1. Biohydrogen Production by Thermotoga
neapolitana using Peach Waste Substrate
Alex Ryan, Mike Perkins, Zach Gilstrap, Zack Montgomery
http://america.pink/thermotoga-neapolitana_4422979.html
2. Introduction: Problem
Recognition of Problem
Growing population in the world exerts high energy demand unable to be met sustainably
Excessive use of fossil fuels causing impending energy crisis and global warming
Agricultural waste presents untapped potential in meeting energy needs
Definition of Problem
Musser Farm grows peaches strictly for research creating substantial peach waste (133,000 lbs/yr)
Peach waste usually thrown into waste ponds, soon may have to pay to dispose of waste
Peach waste has potential for biohydrogen gas production via fermentation using Thermatoga neapolitana
Biohydrogen gas potential energy source for hydrogen fuel cell electricity production
3. Introduction: Goals
Project Goals
Design a process to consume 100% of Musser Farm peach waste via fermentation pathways
Successfully capture and utilize biohydrogen product for energy needs
Potential energy for hydrogen fuel cell
Characterize effluent waste and determine sustainable alternative uses
Generate an overall positive annual worth
Biological Goals: Culture Thermatoga neapolitana and allow fermentation of peach waste from Musser Farm to
generate biohydrogen gas. Find alternate uses and create bioproducts from the remaining peach waste not converted to
biohydrogen gas in the effluent.
Structural Goals: Design a storage system and reactor able to hold and process 100% of the Musser Farm waste.
Mechanical Goals: Design a reactor to process Musser Farm peach waste and produce biohydrogen gas. Design a
heating process to maintain an optimal temperature of 77℃ for bacterial growth and fermentation at an appropriate mixing
speed. Design pumping system between unit operation.
4. Introduction: Constraints
Skills: Skills in programs such as SuperPro Designer and STELLA will be a major factor in the design
specifications of the peach waste system. Required to have skillful labor to operate the process.
Budgetary: The bioprocess design has to prove to be an economically viable alternative
Space: The reactor system has to be able to fit on site at the Cherry Crossing Facility or Musser Farm.
Logistics: The reactor must be able to be assembled on site or easily transported from the build site to the final
location. Reactor inlet and outlet components must be able to be stored and separated.
Time: The design must be completed in the allotted time period of one semester. Peach growing season limited
to four months (120 days), making storage a necessity. Goal is to design a reaction process capable of reacting
all peach waste within growing season.
5. Introduction: Considerations
Safety: The design should be simple and safe to use by all parties who will operate it.
Ethical: All ethical concerns for this project would be summarized in ecological concerns.
Ecological: The processes carried out by the reactor, and any later processes using products of the
reactor, should have as low of an ecological impact as possible. The main goal of the design is to reduce
the amount of product being wasted and to find an alternate use for the effluent flow.
Ultimate Use: To design an alternative energy process which utilizes agricultural waste. This process will
convert a waste into energy, helping the facility lower the overall non-renewable energy usage and lower
the whole facility’s carbon footprint.
6. Introduction: Questions
User (Cherry Crossing or Musser Farm Operator)
How long does the operation take?
What do I do with the reactor waste?
What components are added to the reactor?
7. Introduction: Questions
Client (Cherry Crossing or Musser Farm Manager)
How much will the implementation cost?
How much money will it save us?
How much energy can this process create on an annual basis?
8. Introduction: Questions
Designer (Us)
How much waste is there needed to be processed?
How pure does the gas stream need to be?
How long does the reactor need to be in operation?
