

4450 IF ( (LEN(TC^(N) )+l) MOD 5) - THEN GOTO 4510 ELSF I^ 'I EN ' TC >■- .'N ) ) Mf 
THEN GOTO 4530 ~ -um , i l,-. ij; ; mi 

4460 PRINT "THE TC LIST FOR REACTOR: " N " HAS THE ^'JPONG FORMAT " 

4470 PRINT "FORMAT IS: 4 LETTER TC NAME FOLLOWED BY ONE SPACE " 

44S0 PRINT "BEFORE NEXT 4 LETTER TC NAME. " 

4490 INPUT "HIT ENTER TO RETURN TO TC SETUP." PESPJf 

4500 GOTO 2430 ' 

4510 NUMTC7. (N) = (LEN f TC^ ( N) ) +1 ) / 5 

4520 GOTO 4540 

4530 NUMTC7. (N> = LEN(TC*<N)) / 5 

4540 NEXT N 

4550 REM CALCULATE TOTAL NUMBER OF -SAMPLES TO TAKE 

^570 REM ""'' ""'"■^ --''^ '"''''"' ^''^''^ '^ ^''°*'^^ space" IN MEMORY FOR MAIN ARRAV 

4580 REM 

4590 REM SET UP MAIN ARRAY 

4600 TOTTCy. = 

4610 FOR N = FIRSTREACT7. TO LASTREACTM 

4620 T0TTC7. = NUMTC7. (N) + T0TTC7. 

4630 NEXT N 

4o40 T0TREACT7. - LASTREACT7. - FIRSTREArT"/ + 1 

a!"n n^nv^^'^^o^^^"'- '^ " ^'^^'^ NUMTIMESTMP7. = 2 ELSE NUMTIMESTMRV. = 1 

4fc.^0 WID/. = T0TTC7. + (11 * T0TREACT7.) + (5 * NUMTIMESTMP7.) 

46o5 CLS: LOCATE 5 

4670 INPUT "ENTER TOTAL TIME FOR THIS RUN IN HOURS: " TOTTIMF^ 

4630 MEASTIME = (H20WAIT. / 60) . (C02WAIT>: / 60) . ^0^^'} ,0. -^i^,,TE 

4690 OTHERTIME = ' MINUTES/REACTOR FOR PROGRAM EXECUTION 

4700 SAMPTIME7. - (MEASTIME . OTHERTIME) * TOTREACTv hiVmTE- 

4710 IF SAMPTIME/: .; 30 THEN SAMPTIME:: = 30 MI,..UTE. 

4720 NUMSAMPS = ((TOTTIMEX * 60) / SAMPTIMF:-. + 1 

4730 MEMSIZE = FRE(0) 

4750 IF^SFFn^T;p''''''^S^''- * "'^"'- * ^ '^^° BYTES/WORD IN ARRAY 

4760 ^[g''^^^^^^^ ' -^^ * NEMSIZE THEN SAMPTIMEX ^ SAMPTIME7. . 10: GOTO 4^ 

4770 LOCATE 10 

R'-'TO^TIMEr-'SSCps: "''''""" ''''-'- '' '''^"^ '■' •' '^^'^'^^^^ " -^^^-^-^ INTERV 

mi ITJ-^L"''^^ ^'^"""^''^ ''''^'' ^■^^^^' TO CONTINUE. ",DUM 
4370 DIM MAIN7.(NUMSAMPS,WIDy.) 

4930 TMR = 60 ♦ HZ -veie 

5000 REM ♦*♦*♦ OPERATION ***** 
5010 CLS 



PAGE 3 

5020 LOCATE 3 

5030 INPUT "TYPE 'YES' TO BEGIN RUN,, MO' TO ABORT NOKl: " , RESP'J? 

5040 IF RESP* = "YES" GOTO 5042 ELSE IF RE3P* -■= "NO" QQTO 7'v'?'? ELSE GOTO 5f|i.0 

5 4 2 R E N r-'l a i n d a t a a c q u. i s i t. i o n 1 d d p A 

5043 60SUB V200 PRINT HEADER ON THE PRINTER 

5050 CLS 

5052 PRINT "# SAMPLES:" TAB (50) "ELAPSED TINE": PR I NT SPC(30) "REACTOR #" 

5054 PRINT TAB(10) "1" TAB(16) "2" TAB(22) "3" rAB(2S) "4" Th6(34) "5" TAB(4Q) 
6" TAB(4A) "7" TAB (52) "8" TAB(5S) "9" TAB (63:' "10" 

5056 REN DATA TEl . PT0 , TT0 , AT0 , AT2 , AT 1 , POT0 , PT0 , TTQ , AT0 , AT2 , AT I 

5055 PRINT "INLET VALUES" 

5060 FOR J = 1 TO 12: READ NEAS* ( J ) : NEXT J 

5062 FOR J - 1 TO 6 

5064 PRINT USING "\ \"; MEAS$(J): NEXT J 

5065 PRINT "OUTLET VALUES" 

5066 FOR J = 7 TO 12: PRINT USING "\ \"; MEAS:f(J) : NEXT J 

5067 INITMO:.?^ = LEFT*(DATE-t,2) 
506S INITDAY* = NID* (DATE* , 4 , 2) 

5069 INITTIME = VAL (MID* ( T IME* , 4 , 2) ) + (VAL (LEFT* (T IME* , 2) ) * 60) 

5070 FOR CURDEP = 1 TO NUMSAMPS 

5072 60SUB 9300 'CALCULATE ELAPSED TINE 
5075 COLR = COLR XOR 1: COLOR (3 + C0LR*2) 
5077 LOCATE 1,63: PRINT ELAPHOUR ":" ELAPNIN 

5073 LOCATE 1,12: PRINT CURDEP 
5079 PLACE% = 1 

50S0 GOSUB 3500 

5090 FOR I = FIRSTREACTy„ TO LASTREACTV. 

5100 IF I = FIRSTREACT7. + 4 THEN GOSUB 3500 

5110 FOR l< === 1 TO STEP -1 ^ 

5115 IF K = THEN PRINT M 

5120 VALVE* - SV*(K,I) 

5130 VSTAT --= OPENVALVE 

5140 GOSUB 8000 

5150 IF K = GOTO 5270 

5160 REM Read TC ' s ior this reactar 

5165 REF* = "REF3 " 

5166 IF I > 3 THEN REF* = "REF4 " 
5170 PARAMLI3T* = REF* + TC* ( I ) 
5130 GOSUB 3300 

5190 REN Take each value from "paramsVu" and put it in "main"'," 

5200 FOR J = 1 TO NUMTCV. ( I ) 

5201 PRINT #3, "TEMPS "; 
5210 WID7„ = J + 1 

5220 CALL ARGETVAL ' ( " PARAMSV. " , 1 . , W 1 07. , VALUE ,13) 

5224 IF J <> 1 GOTO 5230 

5225 LOCATE 5,((I*6)+1): PRINT USING "##.#"; VALUE 
5230 MAIN'/. (CURDEP, PL ACE"/.) = VALUE * 10 

5236 PRINT #3, USING "####. tt" ; VALUE; 

5240 PLACEV. = PLACE7. + 1 

525.0 NEXT J 

5251 PRINT #3, 

5260 CALL ARDEL ' ( "PARAMS7." ) 

5270 IF K = 1 THEN PRINT #3,1 " INLET " ; ELSE PRINT #3,1 " OUTLET "; 

5271 IF K = 1 GOTO 5279 

5272 PARAM* = "PDTO" M 



PAGE 9 

5273 REM Read PDTO 

5274 PARAMO = "PDT0" 

5275 60SUB 8200 'read one analog point 
527A VALUE = (VALUE-2048) ^ PDT0FACT 

5277 GOSUB S400 write a value to main array 
5279 REM Read PTO 
5230 PARAM* = "PTO" 

5290 GOSUB 3200 'read one analog point 

5291 VALUE = fVALUE-2048) * PTOFACT 

5300 GOSUB 3400 'write a value to main array 
5310 VALVE* = "SV8" 
5320 VSTAT = OPENVALVE 
5330 GOSUB 8000 'open valve SV8 
5340 CALL PAUSE' (H2nLgA IT-/., "SEC") 
5350 PARAMO = "TTO" 

5360 GOSUB 8200 'read H2n sample temp 
5365 VALUE = (VALUE-2048) * TTOFACT 
5370 GOSUB 8400 
5380 PARAMJ: = "AT0" 
5390 GOSUB 8200 'read humidity 
5395 VALUE = (VALUE-2048) * ATQFACT 
5400 GOSUB 8400 
5410 VSTAT = CLCSEVALVE 
5420 GOSUB 8000 'close valve SV3 
5430 VALVEf - "SV10" 
5440 VSTAT = OPENVALVE 
5450 GOSUB 8000 'open SVIO 
5460 CALL PAUSE ' (Cn2WHl T7. , "SEC" ) 
5470 PARAM* = "AT2" 
15430 GOSUB 3200 'read 7„C02 
5485 VALUE = (VALUE-204a) * AT2FACT 
54^0 GOSUB 8 400 
5500 VSTAT = CLOSEVALVE 
5510 GOSUB 8000 'close SVIO 
5520 REM section to read "/.02 in gas stream 
5530 VSTAT = CLOSEVALVE 
5540 VALVE-r = "SVll" 

5550 GOSUB 8000 CLOSE SVll 

5560 VALVES-f = "OPENS?.:" 
5570 VALVELIST* = 02STRTLI3T-f 
5580 GOSUB 8100 
55''0 PARAMl: = "PTl" 
5600 TEMPO = -1 ' 
5610 TEMPI = -2' 
5620 WHILE TEMPO :> TEMPI 
5630 GOSUB 8200 
5640 TEMPO = TEMPI 
5650 TEMPI = VALUE 
5660 CALL PAUSE '< 1 , "SEC" ) 
5670 WEND 
5680 PTl VALUE = VALUE - 1' 

