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Production of Malonic Acid Through the Fermentation of Glucose

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Production of Malonic Acid Through the Fermentation of Glucose University of Pennsylvania ScholarlyCommons Department of Chemical & Biomolecular Senior Design Reports (CBE) Engineering 4-20-2018 Production of Malonic Acid through the Fermentation of Glucose Emily P. Peters University of Pennsylvania, [email protected] Gabrielle J. Schlakman University of Pennsylvania, [email protected] Elise N. Yang University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/cbe_sdr Part of the Biochemical and Biomolecular Engineering Commons Peters, Emily P.; Schlakman, Gabrielle J.; and Yang, Elise N., "Production of Malonic Acid through the Fermentation of Glucose" (2018). Senior Design Reports (CBE). 107. https://repository.upenn.edu/cbe_sdr/107 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/cbe_sdr/107 For more information, please contact [email protected]. Production of Malonic Acid through the Fermentation of Glucose Abstract The overall process to produce malonic acid has not drastically changed in the past 50 years. The current process is damaging to the environment and costly, requiring high market prices. Lygos, Inc., a lab in Berkeley, California, has published a patent describing a way to produce malonic acid through the biological fermentation of genetically modified easty cells. This proposed technology is appealing as it is both better for the environment and economically friendly. For the process discussed in this report, genetically modified Pichia Kudriavzevii yeast cells will be purchased from the Lygos lab along with the negotiation of exclusive licensing rights to the technology. The cells will be grown in fermentation vessels, while being constantly fed oxygen, glucose and fermentation media. The cells will excrete malonic acid in the 101 hour fermentation process. In order to meet a production capacity of 10M pounds of malonic acid a year, 236 total batches are needed. The fermentation broth will then be fed continuously to a downstream process which includes vacuum filtration, reverse osmosis, and crystallization to produce a solid malonic acid powder. After drying, malonic acid crystal powder of 99.9% purity will be sold to the cosmetic, pharmaceutical and petrochemical industries. The design requires an initial investment of $23.1M. The investors’ rate of return (IRR) is 52.6%, the return on investment (ROI) is 46.9% in year three, and the net present value (NPV) is $59.6M in 2018. Sensitivity analyses on the licensing fee and price of cells concluded that these prices are negotiable with Lygos, Inc. This design is recommended based on the process specifications and economic viability of the process, but the success of this project largely depends on the agreement that can be reached with the originators of the technology, Lygos. Disciplines Biochemical and Biomolecular Engineering | Chemical Engineering | Engineering This working paper is available at ScholarlyCommons: https://repository.upenn.edu/cbe_sdr/107 Production of Malonic Acid Peters | Schlakman | Yang Dear Dr. John Vohs and Professor Bruce Vrana, The enclosed reports contains a proposal for the design of a process to produce 10 Million pounds per year of Malonic Acid from the biological fermentation of glucose using genetically engineered Pichia Kudriavzevii yeast cells. Our process will depend largely on a partnership with Lygos, Inc., a biotechnology company that has developed the technology to genetically modify yeast cells to produce large amounts of Malonic Acid. Vials of cells will be bought from this company and put into a seed fermentation line consisting of three seed fermenters for 16 hours each and finally into a production fermenter for 50 hours. To meet our production goal of 10 Million pounds per year, we will require three sets of seed lines and production fermenters. 