9. Literature Review: Design Options
Anaerobic Digester: Methane Gas Production
Requires: lipids, carbohydrates (sugars), and protein
Lack of lipids and proteins in peach waste
Mixed stream with wastewater
Ethanol Production via Fermentation
High sugar content
Freely available disaccharides (sucrose)
Low ethanol yield of 25.68 g ethanol/ g dry peach
Hydrogen Gas via Fermentation
High sugar content
Freely available monomer sugars
High Hydrogen yields ≃32 mmol H2/L of media
10. Literature Review: Governing Equations
Overall reaction for fermentation of glucose via the EM pathway
C6H12O6 + 2H2O + 4ADP → 2CH3COO- + 2H+ + 2CO2 + 4H2 + 4ATP (Drapcho et al., 2008)
C6H12O6 + 0.5H2O + 0.34NH3 → 1.72CH1.8O0.5N0.2 + 1.25CH3COOH+ + 2.5H2 + 1.25CO2 (Yu, 2007)
Characteristics of Thermatoga neapolitana
Optimal pH: 7.0
Optimal Temperature: 77 deg C
Maximum Specific Growth Rate (μmax): 0.94 hr-1 (Drapcho et al., 2008)
Half Saturation Constant (Ks): 0.57 g/L
Product Yield (YP/S): 0.0278 g P/g S
Biomass Yield (YX/S): 0.235 g XB/g S
Product Formation Ratio (kpg): 0.118 g P/g XB
11. Literature Review: Governing Equations Cont.
Monod equation
Batch Reactor
Continuously Stirred Tank Reactor
Drapcho, C. M., Nhuan, N. P., & Walker, T. H. (2008). Biofuels engineering process
technology. New York: McGraw Hill.
Drapcho, C. M., Nhuan, N. P., & Walker, T. H. (2008). Biofuels engineering process
technology. New York: McGraw Hill.
12. Literature Review: Governing Equations Cont.
Heat Transport Equations:
● Heat required in temperature
change for a batch system
● Heat flux due to irradiance
● Batch solar heater energy balance
● Heat transfer rate in a single
hollow cylinder with convection at
surfaces
Incropera, Frank P., and David P. DeWitt. Fundamentals of Heat and Mass Transfer. New York: J. Wiley, 2002. Print.
13. Literature Review: Options to Address Problem
Reactor Options:
Batch Reactor
Multiple Reactors in series/parallel
Continuously Stirred Tank Reactor
Multiple Reactors in series/parallel
Recirculation Flow
Biohydrogen Use:
Gasifier
Hydrogen Fuel Cell
Effluent Use:
Vinegar
Composting
Recirculation
Enzymes
Reactor Component Options:
Nitrogen Source
Canola Meal
Soybean Meal
Trypticase
Carbohydrate Source
Peach Slurry
Water Source
DI Water
Peach Wastewater
pH Controller
Salt Mixture
Water Heating Options:
Preheating Before Reaction
Heating Throughout Reaction
14. Literature Review: Hard Data
Drapcho, C. M., Nhuan, N. P., & Walker, T. H. (2008). Biofuels engineering
process technology. New York: McGraw Hill.
15. Literature Review: Hard Data
Hill, Louis, "Improvements in the process of biohydrogen production by
Thermotoga neapolitana" (2013). All Theses. Paper 1796
16. Literature Review: Hard Data
Drapcho, C. M., Nhuan, N. P., & Walker, T. H. (2008). Biofuels engineering
process technology. New York: McGraw Hill.
17. Literature Review: Hard Data
Drapcho, C. M., Nhuan, N. P., & Walker, T. H. (2008). Biofuels engineering
process technology. New York: McGraw Hill.
Peak H2 Produced
(mmol H2 gas/L-hr)
C. saccharolyticus: 8.4
T. neapolitana: 4.5
18. Literature Review: Past Experiences & Heuristics
Fermentation of a feedstock in batch reactors and CSTRs
Modeling growth of microorganisms
Experience with Biomass and SuperPro Designer
Knowledge from Creative Inquiry
Utilizing heat transfer equations theoretically and experimentally
19. Literature Review: Preliminary Data Collection
Musser Farm Visit:
Peaches are grown for research and root
systems