5690 VALVES* = "CL0SE3.!<" 

5691 VALVELIST* = 02STRTPT2-f 

5692 GOSUB 8100 

.5700 REM Routine to match PT2 to PTl 



PAGE li/D 

5710 CALL SCHMITRIG' ( "PT2" , -1 . Q „ PT 1 VALUE „ "ABOVE "„ "ST" - 

5720 CALL D I GWR I TE ' ( " SV5 " , . (3 , " WST " ) 

5730 REM Get ready to read 02 

5743 VALVES* = "0PEN2^;" 

5741 VALVELIST* = n2REHDY^? 

5742 GOSUB 8100 

5750 CALL PAUSE ' ( 02WA I T/l , " SEC " ) 

5760 PARAM* = "ATI" 

5770 GOSUB 8200 'read /:D2 

5775 VALUE = (VALUE-2043) * ATI FACT 

5730 GOSUB 8400 

5790 VALVES* = "CLQSE3?<" 

5791 VALVELIST* = 02END* 

5792 GOSUB 8100 

5800 VSTAT = OPEMVALVE 

5810 VALVE* = "SVll" 

5820 GOSUB 3000 'OPEN SVll TO EMPTY STORAGE BOTTLES 

5830 REM End of 7.02 section 

5340 VALVE* = SV*(K,I) 

5350 VSTAT - CLOSEVALVE 

5860 GOSUB S000 

5362 PRINT 1+3, 

5864 NEXT K 

5865 REM routine to initiate control 
5369 GOSUB 1100Q 

5871 GOTO 5880 'TEMP FOR TESTING 

5372 CURMIN = VAL (MID* ( T IME* , 4 , 2) ) + < VAL (LEFT* ( TIME* , 2) ) * 60.: 

5373 IF CURMIN < STRTMIN GOTO 5880 

5874 IF STRTMIN + SAMPTIMEX > CURMIN GOTO 5872 
5880 NEXT I 

6998 PRINT #3, 

6999 NEXT CURDEP ! 

7000 REM END OF MAIN DATA ACQUISITION LOOP 
79^9 PRINT "AT LINE 7999 ": END 

8000 REM open /close valve 

3010 CALL DIGWRITE' (VALVE*, VSTAT) 

S020 RETURN 

3100 REM open/close a set of valves 

3110 CALL D I GOUT ' ( VALVES* , VAL VEL I ST* ., 1 , " DGTLOUT " ) 

8120 CALL INTON ' ( 10, "MIL" ) 

3130 CALL STATUS' ("DGTLOUT" ,STATUS7.) 

3140 IF STATUS^ = 1 GOTO 3130 

8150 CALL INTOFF 

3190 RETURN 

3200 REM Read one analog input 

32 1 CALL ANREAD ' ( P ARAM* , VALUE ) 

3220 RETURN 

8300 REM Read a set of analog inputs 

33 1 CALL AN I N ' ( " PARAMS"/. ",1.0, PARAML I ST* , 1 , " ANALOG IN") 

3320 CALL INTON' (30, "MIL") 

3330 CALL STATUS '( "ANALOGIN" , STATUS7.) 

3340 IF STATUS'/. = 1 GOTO 3330 

3350 CALL INTOFF 

8390 RETURN 

3400 REM Write a value to main array ?< to printer ?< to screen 

3410 MAIN'/. (CURDEP, PLACE'/.) = VALUE 

8412 IF NEWLINE = 1 THEN PRINT #3,: NEWLINE = 



PAGE 11 

3413 PRINT #3, USING "####. tt" ; VALUE; 

3414 IF NOSCREEN = 1 GOTO 3480 

3415 LOCATE ,((I*6)+1): PRINT USING "##. tt" ;; VALUE 
3 430 PL''-CE".'. = PLACE". + '. 

3481 NOSCREEN = 

8490 RETURN 

3500 REN Place time stamp in main array 

3510 CALL CLOCKREAD' (HPr. , NIN7. , SEC7. , DATE"/. , NOV. , YR'/.) 

3520 VALUE = HR7. 

3525 NOSCREEN = 1 

8530 60SUB 8400 

3540 VALUE = MIN7. 

3545 NOSCREEN = 1 

3550 GOSUB 3400 

8560 VALUE = NO/l 

3565 NOSCREEN = 1 

3570 GOSUB 3400 

3580 VALUE = DATE7. 

3585 NOSCREEN = 1 

3590 GOSUB 3400 

3600 VALUE = YR7. 

3605 NOSCREEN = 1 

3610 GOSUB 3400 

8619 PRINT #3," SAMPLE # " CURDEP 

8620 RETURN 

3700 REM SET UP lONANES FOR ONE ANALOG CARD 
3^02 IF lOCHANO:!^ = "NONE" THEN GOTO 8704 
8703 CALL lONAME ' ( lOCHANO* , SL0T7. , , 1 2) 
3704 IF lOCHANll- = "NONE" THEN GOTO 3706 
I 3705 CALL lONANE ' (I0CHAN1^-,SL0T7. , 1 , 12) 

3706 IF I0CHAN2* = "NONE" THEN GOTO 3703 

3707 CALL lONAME ' (I0CHAN24: , SL0T7. , 2 , 12) 
8708 IF I0CHAN3J: = "NONE" THEN GOTO 3710 
870'' CALL lONANE ■ ( I0CHAN3f-,SL0T"/.,3, 12) 

8710 IF I0CHAN4f = "NONE" THEN GOTO 3712 

8711 CALL lONANE' (I0CHAN4^,SL0T7.,4, 12) 

8712 IF I0CHAN5-f- = "NONE" THEN GOTO 37 14 

8713 CALL lONAME' (I0CHAN5:$:,SL0T7.,5, 12) 

8714 IF I0CHAN6* = "NONE" THEN GOTO 3716 

8715 CALL IDNANE' (I0CHAN6J:,SL0T-/:,6, 12) 

8716 IF I0CHAN7^ = "NONE" THEN GOTO 3713 

8717 CALL lONANE ' ( I0CHAN7:r ,SL0T7. ,7, 12) 
3718 IF lOCHANG^ = "NONE" THEN GOTO 3720 
8719 CALL lONANE ' n0CHAN3-t:,SL0T7. ,8, 12) 
3720 IF I0CHAN9# = "NONE" THEN GOTO 8722 
8721 CALL lONAME' (I0CHAN9:$,SL0T7.,9, 12) 
3722 IF IOCHAN10* = "NONE" THEN GOTO 8724 

8723 CALL I DNANE ' ( I OCHAN 1 0:*: , SL0T7. ,10,12) 

8724 IF lOCHANllf = "NONE" THEN GOTO 3726 

8725 CALL lONAME ' ( lOCHANl 1 :? , SL0T7. , 1 1 , 12) 

8726 IF I OCHAN 12* = "NONE" THEN GOTO 8723 

3727 CALL lONAME ' ( lOCHANl 2* , SL0T7. , 1 2 , 12 ) 

3728 IF iaCHAN13* = "NONE" THEN GOTO 3730 

3729 CALL lONAME ' (I0CHAN13J: , SLOTV. , 13 , 1 2) 
^8730 IF I0CHAN14* = "NONE" THEN GOTO 8732 
r3731 CALL lONAME' (I0CHAN14f ,SL0T7., 14, 12) 



F'AGE 
3732 
3733 
3734 
3735 
3736 
3760 
3300 
3302 
3303 
3304 
3305 
3306 
3807 
3303 
3309 
3310 
3311 
3312 
3313 
33 1 4 
3315 
3316 
3317 
3313 
331? 
3320 
3321 
3322 
3323 
3324 
3325 
3326 
3327 
3323 
3329 
3330 
3331 
3332 
3333 
3360 
9000 
9010 
9020 
9030 
9040 
9050 
9071 
9060 
9061 
9062 
9063 
9064 
9065 
9066 
9067 
9063 



12 

IF I0nHAM15* = "NONE" THEN GOTO 3734 
CALL I OMAME ' <■ I OCHAN 1 5* , SLOT"/. ,15,12) 
IF IOCARD$ <> "AIM7" THEN GOTO S760 
REF:S = "REF" + CHRi-lJ(SL0TX + 43) 
CALL I ONAME ' ( REF* , SLOTX , 32 , 1 2 ) 
RETURN 

REM SET UP lONAMES FOR ONE DIGITAL CARD 
IF inCHAN0* - "NONE" THEN GOTO 8304 
I OCHAN0* , SLOT/: , ) 
"NONE" THEN GOTO 3306 



CALL lONAME 
IF lOCHANlJF 
CALL I ONAME ' ( I OCHAN 1 * , SLOT": , 1 



IF I0CHAN2-* 



"NONE" THEN GOTO 3303 



CALL I ONAME ' ( I 0CHAN2f: , SLOT"/. , 2 ) 



IF I0CHAN3* 

CALL I ONAME 

IF I0CHAN4:|: 

CALL I ONAME 

IF I0CHAN5* 

CALL I ONAME' (lOCHAh 

IF I0CHAN6* 

CALL I ONAME 

IF I0CHAN7$ 

CALL I ONAME 



= "NONE" THEN GOTO 3310 

(I0CHAN3*,SL0T-/. ,3) 

= "NONE" THEN GOTO 3312 

(1 0CHAN4* , SL0T7. , 4 ) 

= "NONE" THEN GOTO 3314 
;ir,SLOT"/. ,5) 

= "NONE" THEN GOTO 3316 

( 1 0CHAN6* , SLDT7. , 6 ) 

= "NONE" THEN GOTO 3313 

a0CHAN7*,SL0T-/.,7) 
IF IQCHAN3* = "NONE" THEN GOTO 3320 
CALL I ONAME ' ( I OCHANS* , SLOT X , 3 ) 
IF I0CHAN9:if = "NONE" THEN GOTO 3322 
CALL I ONAME ' < 1 0CHAN9:|: , SLOT/l , 9 ) 
IF I OCHAN 10* = "NONE" THEN GOTO 3324 
CALL I ONAME ' ( I OCHAN 1 0$ , SLOT /l ,10) 
IF I OCHAN 11* = "NONE" THEN GOTO 3826 
CALL I ONAME ' ( I OCHAN 11*,, SLDTy. ,11) 
IF I OCHAN 12* = "NONE" THEN GOTO 3323 
CALL I ONAME ' ( I OCHAN 1 2* , SLOTV- ,12) 
IF I OCHAN 13* = "NONE" THEN GOTO 3330 
CALL I ONAME ' ( I OCHAN 1 3* , SLOT a , 1 3 ) 
IF I OCHAN 14* ^ "NONE" THEN GOTO 3332 
CALL I ONAME ' ( I OCHAN 1 4* , SL0T7. ,14) 
IF I OCHAN 15* = "NONE" THEN GOTO 3360 
CALL I ONAME ' ( I OCHAN 1 5* , SLOTZ ,15) 
RETURN 

REM ROUTINE TO UPDATE A SPECIFIC INSTRUMENT 
REM NAME IN FILE IOCONFIG.DAT 

REM SLOTy. HAS # OF SLOT, CHANV: HAS # OF CHANNEL 
REM AND I OCHAN* HAS NAME OF INSTRUMENT 
GET 1„SL0T7. 