Research Peaches = Majority Red Haven
Peaches
Peaches for Seeds = Guardian Peaches
Approximately 45 acres dedicated to Red
Haven peaches (308,000 peaches)
Waste formed in two ways
Stone separation from guardians
Peach discarding after research
20. Design Methodology: Preliminary Data
CollectionPeach Pit Separation
- Stone separation involves mixing/filtration
process to separate cull from seed
- At Musser Farm, stones are collected and
cull is released into collection pit
- Use a similar process to remove peach
seeds and prepare culls into slurry
Figure 1. Visual Representation of Peach Pitter
21. Literature Review: Preliminary Data Collection
Musser Farm Visit:
Peach pits separated and collected while culls
discarded in open pit
Requires periodic dredging to remove buildup
Potential for high nutrient reclamation
Goal of this project is to determine more
economically efficient alternatives to avoid
fines
22. Design Methodology: Analysis of Information
Total Peach Waste ≅ 60,000 kg (66 tons)
Total Glucose per Day ≅ 40 kg ≅ 1.7 kg/h
Total Batch Reactor Volume ≅ 5000 L
Working: 4000 L
Headspace: 1000 L
Total CSTR Volume ≅ 1500 L
Working: 1200 L
Headspace: 300 L
Table 1. Determination of Reactor Volumes
23. Design Methodology: Analysis of Information
Inlet
Concentration
[g/L]
Batch
[kg/day]
CSTR Flow Rate
[kg/h]
Peach Slurry 125 485 20.2
Glucose 10.0 38.9 1.70
Canola Meal 5.50 21.4 0.90
Salt Mixture 13.2 51.5 2.15
Water 865 3370 140
Biomass 0.01 0.04 0.002
Table 2. Determination of Reactor Components
24. Alternative Design Options
Methods of Heating:
● Electric generator
○ Cost, practicality of
implementation
● Solar energy
○ Cost, shade covering
● Exhaust from gasifier
○ Amount of heat generation
Process Step:
CSTR
Optimized growth
High peach waste requirement
Smaller Volume
Batch
Non-optimized growth
Variable peach waste
requirement
Higher volume
25. Design Methodology: Synthesis of Batch
Reactor
Predicted Time Until Full
Substrate Utilization:
t ≅ 7.5 hours
Predicted Hydrogen
Concentration at Full
Substrate Utilization:
[H2] ≅ 0.280 g/L (1.1 kg/d)
*Growth Variables Listed in
Literature Review Section*
XBi = 0.01 g/L
Si = 10 g/L
Figure 4. SuperPro Fermentation Batch Reactor Design
26. Design Methodology: Synthesis of Batch
Reactor
Figure 3. Predicted BIOMASS Batch Reaction Outcome
Predicted Time Until Full
Substrate Utilization:
t ≅ 7 hours
Predicted Hydrogen
Concentration at Full
Substrate Utilization:
[H2] ≅ 0.278 g/L
*Growth Variables Listed in
Literature Review Section*
XBi = 0.01 g/L
Si = 10 g/L
29. Design Methodology: Synthesis of CSTR
Ηydraulic Retention Time:
τ ≅ 7.5 hours
Predicted Rate of Hydrogen
Production:
[rH2] ≅ 0.0366 g/L-h
*Growth Variables Listed in
Literature Review Section*
XBi = 0.01 g/L
Si = 10 g/L
Figure 7. Predicted BIOMASS CSTR Outcome
30. Design Methodology: Evaluation of Alternatives
CSTR:
Hydrogen Production
132 kg per year
Operation Time
Continuously
120 days
Cleaning of reactor
Once a year
Loading procedures
Fill Storage
Minimal labor costs
Small volume
Small Storage
Batch:
Hydrogen Production
132 kg per year
Operation Time
10 hrs per day
120 days
Cleaning of reactor
Daily
Loading procedures
Daily
High labor costs
Large volume
Large storage
31. Design Methodology: Experimental Data
Experimental design
Final and initial temperature
measured using three different
flow rates
Heat transfer rate for each flow
rate calculated
Using a ratio, required solar panel
area calculated
Outside temperature 12.2℃
Cloudy; 12:30-2:30 P.M.