ON (CHAN/L + 1) GOTO 9060, 906 1 , 9062 , 9063 , 9064 , 906^ 
9072 , ^073 , 9074 , 9075 



?066 , 9067 , <?063 „ 9069 , 90: 



LSET IOCHAN0* 

LSET I OCHAN 1* 

LSET I0CHAN2* 

LSET I0CHAN3* 

LSET I0CHAN4* 

LSET I0CHAN5* 

LSET I0CHAN6* 

LSET I0CHAN7* 

LSET I0CHAN3* 



I OCHAN* :GOTO 9030 

I OCHAN* :GOTO 9030 

I OCHAN* :GOTO 9030 

I OCHAN* :GOTO 9030 

I OCHAN* :GOTO 9080 

I OCHAN* :GOTO 9030 

I OCHAN* :GOTO 9030 

I OCHAN* :GOTO 9030 

lOCHAN* :60T0 9030 



lOCHANit^ :GOTO 9080 

: lOCHANJ: :GOTO 9030 

■ lOCHAN* :60T0 90S0 

■ lOCHAMii^ :GOTn q>0ao 
: lOCHAN*: .-GOTO 91.330 
: IOCHAN:|: :60T0 9030 

lOCHAM:!: :GOTO 90S0 



PAGE 13 

9069 LSET IOCHAN=i>J: = 

9070 LSET IQCHANlOl: 
''OTl LSET lOCHANll* 
■^072 LSET inCHAN12l: 
9073 LSET I0CHANi3-r 
^'074 LSET I0CHAM14:f 
9075 LSET I0CHAN15* 
9080 PUT 1 , SL0T7. 
9090 RETURN 

9200 REN ROUTINE TO PUT INITIAL HEADER ON PRINTER 

'7210 OPEN "LPTl:" FOR OUTPUT AS #3 

9220 PRINT #3, CHR-^(12) "Run beginning on " DATE-f " 

9230 PRINT "Enter identification info, for this run 

9240 INPUT DUN* 

9250 IF LEN'iDUM-J:) = GOTO 9270 

9260 PRINT #3, DUN#: GOTO 9240 

9270 PRINT #3,: PRINT #3, TAB (16) "PDTO PT0 



TT0 



" TINE* 
empty line 



AT0 



ATI 



60) 



<52S0 RETURN 

•^300 REN ROUTINE TO CALCULATE ELAPSED TINE 

9310 IF LEFT* (DATE*, 2) = INITNO* GOTO 9340 

•^320 INITTINE = INITTINE - 24*60 

9330 INITNO* = LEFT*(DATE*,2) : INITDAY* = "1" 

9340 IF NID*(DATE*,4,2) = INITDAY* GOTO 9400 

9350 INITTINE = INITTINE - 24*60 

9360 INITDAY* = NID* (DATE* , 4 , 2) 

<?400 STRTNIN = VAL > N ID* ( T INE* , 4 , 2 ) ) 

<?410 EL APT I NE = STRTNIN - INITTINE 

"^420 ELAPHOUR = ELAPTINE\60 

'7430 ELAPNIN = ELAPTINE NOD 60 

■7440 RETURN 

10000 REN 

10010 REN 

10020 REN 

10022 DIN CV* ( 10,5) ,CV1 ( 10) 

10024 OPEN "I" , 1 , "CNTVALVE.DAT" 
10026 FOR X = 1 TO 10 

10025 INPUT #1 ,CV*(X, 1) ,CV1 (X) ,CV*(X,2) ,CV*(X,3) ,CV*(X,4) ,CV*(X,5) 
10030 NEXT X 

10032 CLOSE 1 

10040 REN 

10050 CLS:I<EY OFF 



(VAL (LEFT* (TINE*, 2) ) '■ 
'VALUE IN NINUTES 
'TRUNCATE TO INTEGER 
'RENAINDER IS NINUTES 



10060 LOCATE 1 ,32 
10070 LOCATE 2,23 
10072 LOCATE 3,23 



PRINT "valve control menu' 
PRINT"A BCD 
PR I NT "1 2 3 4 



10080 LOCATE 4,1: PRINT "measurement device" 

10085 LOCATE 5,1: PR INT " measured on:" 

10090 LOCATE 6,1:PRINT "trigger value" 

10100 LOCATE 3,1:PRINT "trigger select" 

10110 LOCATE 10,1:PRINT "trigger on/off" 

10115 LOCATE 12,1:PRINT "valve to activate" 

10160 Z = 16 

10165 LOCATE 4,20 

10170 FOR X = 1 TO 10 



PAGE 1 

10130 

10190 

10192 

ISl-^A 

10196 

1019S 

1 0200 

10232 

1 020S 

10210 

10215 

10220 

10230 

10240 

1 0250 

10260 

10270 

10275 

10230 

10290 

10300 

10310 

10320 
10330 
10332 
10334 
10336 
10333 
1 0339 
10340 
10345 
10350 
10360 
10370 
10440 
10450 
10455 
10460 
10462 
10464 
10463 
1 0470 
10480 
104'^0 
10500 
1 05 1 
10520 
10530 
10540 
10550 
10560 
10570 
10530 
105S2 
10590 
10600 



PRINT ThB(Z) ;Cy:J:(X, 1) ; 
NEXT X 

LOCATE 5,20 
FDR X = 1 TO 10 

2 = Z + 6 

PRINT TAB(Z) ;CV*(X,5) ; 
NEXT X 
Z = 16 
LOCATE 6,20 
FOR X = 1 TO 10 

Z = Z + 6 

PRINT TAB(Z) ; 

PR I NT US I NG " ## . ## " ; CV 1 ( X ) 5 
NEXT X 
Z = IS 
LOCATE 3,20 
FOR X = 1 TO 10 

Z = Z + 6 

PR I NT TAB ( Z ) ; CV^t ( X , 2 ) 5 
NEXT X 
Z = 18 

LOCATE 10,20 
FOR X = 1 TO 10 

Z = Z + 6 

PR I NT T AB ( Z ) ; C'J^ ( X , 3 ) 5 
NEXT X 

LOCATE 12,20 
Z = 16 
FOR X = 1 TO 10 

Z = Z + 6 

PR I NT TAB ( Z ) ; CV* ( X , 4 ) ; 
NEXT X 

LOCATE 14,10:PRINT "THESE ARE YOUR CURRENT SETTINGS" 
LOCATE 15, 10: INPUT "DO YOU WISH TO MAKE ANY CHANGES CN: 



IF Zl* 
IF 11$ 
IF Zl* 
PRINT 
LOCATE 
LOCATE 13 
IF Z2 
LOCATE 



= "" THEN Zl* = "N" 

= "N" OR Zl* = "n" THEN 10963 

="Y" OR Zl* = "y" THEN 10463 

RESPONSE I NCORRECT , TRY AGA I N " : GOTO 1 0440 

13,1: FOR X = 1 TO 10: PRINT TAB (79);" ":NEXT ; 

SELECT FERMENTER # " ; Z2 

THEN 10510 

THAT IS AN INCORRECT VALUE , TRY 



10 : INPUT 
= 10 AND Z2 
15,10 : PR I NT 
GOTO 10470 
LOCATE 13,1: FOR X = 
LOCATE 13,27: PRINT 
LOCATE 14,20: PRINT 
LOCATE 15,20: PRINT 



PRINT USING "##.##";CV1( 



LOCATE 16,20: PRINT 
LOCATE 17,20: PRINT 
LOCATE 13,20: PRINT 
82 LOCATE 19,20: PRINT 
LOCATE 20,1 : INPUT 
IF Z3 >=1 AND Z3 <> 



1 TO 11:PRINT TAB (79) : 
FERNENTER # "; Z2 

1. MEASUREMENT DEVICE 

2. TRIGGER VALUE 



3. TRIGGER SELECT 

4. TRIGGER ON /OFF 

5. VALVE TO ACTIVATE 

6. EXIT" 
SELECT VALUE" ;Z3 

6 THEN 10630 



IE XT 



CV*(Z2, 1) 



; CV* (Z2,2) 
;CV*(Z2,3) 
;CV*(Z2,4) 



:CV*(Z2 



PAGE 1 
10610 
10620 
1 0630 
10632 
10634 
10636 
10637 
1063S 
10639 
10640 
10645 
10647 
10650 
10652 
10660 
10670 
10680 
106^0 
10700 
10710 
10720 
10730 
10740 
10750 
10760 
10770 
10730 
10790 
10S00 
10310 
10315 
10320 
10830 
10840 
10850 
10860 
10368 
10870 
10830 
10882 
10884 
10890 
10900 
10910 
10912 
10914 
10916 
10918 
10920 
10922 
10924 
10926 
10928 
10930 
10940 
> 10964 



PRINT:PRir4T "VALUE INCORRECT TRY AGAIN" 

GOTO 10590 

IF Z3 <> 1 THEN 10700 

LOCATE 21,1: PR I NT "MEASUREMENT OPTIONS: " 

LOCATE 22, 5: PR I NT "THERMOCOUPLES: "; 

COLOR 14: PRINT TC*(Z2) 

COLOR 2:L0CATE 23,5:PRINT "OTHER DEVICES AVAILABLE: ":: COLOR 14 

PRINT"PTDO PTO 02TP H207. C02X 027.": COLOR 2 
REM 

LOCATE 20, 1 

INPUT" MEASUREMENT TYPE ;^: INPUT (IN) OR OUTPUT (OUT) MEASUREMENT": 

IF MID*(Z4*, 1 , 1) = "T" OR MID* ( Z4:|^ , 1 , 1 ) = "t" THEN 25^ - "IN" 

IF LEN(Z4*) <> 3 AND LEN(Z4*:) <> 4 THEN 10660 
IF Z5* = "IN" OR Z5:r = "in" OR Z5* = "OUT" OR 25^ = "out" THEN 1363t; 

LOCATE 24, 1:PRINT"VALUE(S) INCORRECT , USE FOUR PLACE CODE. IN OR OUI 

GOTO 10632 

CV*(Z2,1) = Z4-* :CViMZ2,5) = ZS* 
REM 
IF Z3 <> 2 THEN 10760 

INPUT "ENTER NEW TRIGGER VALUE" ;Z5 

REM 

REM 

CVl (Z2) = Z5 

REM 
IF Z3 <> 3 THEN 10350 

LOCATE 20, 1:PRINT"CH00SE NEW TRIGGER SELECT" 

PRINT "CHOOSE >= WHEN YOU WANT TO ACTIVATE VALVE ABOVE OR EQUAL TC 

PRINT "TRIGGER VALUE OR < WHEN ACTIVATION IS LESS THAN TRIGGER VAL 



THEN 10340 



= 1 TO 

'ENTRY 



3: PRINT TAB (79) 5" ":NEXT X 
INCORRECT USE EITHER >= OR 



IF Z4.f 
IF Z4* = 
IF 14$ = 



INPUT ">= OR <":Z4:i: 
IF Z4:$: = '•>=" OR Z4:j: = "< 
LOCATE 20, 1:F0R X 
LOCATE 19,1: PRINT 
GOTO 10770 
CV::E(Z2,2) = Z4:f 
REM 
IF Z3 <> 4 THEN 10914 

LOCATE 20,1: PRINT TAB(79);" " 