Flow Rate
[ml/min]
Inlet
Temp
[C]
Outlet
Temp [C]
Calculated q
[W]
569.62 18 19 40
172.74 18 30.2 130
79.86 18 43.1 120
Table 3. Solar Water Heater Experimental Data
32. Design Methodology: Synthesis of Design
Preheating Before Reaction:
Reactor should be close to 77° C
Solar heater used to preheat
Experimental values used to
achieve correct order of
magnitude
Solar water heater sized to be a
system of 5 panels at 4.5 m2
each
Sun's location
9:00 am ; 28.6° ; N 80.1° E
10:00 am ; 40.9° ; N 90.7° E Figure 2. Solar Water Heater Panel Dimensions
33. Design Methodology: Synthesis of Design
Heating throughout Reaction:
Reactor needs to be maintained at 77°C
Heating takes place for all 120 days
Loss of heat from outside
1.45 kW
4160.7 kW*hr
Night heating
10.9 kW
6,742.3 kW*hr
Water heating methods
Heating Jacket
Electric Heating https://sc01.alicdn.com/kf/HTB1xDKvGXXXXXXqXpXXq6xXF
XXX8/220798598/HTB1xDKvGXXXXXXqXpXXq6xXFXXX8.jp
g
37. Economic Analysis: CSTR Profits
Item Revenue (Total)
Hydrogen $300
Vinegar $6,250
Enzymes/Biomass Possible
Total $6,550
Table 7. CSTR Profits
38. Economic Analysis: Annual Worth Basis
Alternative 1: Do nothing
AWi=10%= -$43/ton * 66 tons/year = -$2840/year
Alternative 2: Fermentation Process
AWi=10%= -(A/P,i,10)($17175) - ($850+$3180)/year + $6550/year
AWi=10%= -($0.163*17175) + $2520/year
AWi=10%= -$280/year
Alternative Comparison
ΔAW2-1= -$280/year - (-$2840/year) = Savings of $2560/year
39. Sustainability Measures
Eliminate peach waste production by finding an alternate use for effluent flow
Reduce waste odors and reduce water pollution
Utilize waste peaches thrown into landfills
Use waste product to generate usable electricity
Reduce costs of energy production
Reduce use of fossil fuels and natural gases as sources for energy generation
40. Conclusion: Further Research
Acetic acid separation in downstream
Needed to make economically viable
Finding use for hyperthermophilic enzymes
Could generate large profits
Purification of gas effluent stream
Need for hydrogen fuel cell use
Actual implementation of design
Large troubleshooting necessary
Efficiency of implemented scale up
41. Conclusion: Questions Answered
User (Cherry Crossing or Musser Farm Operator)
How long does the operation take?
Continuous process for the 120 day growing season
What do I do with the reactor waste?
Salts: recycled
Water: recycled/compost
Acetic Acid: sold as vinegar
Biomass: extracted and sold
Peach Solids: added to compost
What components added to the reactor?
Peach slurry, water, salts, nitrogen source, pH controller, and Thermotoga neopalitana
42. Conclusion: Answering Questions
Client (Cherry Crossing or Musser Farm Manager)
How much will the implementation cost?
$17,175 implementation cost
How much money will it save us?
$2,560 annual savings if waste not sent to a landfill
How much energy can this process create on an annual basis?
The hydrogen gas will produce 2722 kW-hr per year
43. Conclusion: Answering Questions
Designer (Us)
How much waste is there needed to be processed?
60,000 kg (66.4 tons) of peach waste
How pure does the gas stream need to be?
Pure hydrogen is desirable for maximum fuel cell efficiency
How long does the reactor need to be in operation?
The reactor needs to be in operation for 120 days
44. Conclusion: Final Design
Final design selected was CSTR
120 days of operation
Hydrogen Production: 132 kg per year producing 2722 kW-hr per year
Effluents: Acetic acid, Water, Peach Solids, Salts, and Thermotoga neapolitana
Effluents Gas: Hydrogen, Nitrogen, Carbon Dioxide, Hydrogen Sulfide, and Water Vapor
Final Economics of CSTR
Initial Cost: $17,175
Material Cost: $850 annually
Running Cost: $3,180 annually
Profit: $6,550 annually
Total Annual Savings: $2,560
46. Acknowledgments
We would like to thank
Dr. Caye Drapcho,
Dr. Terry Walker,
Mr. Tom Jones,
and Mr. Jeff Hopkins
for their ongoing support and guidance throughout the project.
47. References / Patents
1. "1 Gallon Distilled White Vinegar - 4/Case." WebstaurantStore. N.p., 01 Sept. 2016. Web. 30 Nov. 2016.
2."1500l Semi-auto Ipa Beer Brewing Equipment,Cheap Price Beer Fermenting Equipment - Buy Beer Brewing Equipment,Ipa Beer Brewing Equipment,Semi-auto Ipa Beer Brewing Equipment Product on
Alibaba.com." Www.alibaba.com. N.p., n.d. Web. 30 Nov. 2016.
3."1500l Used Fermentation Tanks For Beer Fermenting And Maturing - Buy Used Fermentation Tanks,Used Fermentation Tanks,Used Fermentation Tanks Product on Alibaba.com." Www.alibaba.com. N.p., n.d.