LOCATE 20, 1: INPUT "SELECT TRIGGER ON , OFF , NA" ; Z4:F 
•ON" OR Z4* = "OFF" THEN 10910 
'on" OR Z4* = "off" THEN 10910 
■na" OR 14$ = "NA" THEN 10910 
PRINT "VALUE INCORRECT USE EITHER ON OR OFF" 
GOTO 10363 
CV:$:(Z2,3) = Z4# 
REM 

IF Z3 <> 5 THEN 10964 
LOCATE 20,1: PRINT TAB (79);" " 
LOCATE 20,1: INPUT "VALUE TO ACT I VATE" ; Z4* 
IF LEN(Z4t:) = 4 THEN 10940 

PR I NT: PR I NT "VALUE INCORRECT, IT MUST BE A 4 PLACE CODE 
GOTO 10916 
REM 
REM 
REM 

CV*(Z2,4) = Z4-1: 
IF Z3 <> 6 THEN 10510 : REM PRINT NEW FERMENTER MENU 



PAGE 16 

10966 BOTO 1(3050 : REM PRINT NEW SCREEN 

10963 OPEN "o" , 1 , "cntvalve.dat" 

10970 FDR X = 1 TO 10 

10972 PRINT #KCV*^X,1) 

10974 PRINT #1,CV1 (X) 
10976 PRINT #1 ,CV-l;(X,2} 

1 0975 PR I NT # 1 , CV* ( X , 3 ) 
1 0979 PR I NT # 1 , C^J.^ ( X , 4 ) 
1 0930 PR I NT # 1 , C V* ( X , 5 ) 
10933 NEXT X 

10990 CLOSE 1 

10992 RETURN 

11000 REM BEGIN VALVE CONTROL ROUTINE 

11002 REM CHECK TO DETERMINE IF MEASUREMENT I'^ INLET OR OUTLE" 

1 1 004 FUD = 

11006 IF CV$(I,5) = "IN" OR CV*(I,5) - "m" THEN FUD - 6 

11010 YY 1 = LEN ( TC* ( I ) ) 

11020 REM COUNT NUMBER OF THERMOCOUPLES STnpED TN ARRAY 

11030 CT - 1 

11040 FOR Y21 = 1 TO YYl 

1 1 0r50 AA:f = M I D* ( TC^t: (I ) , Y Z 1 , 1 ) 

11060 IF AA:iF = " " AND YZl > 4 THEN CT - CT + 1 

11070 NEXT YZl 

11030 IF YYl < 4 THEN CT = 

11090 REM CHECK THERMOCOUPLE ARRAY FOR MEASUREMENT TYPE MATCH 

11394 CT - CT -1 

11100 ST = 1 

11110 TCP = 

11115 TCPOS = 

11120 FOR ZZ = 1 TO CT 

11125 TCP - TCP + 1 

11130 B* = MID;* (TCt ( I) ,ST,4) 

11140 IF B$ = CV:f (I , 1) THEN TCPOS = TCP 

11155 ST = ST + 5 

11160 NEXT ZZ 

11130 REM SELECT PROPER READING FROM MA I NX ARRAY 

11190 IF TCPOS = THEN 11250 

11200 REM THERMOCOUPLE READINGS SELECTED 

11210 REM CALCULATE DATA POSITION SPECIFIC TO TERMOCOUPLE 

11220 RELPOS = TCPOS - CT -7 -FUD 

11230 ABPOS = PLACE"/. + RELPOS 

11240 GOSUB 12000 

11245 GOTO 11490 

11250 REM SELECT WHEN PTDO 

1 1 260 I F CV* (1,1) < > " PTDO " THEN 1 1 300 

11270 ABPOS = PLACE"/. - 6 -FUD 

11230 GOSUB 12000 

11290 GOTO 11490 

11300 REM SELECT WHEN PTO 

11310 IF CV$(I,1) <> "PTO" THEM 11340 

11320 ABPOS = PLACE"/. - 5 -FUD 

11330 GOSUB 12000 

11335 GOTO 11490 

11340 REM SELECT WHEN OXYGEN TEMP (02TP) 

11350 IF CV.t(I,l) <> "02TP" THEN 11330 

11360 ABPOS = PLACE-/.-4 - FUD 

11370 GOSUB 12000 



PAGE 17 

11375 GOTO 11490 

11380 REM SELECT WHEN PERCENTAGE OF HUMITITY (H207.) 

11370 IF CV-^(I,1) <> "H207." THEN 11420 

; 1400 ABPns = PLACE'.: - 3 - FIJD 

11410 G03UB 12000 

11415 GOTO 11490 

11420 REn SELECT WHEN PERCENTAGE OF C02 (CQ27,) 

1 1430 IF CV* (1,1) < > "C027." THEN 1 1 460 

11440 ABPOS = PLACE7. - 2 - FUD 

11450 GOSUB 12000 

11455 GOTO 11490 

11460 REM SELECT WHEN PERCENTAGE 02 

11465 IF CV*(I,1) •:;:> "027." THEN 11485 

11470 ABPOS = PLACE7. -1 - FUD 

11480 GOSUB 12000 

11482 GOTO 11490 

11485 PRINT "ERROR" 

11490 RETURN 

12000 REM ROUTINE TO CHECK IF VALVE SHOULD BE TRIGGERED 

12010 AAl = MA I N7. (CURDEP, ABPOS) 

12015 PRINT "VALUE "; AAl ; "CURDEP" ; CURDEPj "ABPOS" ; ABPOS 

12020 IF CV:|:':i,2) = ">=" THEN 12050 

12030 IF AAl < CVl(I) THEN GOSUB 12100 

12040 GOTO 12060 

12050 IF AAl >= CVl(I) THEN GOSUB 12100 

12060 RETURN 

12100 REM ROUTINE TO TRIGGER VALVE OPEN OR CLOSE 

12110 VALVES = CV-^ (1,4) 

12120 IF CV|:(I,3) = "ON" OR CV*-'I,3) - "on" THEN VSTAT - OPENVALVE 

12130 IF CV*(I,3) = "OFF" OR CV:.1?(I,3) = "off" THEN VSTAT = PLOSEVALVE 

12140 IF CV*(I,3) - "NA" OR CV:f(I,3) - " na" THEN GOTO 12163 

12143 GOSUB 9300 

12145 IF TMR > ELAPTIME THEN 12160 

12150 GOSUB 8000 

12160 RETURN 

12200 REM - routine to setup timing sequence 

12202 DIM ZD-^ ( 10,7) : KEY OFF 

12204 OPEN "i",l , "timer.dat" 

12206 FOR X = 1 TO 10 

12208 FOR Y = 1 TO 7 

12210 INPUT #1 ,2D:$:(X,Y) 

12212 NEXT Y 

12214 NEXT X 

12216 CLOSE 

12220 CLS:LOCATE 2, 1 

12225 TB = 3 

12230 PRINT TAB(3) ; "FERMENTERS" 

12240 LOCATE 4, 1 

12245 CR = 64 'set Initial ASCII value 

12Z50 FOR X = 1 TO 10 

12260 CR = CR + 1 

12270 IF X = 9 THEN CR = CR + 1 

12275 IF X = 10 THEN TB = 2 

12230 PRINT TAB(TB) ;X;CHRJ-(CR> 

12290 NEi<T X 



PAGE 


iS 


12300 


REM 


12310 


LOCATE 1,23 


12320 


PRINT "TIMING SEQUENCE VALVE TO CONTROL SEQU 


12330 


'"' - 4 ' 3et initial position for printinq 


12340 


FOR X - 1 TO 10 


12350 


LOCATE Y, 22: PRINT "- MIN - MIN" ; TAB i 65) ^ " 


12355 


Y = Y + 1 


12360 


NEXT X 


12435 


Y = 1 


12440 


FOR X = 4 TO 13 


12450 


LOCATE X , 1 2 : PR I NT Z D$ ( Y , 1) 


12460 


LOCATE X , 1 9 s PR I NT 2D* ( Y , 2 ) 


12470 


LOCATE X, 23: PR I NT ZD*(Y,3) 


1 24S0 


LOCATE X, 31 SPRINT ZD-4r(Y,4) 


12490 


LOCATE X, 35: PR I NT ZD*(Y,5) 


12500 


LOCATE X, 53 SPRINT ZD*(Y,6) 


12510 


LOCATE X, 65 SPRINT ZD*(Y,7) 


12515 


Y = Y + 1 


12520 


NEXT X 


12530 


LOCATE 15,5 


12540 


INPUT "These are your current sett i ngs , chanqe ^ Y or N 


12545 


IF 2Z* = "" THEN RETURN 


12550 


IF ZZ-* = "N" OR ZZ$ =■• "n" THEN RETURN 


12555 


LOCATE 1 5 , 1 : PR I NT TAB ( 7'=> ) ; " " 


12560 


LOCATE 15,5s INPUT" enter -fermenter number (l-10)"sZT 


12570 


IF ZT < 11 AND ZT > THEN 12590 


125S0 


LOCATE 16, 5: PRINT " invalid entry, try again "s GOTO 12 


12590 


LOCATE 1 5 , 1 : PR I NT TAB ( 73 ) ; " " 


12600 


LOCATE 16,1: PRINT TAB (73);" " 


12610 


LOCATE 16,1:PRINT TAB ( 34) 5 "FERMENTER #";ZT 


12620 


LOCATE 17, Is PRINT TAB(26)s"l- Activate": 


12630 


LOCATE 13,1 SPRINT TAB (26); "2. Timing sequence" 


12640 


LOCATE 19,1: PRINT TAB (31 ) ; "Fi r st " 


1 2650 


LOCATE 20,1 SPRINT TAB (31 ); "Second " 


12660 


LOCATE 21,1 SPRINT TAB(26);"3. Control Valve" 


12670 


LOCATE 22,1:PRINT TAB(26);"4. Sequence Start" 


12630 


LOCATE 17,54: PRINT ZDi^(ZT,l) 


12690 


LOCATE 19,46:PRINT ZD*(ZT,2) ;TAB(50) ; ZDt(ZT,3:) sTAB'S 


12700 


LOCATE 20,46: PRINT ZD*(ZT,4) ; TAB (50) ;ZD*(2T,5) ;TAB(5 


12710 


LOCATE 21, 53: PRINT ZD#(ZT,6) 


12720 


LOCATE 22,50: PRINT ZD* ( ZT , 7 ) ; TAB ( 54) ; "MIN" 


12730 


REM 


12740 


LOCATE 23,1 SPRINT TAB (73);" "; 


12750 


LOCATE 24,1: PRINT TAB(73);" "; 


12760 


LOCATE 23,5: INPUT "select number (5 = eKit)";AZ 


12770 


IF AZ <> 1 THEN 12350 


127S0 


LOCATE 23,5: INPUT "select either active(ACT) or not 


12735 


IF ZZ* = "" THEN 12350 


12790 


IF 11$ = "ACT" OR ZZ* = "act" THEN ZD*(ZT,1) = "ACT 


123.00 


IF ZZ$ = "ACT" OR ZZ* = "act" THEN 12350 


12310 


IF ZZ* = "NA" OR ZZiP = "na" THEN ZD*(ZT,1) = "NA" 


12320 


IF ZZ* = "NA" OR ZZf = "na" THEN 12350 


12S30 


LOCATE 24,5: PRINT "INCORRECT VALUE , TRY AGAIN"; 


12340 


GOTO 12730 


12B50 


IF AZ v> 2 THEN 13130 



"AR" 



acti ve(NA) 



PAGE 19 

12360 LOCATE 23,1: PRINT TAB (73);" ": 

12370 LOCATE 23 , 1 : INPUT"TIM IMG SEQUENCE 1 (ENTER TO DEFAULT) 0N/UFF";2: 

12330 IF ZZ$ = "" THEN 12930 

12890 IF ZZ* = "ON" OR lit = "or," THEN ZDi-''ZT,2) = "OM" 

12900 IF Hi- = "ON" OR ZZf = "on" THEN 12930 

12910 IF ZZ«: = "OFF" OR ZZ^ = "of-f" THEN ZD*(ZT,2) = "OFF" 

12920 IF 11$ = "OFF" OR ZZ* = "of-f" THEN 12930 

12925 GOTO 12360 

12930 LOCATE 23,63: INPUT "# OF NINS.";ZZf 

12940 IF lit- = "" THEN 13010 

12950 AQ = VALCZZ*) 

12960 IF AQ > AND AQ < 9999 THEN 2DJf(ZT,3) = ZZ* 

12970 IF AQ > AND AQ < 9999 THEN 13010 

12930 REM 

12990 LOCATE 23,63: PRINT TAB<7S);" "; 

13000 GOTO 12930 

13010 LOCATE 24,1: INPUT "TIMING SEQUENCE 2 (ENTER TO DEFAULT) ON/OFF"; 

13020 IF ZZ* = "" THEN 13070 

13030 IF ZZ-4: = "ON" OR ZZ* = "on" THEN ZD-^(ZT,4) = "ON" 

13040 IF ZZ* = "ON" OR ZZ:^ = "on" THEN 13070 

13042 IF ZZ* = "OFF" OR 11$ = "off" THEN ZD*(ZT,4) - "OFF" 

13044 IF II f = "OFF" OR ZZ$ = "off" THEN 13070 

13050 LOCATE 24,1: PRINT TAB(7S);" "; 

13060 GOTO 13010 

13070 LOCATE 23 , 63: INPUT "# OF MINS.";2Z* 

13075 IF ZZ* = "" THEN 13130 

13030 AQ = VAL(ZZ*) 

13090 IF AQ > AND AQ < 9999 THEN ZD:f(ZT,5) = ZZ4; 

13100 IF AQ > AND AQ < 9999 THEN 13130 

I 131 10 LOCATE 24,63: PRINT TAB (78) 5" "3 

13120 GOTO 13070 

13130 IF AZ 3 THEN 13210 

13140 LOCATE 23,1: PRINT TAB(73);" ": 

13150 LOCATE 23,1: INPUT "ENTER CONTROL VALVE (ENTER TO DEFAULT) "; ZZ* 

13160 IF ZZ* = "" THEN 13210 

13162 IF ASC(ZZ*) >47 AND ASC(ZZ*) < 91 THEN 13170 

13164 LOCATE 24,1: PRINT "USE CAPITAL LETTERS"; 

13166 GOTO 13140 

13170 IF LEN(ZZf) = 4 THEN ZDJr(ZT,6) = ZZ-? 

13130 IF LEN(ZZ*) = 4 THEN 13210 

13190 LOCATE 24,1: PR I NT "VALUE INCORRECT , TRY AGAIN" 

13200 GOTO 13140 

13210 IF AZ <> 4 THEN 13292 

13220 LOCATE 23,1: PRINT TAB(7'V^;" ": PRINT TAB(7^);" "; 

13230 LOCATE 23,1: INPUT" MINUTES FROM START UNTIL CONTROL BEGINS ";ZZ* 

13240 IF ZZ* = "" THEN 13292 

13250 AQ = VAL(ZZJ^) 

13260 IF AQ > AND AQ < 99'^9 THEN ZD-t(ZT,7) = ZZ* 

13270 IF AQ > AND AQ < 9999 THEN GOTO 132<=?2 

132S0 LOCATE 24,1: PR I NT "VALUE INCORRECT , TRY AGAIN"; 

13290 GOTO 13220 

13292 IF AZ < 1 OR AZ > 4 THEN 13300 

13294 FOR X = 15 TO 24 

13296 LOCATE X,l: PRINT TAB(73>;" "; 

k,132''3 NEXT X 



A6E 20 

3299 GOTO 12610 

33B0 IF AZ = 5 THEM 12220 

33 1 LOCATE 24,1: PR I NT " VALUE I MCORRECT , TRY AGA I M " 

3320 FOR X =^ 1 TO 10000; NEXT X 

5ZZi2 GOTO 12740 

3340 OPEN " D " , 1 H " 1 1 iTier . d at " 

3350 FOR X = 1 TO 10 

3360 FDR Y ---^ 1 TO 7 

3375 PR I NT # 1 , ZD* ( X , Y ) 

3330 NEXT Y 

3390 NEXT X 

3400 CLOSE 

3410 RETURN 



APPENDIX 2 
Assay Procedures and Instrumentation 



APPENDIX 2 - ASSAY PROCEDURES AND INSTRUMENTATION 

Enzyme Assays 

Three enzyme assay procedures were used to evaluate amylase activity of 
ATSH Enzyme preparations. All assay were in acetate buffer pH 4.0 at 35°C in 
a stationary water bath. 

1. Glucoamylase 

Glucoamylase catalyzes the release of single glucose molecules from the 
nonreducing end of starch or dextrin. Maltose is used as the substrate in a 
30-minute assay. Hydrolysis of maltose to glucose is measured on a Yellow 
Springs Instrument glucose analyzer. This instrument is specific for glucose; 
maltose is unreactive. 

2. Alpha Amylase 

Alpha amylase catalyzes the hydrolysis glucose linkages in the interior 
or starch chains producing shorter starch or dextrin chains. The assay uses 
amylopectin azure which is a blue dye bound to insoluble starch. Degradation 
of the starch molecule releases blue dye which is measured by a spectrophoto- 
meter (Bausch and Lorab Spectrines 21). Assay time is 15 minutes. 

3. Debranching 

Debranching activity catalyzes hydrolysis of alpha 1, 6 glucose linkages 
which occur at branch points in starch (alpha 1, 4 linkage form the straight 
chain portions of starch). Substrate for this assay was waxey or highly 
branched barley starch. The assay used a 2-hour time period and measured 
release of glucose. 



A-2.1 



Ethanol Assay 

Ethanol concentration In ATSH fermentation was determined by gas chroma- 
tography using a Varian Model 3700 gas chromatograph equipped with Portapack S 
column and flame ionization detector. Fermentation samples were centrifuged 
and diluted before being assayed. 

Glucose Assay 

Glucose in ATSH fermentations, enzyme assays and hydrolysis studies was 
determined using a Yellow Springs Instrument glucose analyzer. The enzymatic 
method used in this instrument is specific for glucose; maltose or higher raal- 
todextrins do not react. Samples were centrifuged and diluted as appropriate. 

Total Reducing Sugar 

Total reducing sugars were determined using a colorimetric procedure 
based on the reaction 3, 5 dinitrosalacylic acid (DNS) with reducing sugars. 
The reaction is nonspecific and measures all reducing sugar present in the 
sample. 



A-2.2 



APPENDIX 3 
Drawing and Schematics 



APPENDIX 4 
Mass and Energy Balance 



APPENDIX 4 - MASS AND ENERGY BALANCE - ASSUMPTIONS AND CALCULATIONS 



Mass Balance 



The mass balance is based on data from pilot plant runs. This data is an 
average value from the last three runs. 

1. Feedstock of 95% barley flakes and 5% barley straw by vreight. 

2. Feedstock contains an average of 10% moisture as received. 

3. Water and nutrients are added to bring the substrate to 51% 
moisture before cooking. 

4. Enzyme leaving the reactor averages 70% moisture by weight. 

5. 60% of the original dry substrate weight is recovered as dry enzyme. 

6. An inoculation dosage of 6.66 pounds of dried mold spore culture/ 
8,000 pounds total weight of substrate (wet) is required. 

7. 83.3% of the inlet oxygen is used over 72 hours. 

Laboratory fermentations of ATSH enzyme provide the starting point for the 
mass balance. It requires 0.375 pounds of ATSH enzyme from the pilot plant to 
yield one gallon of ethanol (ETOH). 

Therefore, 

20,000,000 gal/yr ETOH x .375 lbs ATSH/gal ETOH = 7,500,000 lbs ATSH/yr 

7,500,000 lbs ATSH/. 60 Recovery Rate = 12,500,000 lbs Dry Feedstock 

Adding H2O and nutrients gives a total wet substrate weight of 25,515,583 
Ibs/yr 

Inoculation requires additional mold culture: 

25,514,583 Ibs/yr x 6.66 lbs inoculant/8 ,000 lb. Subst = 21,240.9 lb 
mold spore culture 

The total enzyme production is 

7,500,000 Ibs/yr + 21,240.9 Ibs/yr = 7,521,240 Ibs/yr 

The total feedstock is now 7,521,240 Ibs/yr/. 60 or 12,535,400 Ibs/yr 

95% of the feedstock or 11,908,630 lbs are barley flakes; the remaining 
626,770 lbs are chopped barley straw. 



A-4.1 



Depending on the barley, approximately 35% of the total weight is lost in 
Che penrling/flaking operation. The total whole barley required for 
enzyme production is 11,908,630 lbs/yr/.65 or 18,320,970 Ibs/yr. 

The total substrate weight through the mixing and cooking operation and to the 
reactor is the feedstock weight plus water plus nutrients. 

12,535,400 Ibs/yr feedstock + 12,535,400 Ibs/yr H2O + 516,040 Ibs/yr Nut. = 
25,586,840 Ibs/yr 

The total weight leaving the reactor is equal to the weight entering the dryer. 
The material entering the dryer averages 70% moisture by weight. 

7,521,240 Ibs/yr Enzyme/. 30 = 25,070,800 Ibs/yr 

Feed to reactor is 25,586,842 Ibs/yr 

Wet enzyme from reactor is 25,070,800 Ibs/yr 

Weight loss in reactor is 516,042 Ibs/yr 

Wet enzyme to dryer is 25,070,800 Ibs/yr 
Dry enzyme from dryer is 7,521,240 Ibs/yr 
Weight loss in dryer is 17,549,560 Ibs/yr 

The weight loss in the reactor and the dryer is a result of converting hydrolyzed 
starch (glucose) to carbon dioxide (CO2) and water. The total starch loss is 
the total feedstock weight minus 10% moisture minus the dry enzyme weight. 