Web. 30 Nov. 2016.
4."200l Stainless Steel Pot Cooking Equipment Jacketed Fermenter - Buy 200l Stainless Steel Pot,Cooking Equipment,Jacketed Fermenter Product on Alibaba.com." Www.alibaba.com. N.p., n.d. Web. 30 Nov.
2016.
5."AMT 368A-95 3/4" X 1/2" Cast Iron Straight Centrifugal Pump , Buna-N Seal, 1/3hp 1 Phase Motor." Global Industrial. N.p., n.d. Web. 30 Nov. 2016.
6."Current Canola Oil, Meal, and Seed Prices." Current Canola Oil, Meal, and Seed Prices - Canola Council of Canada. N.p., n.d. Web. 30 Nov. 2016.
7."Details: DSM-4359." Details: DSM-4359. N.p., n.d. Web. 30 Nov. 2016.
8."HSDB." U.S National Library of Medicine. U.S. National Library of Medicine, n.d. Web. 14 Oct. 2016.
9."Industrial Mixing Equipment - Product - 1 HP Electric Direct Drive Plate Mount Mixer by INDCO, New Albany, Indiana." Industrial Mixing Equipment - Product - 1 HP Electric Direct Drive Plate Mount Mixer by
INDCO, New Albany, Indiana. N.p., n.d. Web. 30 Nov. 2016.
10."Morton® Plain Table Salt - 25 Lb. Bag." Morton® Plain Table Salt - 25 Lb. Bag - Sam's Club. N.p., n.d. Web. 30 Nov. 2016.
11."Peach." Provital Group. Provital Group, 2008. Web.
12."Variety Display - Red Haven Peach Variety Evaluation." Musser Fruit Research Farm. Clemson University Horticulture, 2015. Web. 19 Oct. 2016.
13."You Must Pass a State Name!" USDA. USDA, 19 Oct. 2016. Web. 19 Oct. 2016.
14.Billing Administrations (n.d.): n. pag. City Of Clemson. City of Clemson, 1 July 2016. Web. 30 Nov. 2016.
15.Drapcho, C. M., Nhuan, N. P., & Walker, T. H. (2008). Biofuels engineering process technology. New York: McGraw Hill.
16.Hill, Louis, "Improvements in the process of biohydrogen production by Thermotoga neapolitana" (2013). All Theses. Paper 1796
17.Hills, D. J., & Roberts, D. W. (1982). Conversion of tomato, peach and honeydew solid waste into methane GAs. Transactions of the ASABE, 25(3), 821-826. Doi:10.103031
18.Joshi, H., Walker, T. H., & Nghiem, J. Optimization of ethanol production from peach waste. Unpublished manuscript.
19.Kukelko, D. A., D. S. Jayas, N.D. G. White, and M. G. Britton. "Physical Properties of Canola (rapeseed) Meal." Department of Agricultural Engineering (1987): n. pag. University of Manitoba. Web.
20.Newkirk, Rex, Ph.D. Canola Meal Feed Industry Guide (2009): 1-48. Print.
21.Walker, T. H. Utilization of peach's solid residue for enzyme production using trichoderma reesei rut C-30. Unpublished manuscript.
22.Wen, Ien-Chi, K. E. Koch, and W. B. Sherman. "Comparing Fruit and Tree Characteristics of Two Peaches and Their Nectarine Mutants." J. Amer. Soc. Hort. Sci. (1995): 101-06. Print.
23.Yu, X. 2007. Biohydrogen production by the hyperthermophilic bacterium Thermotoga neapolitana. PhD Dissertation, Clemson, South Carolina: Clemson University, Department of Environmental Engineering
and Earth Sciences
24.Yu, X., C.M. Drapcho. 2011. Hydrogen Production by the Hyperthermophilic Bacterium Thermotoga neapolitana using Agricultural-Based Carbon and Nitrogen Sources. Biological Engineering Transactions
2011;4:101-112.
65. Appendices: Study Period Determination
Table 16. Recovery Period of
Differing Asset Classes
Sullivan, William G., Elin M. Wicks, and C. Patrick Koelling.
Engineering Economy. Upper Saddle River, NJ: Pearson, 2015. Print.