.90 (12,535,400) - 7,521,240 Ibs/yr = 3,760,621 Ibs/yr 

The chemical equation for the weight loss can be written as 

C^H]^206 + 6 O2 ""> 6 CO2 + 6 H2O 

For each mole of C6H]^206 lost, six moles of water (H2O) and six moles of carbon 
dioxide (CO2) are generated while six moles oxygen ( O9 ) are required. 

3,760,621 lbs loss = 20,892.34 moles C5Hi2^6 
180 lbs/lb mole C5H2^206 

20,892.34 moles C6H]^2^6 ^ 6 moles CO7 x 44 lbs CO9 = 5,515,567.2 lbs CO2 
mole C^H]^2^6 ^^ mole 

20,892.34 moles CfiHi206 ^ 6 moles O7 x 32 lbs O7 = 4,011,328 lbs O2 
mole C^Hj^2% ^^ mole 

20,892.34 moles Cf,ni20(, x 6 moles H9O x 18 lbs H?0 - 2,256,373 lbs H2O 
mole Cj^H2^2'^6 ^^ mole 

83.3% of the oxygen In the pilot plant was utilized so excess O2 is necessary. 

4,011,328 Ibs/yr/. 833 = 4,832,925 Ibs/yr 0-, 



A-4.2 



A mass balance is done around the reactor to determine the CO2 lost in the exit gas. 

Reactor In = 25,586,842 lbs subst + 4,832,925 lbs oxygen = 30,419,767 
Reactor Out = 25,070,800 + 821,598 lbs Oj = 25,892,398 Ibs/yr 
CO2 = 30,419,767 - 25,892,398 = 4,527,369 Ibs/yr 

Since 5,515,567 Ibs/yr of CO2 are generated and only 4,527,369 Ibs/yr are 
removed in the reactor, 988,198 lbs are absorbed in the wet enzyme and removed 
in the dryer. 

The water removed in the dryer is the sum of the following: 

H2O in feedstock = .10 (12,535,400) = 1,253,540 Ibs/yr 

H2O added = 12,535,400 Ibs/yr 

Metabolic H2O = 2,256,373 Ibs/yr 

Total 16,045,313 Ibs/yr 

In addition to the water and CO2 removed in the dryer, 516,048 Ibs/yr are lost 
between the dryer inlet and packaging. Part of this is a handling loss while 
the rest of it is volatiles and acids removed in the dryer. 

2. Energy Balance 

The theoretical energy requirements for an ATSH enzyme plant were calcu- ^ 
lated rather than typical process requirements. The theoretical values were 
given to equipment vendors to size equipment. For example, if drying requires 
1.67 X 109 Btu/yr theoretical heat, the actual energy input may vary from 
4.2 x lO^*^ Btu/yr to 1.11 x 10^^ Btu/yr. Heat recovery may vary from 50 to 78% 
of actual input. The theoretical values will remain constant while actual 
values vary according to type and size of equipment. 

Cooking Requirements 

Q = M Cp T 

where Q is the heat required, M is the mass, Cp is the heat capacity, 
and T is the temperature difference. 

Q = 25,586,843 Ibs/yr x 1 Btu/lb °F x (210 - 65°F) 
Q = 3.71 X 10^ Btu/yr 

Cooling Requirements 

Q = M Cp T 

Q = 25,586,843 Ibs/yr x 1 Btu/lb °F x (86 - 210°F) 

Q = 3.17 X 10^ Btu/yr 



A-4.3 



Drying Requirements 

17,549,560 Ibs/yr of material are removed in the dryer. 16,045,314 Ibs/yr 
of this is water vapor. Because the make-up of the remaining 1,504,246 
lbs is not precisely known, it is assumed to require the same amount of 
energy as the water fraction. 

The water is removed as H9Q vapor. The heat of vaporization is 965 Btu/lb. 

17,549,560 Ibs/yr x 965 Btu/lb = 1.69 x 10^0 Btu/yr 

Some cooling takes place in the dryer. The energy recovered from this is 
defined by the equation Q = M CP T 

Q = 7,521,240 Ibs/yr x 1 Btu/lb °F x (68 - 100°F) 
Q = 2.4 X 10^ Btu/yr 

The total theoretical energy requirement is 1.67 x lO^'^ Btu/yr. 



A-4.4 



APPENDIX 5 
Financial Analysis 



APPENDIX 5 - FINANCIAL ANALYSIS 

Financial analysis prepared by Anderson Zurmuehlen & Co., P. C. 
Note 

This analysis was prepared during August and September of 1986 and does 
not reflect changes in the costs of producing enzymes and by-products sold. 
The prices have changed due to process improvements resulting from development 
since the above period. Also, not reflected in this analysis is the reduction 
in by-product sales revenue due to a more recent determination of market price 
for that commodity. Some other expense categories have undergone minor adjust- 
ments. 

The accounting firm has not reviewed these changes and accepts no responsi- 
bility for the data shown in Table 3, page 57 of the report. 



A-5.1 



I PRSSJMPIARY DHAFT 

for Review and Discussk 
Subject to Change 



ATSH ENZYME PLANT 

PROJECTED FINANCIAL STATEMENTS 

FOR THE FIVE YEARS ENDING DECEMBER 31, 1992 



for Review ar.d DIscMssion 
Subject to Change 



CONTENTS 



PAGE 

REPORT OF CERTIFIED PUBLIC ACCOUNTANTS 

ON THE FINANCIAL STATEMENTS ■'■ 

FINANCIAL STATEMENTS 

Projected Balance Sheet 

Statement of Projected Results of Operation 
and Cash Flow - 5 Years 

Statement of Projected Results of Operations 
and Cash Flow - 1 Year 

Summary of Significant Projection Assxamptions 
and Accounting Policies 

Supplementary Schedule 10-17 



L^ 






ANDERSON ZURMUEHLEN 

Certifiea Public Accountants 

81 West Park Street P O Box 748, Butte, MT 59703 (406) 782-0451 



& 



CO, 

-1- 



P.C 



fcr Review and Discussjon 
Subject to Ch?^rj-,4i 



To the Officers 

Renewable Technologies, Inc. 

Butte, MT 59701 



4820 



We have compiled the accompanying projected balance sheet and the 
statement of projected results of operations and cash flows of the 
ATSH Enzyme Plant as of and for the five years ending December 31, 
1992 in accordance v/ith standards established by the American 
Institute of Certified Public Accountants. 

The accompanying projection and this report were prepared for 
Renewable Technologies, Inc. for the purpose of evaluating project 
financial feasibility and should not be used for any other pur- 
pose. 

A compilation is limited to presenting in the form of a projection 
information that is the representation of management and does not 
include evaluation of the support for the assumptions underlying 
the projection. We have not examined the projection and, accord- 
ingly, do not express an opinion or any other form of assurance on 
the accompanying statements or assumptions. Furthermore, even if 
financing is obtained and the plant becomes operational, there 
will usually be differences between the projected and actual 
results, because events and circumstances frequently do not occur 
as expected and those differences may be material. We have no 
responsibility to update this report for events and circumstances 
occurring after the date of this report. 



ANDERSON ZURMUEHLEN & CO. 
September 8, 1986 



P.C. 



Of FiCES Heierra. Biiimgs and Butte 

Mernoers o( American insitute of Certified Pubhc Accountants 



f^ember of Associated Regional Accounting Firms 
Member of Private Companies Practice Section of AlCPA 



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5 5^15 s i I ii 



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675,345 $4,223,750 
125,742 $1,144,553 

549,533. $3,079,197. 

$74,312 $76,983 
326,044. $339,52B- 
178,326 $164,570 


582,687. $603,081 
966,896 • $2,476,116. 
206,9461 ($153,8001 
759,950 $2,317,316 
584,431) ($774,135)- 
173,319. $1,541,180. 

797,468 $2,976,298 
609,341 $4,159,962 

407,010 $7,136,280 
710,005 $1,746,527 
534,631 $776,135 
134,075 1134,075 
430,711 $2,658,737. 
976,298 $4,477,543 




107j714 $3 
093,023 $1 

014,493- $2 

$73,731 
314,953- 
172,298 

562,982 

451,711- $1 

262,491)- ( 

189,220 $1 

392,583) ( 

796,437. $1 

926,448 $1 
054,082 $3 

982,529 $5 

458,203 $1 

392,583 

134,075 

164,661 $2 

797,668 $2 


CO 


" ~ i ** ! 1 r r Mi 1 ^ i'^ i" r 1 "" ii 


ATSH EN7YI1E PLANT 

STATENENT Of PROJECTED RESULTS OF OPERATIONS AND CASH FLO 

FOR THE FIVE YEARS ENDIN6 DECEKBER 31, 1992 

; CONSTRUCTION 1 PERIOD 1 1 PERIOD 2 
i PHASE : 1 


NET SALES $1,640,672 $2,646,46C 
Less: COST Of 6000S SOLD «804,109 $1,046,8! 

MOSSPROflT $0 $1,034,563 $I,599,64< 

Less: Sales Expenses $68,629 $71,23£ 
Seneral I Adiinistrative Expeoscs $295,879- $304,23 
Plant Overhead Costs $140,842 $144,47 


$0 $525,550 $543,94 
OPERATING PROFIT $0 $511,013 . $1,055,70 
Less: Other Expenses ($419,504). (1327,76 
NET PROFIT BEFOflE TAIES $0 $91,508. $727,93 
Incoie Tax Provision $0 ($19,341) ($235,74 
NET INCOK (LOSS) $0 $72,148. $492,19 

CASK BALANCE (Openinj) $242,500' $347,80 

Plus RECEIPTS: Receivable Collections $1,424,640 $2,552,74 
Equity Financing $1,800,000 
Bank Loan Proceeds $500,000 

TOTAL $2,300,000. $1,849,140 $2,900,56 

Less DISBURSEHENTS: Trade Payables $1,347,898- $1,602,29 
Fixed Asset Additions $2,057,500 

IncoM Taxes $19,341 $235,74 
Dividends Qr Nithdramls 
flank Loan Repayient $134,075 $134,07 

TOTAL $2,057,500 $1,521,334, $1,972,12 

CASH BALANCE (Closing) $242,500. $347,804 $928,44 




^ 



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is i i s n a i 

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fe va ill '- 3 II - « 

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ea S f- f- I in 5 



i - 



PRELIWINARY DRA 

for fWview and Di3c:if 

...JBubiect to Chan 

-5- 



ATSH ENZ'/TIE PLANT 

SUMMARY OF SIGNIFICANT PROJECTION ASSUMPTIONS AND ACCOUNTING POLICIES 

FOR THE FIVE YEARS ENDING DECe-IBER 31, 1992 

This financial projection presents to the best of management's 
knowledge and belief the Plant's expected financial position and 
results of operations and cash flow for the projection period. 
Accordingly, the projection reflects its judgement as of September 1, 
1986, the date of this projection, of the expected conditions and its 
expected course of action. The assumptions disclosed herein are those 
that management believes are significant to the projection. There will 
usually be differences between the projected and actual results because 
events and circumstances frequently do not occur as expected and those 
differences may be material. 

NOTE 1. FORMATION OF THE PLANT AND SIGNIFICANT ACCOUNTING POLICIES 

Description of the Plant: 

The proposed ATSH Enzymes Plant is a manufacturing concern 
capable of producing an enzyme which will be used to produce 
ethanol. Construction of the plant to be located on three 
acres of land in an as yet undetermined location, is antici- 
pated to begin on July 1, 1987 and be completed by January 1, 
1988. Equity financing is proposed to be obtained from a 
venture capital company concern. The proceeds from the 
venture capital company, together with anticipated bank 
financing, are expected to provide the necessary funding 
requirements for the project. 

Valuation of Trade Receivables: 

Trade receivables are stated at net amounts with no allowance 
for doubtful accounts. An allowance for doubtful accounts is 
not considered necessary because it is considered immaterial 
for purposes of this projection. 

Inventories: 

The Company does not distinguish between raw materials, work 
in process and finished goods inventories. Anything consti- 
tuting inventory to the Company is stated at projected costs 
to include the cost of raw materials, direct labor and other 
manufacturing costs. 

Depreciation: 

Depreciation has been provided for under the guidelines of 
the 1986 Tax Reform Act. It should be noted that the tax 
bill is not in final form and, therefore, subject to 
revision. The 200% declining balance method of depreciation 
was used for all manufacturing equipment with estimated 
useful lives of ten years. The building depreciation was 
provided under the straight-line method with an estimated 
useful life of 31 1/2 years. 



PREL1MJ?\!ARY CRAFT 

for Review and Discussion 

Subject to Change 

-6- 



ATSH ENZYME PLANT 

SUMMARY OF SIO^IFICANT PROJECTION ASSUMPTIONS AND ACCOUNTING POLICIES 

FOR THE FIVE YEARS ENDING DECEMBER 31, 1992 



NOTE 1. (CONT'D) 

Research and Development: 

Research and development costs are charged to operations as 

incurred. 

Income Taxes: 

The Company has provided for income taxes under the 
guidelines of the 1986 Tax Reform Act, As mentioned above, 
this legislation is not in final form and, therefore, subject 
to revision. Income tax expense was computed using the 
following rates: 

$ -0- - $50,000 15% 
$50,000 - $75,000 25% 
$75,000 - Over 34% 

Income tax credits are accounted for by the flow through 
method which recognized the credits as a reduction to income 
tax expense in the year utilized. 

NOTE 2. SALES 

Sales are based on management's estimates of the number of 
pounds of enzymes sold each year at the expected selling 
price per pound which is illustrated as follows: 

Sales Period 1 Period 2 Period 3 Period 4 Period 5 

Enzyme Sales: 
Lbs./Yr. 6,297,200 8,721,622 9,048,919 9,093,104 9,099,069 
Price/Lbs. ^ .Jl .11 ^ A3 

Total $1,637,272 $2,354,838 $2,805,165 $3,364,449 $3,912,600 



By-Product Sales: 

TonsAr. 1,695 2,348 2,436 2,448 2,450 

Price/Ton $120.00 $124.20 $124.20 $127.00 $127.00 

Total $203,400 $291,622 $302,551 $310,896 $311,150 

Total Sales $1,840,672 $2,646,460 $3,107,716 $3,675,345 $4,223,750 



^ # 



PRELiai?NARY CRAFT 
for Review ar^o* Dior::'j:«sfc 
Subject to_ Change 

ATSH ENZYI1E PLANT 

SUMMARY OF SIGNIFICANT PROJECTION ASSUMPTIONS AND ACCOUNTING POLICIES 

FOR THE FIVE YEARS ENDING DECEMBER 31, 1986 



NOTE 2. (CONT'D) 

The market demand for the product is assiomed to be strong due 
to the refining efficiencies of the enzyme in comparison to 
the refining techniques currently on the market. One impor- 
tant assumption made by management which accounts for the 
annual increases in sales concerns the improved dose rate of 
its product. The dose rate is the amount of enzyme needed to 
produce one gallon of ethanol. The projections assume that 
the plant will be at full production capacity by the end of 
year two but sales will continue to increase due to the 
improved dose rate of the enzyme and the corresponding 
increase in the price which the Company will be able to 
demand for the enzyme. 



NOTE 3. COST OF SALES 

Raw Materials. 

Raw materials used by the Company are expected to be readily 
available and the Company has used historical information 
relative to the actual cash price of barley to arrive at its 
projected price per bushel. The price of barley, the major 
raw material in the Company's product, has fluctuated 
significantly over the last ten years. Due to this 
uncertainty, the realization of this projection is 
particularly sensitive to the actual cash price of barley and 
any variation from the projected price would significantly 
affect the projected net income. The raw materials prices 
used for purposes of this projection are as follows: 



1988 


$2.15/Bushel 


1989 


$2.20 


1990 


$2.20 


1991 


$2.25 


1992 


$2.25 



Direct Labor. 

The Company has projected a labor force of eleven employees 
to operate the plant. Compensation was arrived at by 
referring to industry standards of manufacturing concerns ir 
a similar industry. Total labor costs include fringe 
benefits calculated at 33% of base salaries. The Company 
does not expect to be dealing with labor union contracts. ] 
the event that they do become involved with a labor union, 
labor costs may change significantly. The outcome of the 
projection is significantly sensitive to variances in such 
labor costs. 



PRELir/lfNARY CRAFT 

for Review and LTi.'jc^jssior^ 

Subject to Changs 



ATSH ENZYME PLANT 

SUMMARY OF SIGNIFICANT PROJECTION ASSUMPTIONS AND ACCOUNTING POLICIES 

FOR THE FIVE YEARS ENDING DECEMBER 31, 1992 



NOTE 4, 



BANK BORROWINGS AND INTEREST EXPENSES 



The forecast assumes that a certain amount of financing for 
the project will be obtained from a bank. The Company used 
an interest rate of 11% annually to calculate interest 
expense. This projection was based on quotes from local 
banks. The interest calculation was based on a five year 
term and no allowance was made for the floating nature of the 
rate which would probably be the case with a loan of this 
type. 



NOTE 5. 



SALES EXPENSES, GENERAL AND ADMINISTRATIVE AND PLANT 
OVERHEAD COSTS 



Selling expenses include salaries, commission and traveling 
expenses for salesmen as well as shipping costs, costs for 
containers and advertising expenses. General and 
administrative expenses include salaries and wages for 
administrative secretaries, accountants and similar workers 
as well as office supplies, equipment, outside communications 
and other administrative activities. Plant overhead costs 
include all other costs not directly related to the 
production process. These costs include safety services, 
cafeteria facilities, janitor services, employment offices, 
shops and warehouses. The majority of the at»ove costs were 
projected based on industry standards while some of these 
costs were based on an independent estimation. 



NOTE 6. OTHER EXPENSES 



Other expenses include all non-operating expenses such as 
interest and depreciation. The calculation and projection of 
these expenses has been addressed previously. 



NOTE 7. OTHER ASSUMPTIONS 



In determining inventory levels, trade accounts payable and 
receivables management utilized standards for chemical 
manufacturing conpanies. A summary of the ratios used 
follows: 



Trade Receivables 
Trade Payables 
Inventory 



Turnover 

8.6 Times 
9.2 Times 
6.4 Times 



Number of 
Days 

42 
40 
57 



« 4 



P9.EUTV.:i,'^ny DRAFT 
for ftevjev,' jrd Disciissio 
Subject to Change 



ATSH ENZYI1E PLANT 

SUMMARY OF SIGNIFICANT PROJECTION ASSUMPTIONS AND ACCOUNTING POLICIES 

FOR THE FIVE YEARS ENDING DECEMBER 31, 1992 



NOTE 7. (CONT'D) 

In reviewing other key financial ratios, management is aware 
of existing discrepancies. The reason for these 
discrepancies is that in certain instances management chose 
to use an independent estimation of certain costs rather than 
refer to industry standards. Their basis for these decisions 
is due to the fact that the technology is new and, therefore, 
has no industry track record which can be referred to. 

One final noteworthy assumption made by management concerns 
the use of an inflation factor at 3.5% to determine cost 
increases from year to year. 



ENZYME SALES: 

LSS./VR, 



/c 

for Reviev; n: Dir'^-ission 

-Subject to Change ^ 



FOR THE FIvE VEARE ENDEE CEZEMBER 31, lYFc 



PERIOD ! 


--FIjD e 


[NFLATION RATE 


3.S' 


6,E?7,e00 


8,"5!-i?' 


$0.26 


?0.E' 



SY-'^'RQL'l'CT SALES: 

'ONS/YR. !!6?5 3,3^3 l,h 

PRICE'TON IIEO.OO tiE^.EO l!S-. 

TOTAL ?203)4Q0' SE'liicE. 1302, S 

TOTAL SALES $1,3^0,672- %^^rMh^^hfi■ $3-107,' 



PRELIT":: 'r:r draft 

for Reviev^ a id Discussion 
Subject to Change 



AT3H ENZYHE PLANT 

SUFPLEKENTAL SCHEDULE 

FOR THE FIVE ''EARS ENDED DECEfiBER 31 1 !ttc 



EN2y?*E SY-f'RQDUCT 



3E:-INN!N6 inventgry 








FRGDUCTION FERIQQ 1 


7,280,000 


1.960 


^QTAL AVAILABLE 


7,230,000 


!i'60 


INVEHTCRV HELD 'IS.SX) 


! 032, 300 ^ 


■.2'zZ'r 


UNITS SOLD PERIOD ! 


6,297,200. 


li-'-'S. 


?E=IN'N!M5 INVE«JTGRY 


533, eoo 


265 


PRCDUCTIOM PERIOD 2 


?, 100,000 


2,^50 


TOTAL AVAILABLE 


10,032,300 


2,715- 


INVENTORY HELD !13.!r.! 


!!,3il,173?- 


!366:i- 


UNITS SOLD PERICD £ 


3,721,i£2. 


2,343. 


SESIN'N'INS INVENTORY 


•,361,173 


346 


FRQDUCTIGN PERIOD 3 


9,100,000 


2,^:0 


TOTAL AVAILABLE 


10,^61,173 


2,316 


INVENTORY HELD (13.5!;) 


(!,Vi2,259). 


!330! 


UNITS SOLD PERIOD 3 


9,0^3, 91'. 


2,ij36 


SESINHIN5 INVENTORY 


l,il2,2S9 


330 


PPCDUCTION PERIOD «t 


9,100,000 


2,i50 


TOTAL AVAILABLE 


I0,S12,2!9 


2,330 


INVENTORY HELD !!3.5X) 


(l,i(19,155)- 


(382) 


UNITS SOLD PERIOD ^ 


9,093, lO^t. 


2,-.i3. 


SESINNING INVENTORY 


1,^19,155 


•J03 


PRODUCTION PERIOD 5 


9,100,000 


2,r50 


TOTAL AVAILABLE 


10,519, IS5 


2,832 


INVENTORY HELD !13.5r>) 


( 1,^,20,036 )• 


(382 J 


UNITS SOLD PERIOD 5 


9,099,069 


2,«0. 




::::i:rsri::::: 


=============== 


3E5[;iH!NS INVENTORY 


1,«0,036 


382 


PRODUCTION PERIOD o 


9,100,000 


2,«0 



TOTAL AVAILABLE 
INVENTOR" HELD (13.5"! 



■■jj-q cci ri :'c;: 



5EEINNING INVENTORY 
■RODUCTION -ERIQD 3 



TOTAL AVAILABLE 
iVENTQRY HELD (13.5X1 



ONITS SOLD PERIOD 3 



BEGINNING INVENTORY 
FRODIJCTION PERIOD 9 



TOTAL AVAILABLE 
INVENTORY HELD '13.5") 



UNITS SOLD PERIOD 9 



ATSH ENZY"E PL-NT 



--Q J-^r r-TiT 



INVENTORY HELD '13.5"; (li^SOiElE; 

!'NIT- 3"LD PEPiOD 4 ?, 099137^ 






10,520,212 


2 i 332 


(1,420,22?! 


(332!- 


0,;V3Q,Oa3. 


3,t5::i 


1,^20,239- 


339 


9, !n(i,|)!)n 


3ir50 


10,520,229 


2,332 


(l,it20,2315 


!3S2! 


a.QOO.Ong^ 


2, ■--_:] 







1,420,231 


332 


9,100,000 


2,450 


10,520,231 


2,832 


(l,420i2311 


(332! 


?i 100,000 


2; 450 



PRELIMINARY DRAFT ^^ 
for Review arrd Discussion 
Subject to Change 



FRICE'PL'SHELL 



iOTSL 



PRELIMIi>iARY DRAFT 

for Review ad Diccussion 

S^lbiect to Change 

.ANT 



:UFrL£^ENT;;L SCHEDULE 
■OR THE FIVE YEARS ENDED DECEMBER 31, !«'E 



:-LIS ^ErlOj ! FE.-.IGD z 



lA'ERIALS: 

2'JSHELLS/YR. 112,22": E?*,ci7 i^l-ioT 5?'. -A? 



:3 ;rt 






INFLATION RATE I.:-;" 

PERIOD 1 PERIOD 2 ^ERIGD 2 PERIOD - PERIOD " 

OTHER «'FG. COSTS: 

PHR. i ^TIL. -25,072 $25,^50 $26,3S8 527,7^5 122=7"' 

fl.AINT. !: REPAIRS 5=3,700 ;71i!05 173,593 $76, 16= :7sic25 

OPER. SUPPLIES ?10,30; 5!;,iii l!i,03? ^-U^i^ HlisES 

LAB SERVICE: ?:3,152 555,012 I5sj?33 553,':'! Sioi^'3 

RQVfiLTIES 50 JO $0 IC '" =0 

OTHER PURCH. INPUTS I5,0?i 55,27^ 55,^59 55,^50 55,3^3 

"TAL Sli2|325. 1163,006 $172, :37. $:7?,?7S $136, £75. 



DIRECT LABOR: 



// 



ATSH ENZYME PLANT 
SUPFLEHENTAL SCHEDULE 
THE FIVE VEARS ENDED DECE.1BER 31, 1992 



FRINGE 
§ 331 



TOTAL 
OF E!1P. J/YEAR 



PLANT (1ANAGER 


N/A 


SALARY 


J35,500 


$11,715 


$47,215 


1 


$47,215 


PLANT FOREMAN 


N/A 


SALARY 


J26,000 


$8,580 


$34,580 


1 


$34,580 


PRODUCTION TECH. 


2,0B0 


J9.50 


J19,760 


$6,521 


$24,281 


7 


$183, '64 








$81,240 


$24,814 


$108,074 


9 


$245,741 






INFLATION RATE 


3.501 










YEAR 2 








FRINGE 






TOTAL 




» OF HOURS 


J/HOUR 


$/YEflR 


« 33Z 


$/YEAR 


» OF ENP. 


$/YEAR 


PLANT MANAGER 


N/A 


SALARY 


$34,743 


$12,125 


$48,348 


1 


$43,843 


PLANT FOREMAN 


N/A 


SALARY 


$24,910 


$8,880 


$35,790 


1 


$35,790 


PRODUCTION TECH. 


2,080 


$9.83 


$20,452 


$6,749 


$27,201 


7 


$190,404 



$111,853 



YEAR 3 


1 OF HOURS 


»/HOUR 


$/YEAR 


FRINGE 
a 331 


$/YEAR 


1 OF EMP. 


TOTAL 
$/YtAR 


PLANT NANAGER 
PLANT FOREMAN 
PRODUCTION TECH. 


N/A 
N/A 
2,080 


SALARY 
SALARY 
$10.18 


$38,028 
$27,852 
$21,147 


$12,549 
$9,191 
$4,935 


$50,578 
$37,043 
$28,153 


1 
1 
7 


$50,:73 
$37,043 

mi 








$87,048 


$28,724 


$115,773 


9 


$284,689 


YEAR 4 


1 OF HOURS 


$/HOUR 


$/YEAR 


FRINGE 
« 331 


$/YEAR 


t OF EMP. 


TOTAL 
$/YEAR 


PLANT MANAGER 
PLANT FOREMAN 
PRODUCTION TECH. 


N/A 
N/A 
2,080 


SALARY 
SALARY 
$10.53 


$39,359 
$23,827 
$21,908 


$12,989 
$9,513 
$7,230 


$52,348 
$38,339 
$29,138 


1 
1 
7 


$52,348 
$38,339 
$203,944 








$90,094 


$29,731 


$119,324 


9 


$294,654 


YEAR 5 


1 OF HOURS 


S/HOUR 


$/YEAR 


FRINGE 
8 331 


$/YEAR 


1 OF EMP. 


TOTAL 
$/YEAR 


PLANT MANAGER 
PLANT FOREMAN 
PRODUCTION TECH. 


N/A 
N/A 
2,080 


SALARY 
SALARY 
$10.90 


$40,737 
$29,834 
$22,475 


$13,443 
$9,844 
$7,483 


$54,180 
$39,481 
$30,158 




$54,180 
$39,681 
$211,105 



$93,248 



PRELifT^MARY DRAFT 

for Review a,.d Discussion 

Subject to Change 



ATSH ENZYKE PLANT 
:i;rPLE?!ENTAL =CHEDULE 
FGF: THE -'Ml YEARS EN'rED DECEr:-E?: ::, 



^VENTCRY 



:"• INV- '-') ^iES.C-i ;::;,:c: srOJi: 51'", 90^'^ 

^2D: PURCHASE: f501,i=6 1:i!,i67 S:-!.-? $656."; =^5V"' 

DIRECT LABGR 1265,76! SE75,06E 5E:-r:E' ?2'i,6:^ pni.^H 

CTHEP f1''5 E^P ti62,225 $!iS.0O6 3:":.:3' s:"^-:": r.2b.fl 

530CS AVAILABLE 592?, 752, ;i,21v,3-E i;.E::,S05 SS--:,-'- r. •:-,-- 
LESS:CCST OF SOGCS 

SOLD JSO'iiiO? $!i!'i46!3;- HiO'- 3 ■''•?' t',-:"^.-.; .-; :;.:. -r-, 

END.'NS INVENTORY JlEJ-i'-E r-","-"- «'^a "S'^ j;-^ 5;-;-s r--:- n^: 






''ERIGD ! -ERIGD : =E?!C: Z or^-nr. _ :c:!nr. 



P^l^"^?ll $67, 70' $5!,02! 5!!}!,55i 1112.305 siii.-;" 

INTEREST $66,371 543, 05^ --32,522 "i'^ZrZ "t^'^-a 



tm,m. $134,075. 5134,075. 



PREL!r/:3r4ARY DRAFT 

for Review ard Discussion 

Subject to Change 



/i 



N. 


. AD!1IN.: 








SEN. i ADMIN. 


$??,;£' 


ISE,5SC 




INSURANCE 


$65,000 


$67,E7S 




R&D 


jl^Sjijf' 


*!03,330 




TA'ES - FRGPERTY 


t i -.'■.:'• n 


i^'zV) 









$liO,3^E 



INTEREST INC. 








INTEREST EXP. 


$66,371 


5^3,05^ 


$32,5SE 


DEPRECIATION 


$353,133 


t-ESij-^OS 


$239, ?69 



til5,50!j 



SUPPLEMENTAL SCHESULE 
-I'll 'EARS ENDED DECEASE?: 



PRELIMINARY DRAFT 

for Review ad Discussion 

Subject to Change 



;•: " 


3LLECTICN 


A/R 


SEi:. :AL. 


ADD: 


SALES 


LE:S 


: A/" E!!D. :AL. 


CASH 


•uLL-CTIONS 



t-0 


Sc!^,03E 


;30"<7E3 


:3ili3i;E 


I'' 


$1,SW,67H 


$E,6^i:,'t60 


$3,107<714 


!3,o7S,3'r: 


5-.E33i7!0 


S!,3w,i7E 


!2,StCNi?l 


s3,4!;,iii 


?i. 036.707 


;^. :!!,:•; 


5E!i,03£ 


:-30',"ES 


=3ili3iZ 


=-3-,3i: 


^--lii:- 



!!,cEi,i-0 



ciyjBi c: n'ci;i!5crMC>'Tr: 



A/P BES. BAL. 

ADD: PURCHASES 



.ESS: A/P END. SAL. 



-AS.H DISBURSED 



$0 


53^,^03 


$113,754 


$113,307 


$1£E,3=5 


$!,^5S,302 


$l,6E3,c30 


51,r43,E26 


$1,713,^64 


;■ i7!0,570 


ti, -5,302 


n, 714,033 


$1,7'^'^,010 


JliSSEiS"^"' 


$1. 3-3, ^35 


$37,403 


$113,73;, 


$113,307 


$132,36: 


::E4,-v3 


$l,3i7,8?3. 


$1,602,2??. 

:r ============= 


$1,653,203. 

============== : 


$1,710,005. 

============== =: 


$l,^i?,!2^ 



"^ 



?