236 batches will be produced in 47 operating weeks, or 329 days, equaling about 5 batches per week. Each batch will then be sent to a downstream process consisting of vacuum belt filtration, reverse osmosis, and crystallization to result in a final solid Malonic Acid product of 99.9% purity. To determine the economic viability of this proposed process, our team conducted a thorough economic analysis. We decided pursuing exclusive licensing rights with Lygos, Inc. would be the best route to pursue due to Lygos’ existing expertise in the technology and our desire to keep costs as low as possible. The economic feasibility of our process depended largely on the potential purchase price and licensing fee we would be able to negotiate with Lygos, and our base case is based on a purchase price of $5,000/vial of cells with an annual licensing fee of $1,000,000. With these values and a selling price of $5.00 per pound of Malonic Acid, as stated in our problem statement, our process will require a $23.1 Million total investment, providing an IRR of 52.6% and a third-year ROI of 46.9%. Sensitivity analyses on these costs and on selling price was performed, providing additional support to our conclusion of economic feasibility. The current price of Malonic Acid on the market ranges from $10-$100 per pound, giving us a significant competitive advantage if an agreement with Lygos can be reached. Thus, our team recommends investment in this process design. Sincerely, _________________ _________________ _________________ Emily Peters Gabrielle Schlakman Elise Yang 1 Production of Malonic Acid Peters | Schlakman | Yang 2 Production of Malonic Acid Peters | Schlakman | Yang Production of Malonic Acid through the Fermentation of Glucose Emily Peters | Gabrielle Schlakman | Elise Yang Project Submitted to Mr. Bruce Vrana and Dr. John Vohs Project Proposed by Mr. Bruce Vrana Department of Chemical and Biomolecular Engineering School of Engineering and Applied Science University of Pennsylvania April 17, 2018 3 Production of Malonic Acid Peters | Schlakman | Yang Table of Contents SECTION 1: ABSTRACT……………………………………………………………………....8 SECTION 2: INTRODUCTION………………..………………………………………………9 2.1 Background Information………………………………………………………….…...9 2.2 Objective Time Chart………………………………………………….……………..11 SECTION 3: INNOVATION MAP………………………...………………………………….13 SECTION 4: MARKET AND COMPETITIVE ANALYSIS.………………………………14 SECTION 5: CUSTOMER REQUIREMENTS.……………………………………………..17 SECTION 6: CRITICAL TO QUALITY VARIABLES.……………………………………18 SECTION 7: PRODUCT CONCEPT.……………………………………………………...…19 SECTION 8: SUPERIOR PRODUCT CONCEPT.………………………………………….20 SECTION 9: COMPETITIVE ANALYSIS.………………………………………………….20 SECTION 10: PRELIMINARY PROCESS SYNTHESIS..…………………………………21 10.1 Yeast Cells.…………………………………………………………………………21 10.2 Production Fermenter Sizing……………………………………………………….24 10.3 Batch vs Continuous Downstream Processing……………………………………..24 10.4 Removal of Biomass from Fermentation Broth………………………………….....25 10.5 Filtration Rinse……………………………………………………………………..26 10.6 Increasing the Malonic Acid Concentration before Crystallization………………..26 10.7 Crystallization of Malonic Acid.…………………………………………………....27 10.8 Drying of Final Product……………………………………………………...……..28 10.9 Downstream Vessel Sizing……………………………………………………..…..28 10.10 Downstream Pumps…………………………………………………………….....29 SECTION 11: ASSEMBLY OF A DATABASE………………………………………...……30 11.1 Material and Energy Components………………………………………….......…..30 11.2 Economic Components………………………………………………………….… 31 SECTION 12: PROCESS DESCRIPTION, PROCESS FLOW DIAGRAM, MATERIAL BALANCE…………………………………………………………...…………………………32 12.1 Overall Process……………………………………………………………………..32 12.2 Fermentation………………………………………………………………………..35 12.2.1 Lab Fermentation…………………………………………….…….……..35 12.2.2 Fermentation Reactions…………………………………………………..35 12.2.3 Seed fermenter 1………………………………………………………….38 12.2.4 Seed Fermenter 2………………………………………………..………..40 12.2.5 Seed Fermenter 3……………………………………………………..…..42 4 Production of Malonic Acid Peters | Schlakman | Yang 12.2.6 Production Fermenter……………………………………………………..45 12.2.7 Scheduling………………………………………………………….……..47 12.3 Separation and Purification.………………………………………………….……..50 12.3.1 Vacuum Belt Filter………………………………………………………..50 12.3.2 Reverse Osmosis…………………………………………………...……..51 12.3.3 Crystallizer………………………………………………………………..52 12.3.4 Vacuum Belt Filter………………………………………………………..53 12.3.5 Fluidized Dryer…………………………………………………………...54 12.3.6 Scheduling……………………………………………………………..….55 SECTION 13: ENERGY BALANCE AND UTILITY REQUIREMENTS……….………..56 13.1 Energy Balance……………………………………………………………………..56 13.1.1 Mixing Tanks……………………………………………………………..56 13.1.2 Heat of Reactions in Fermenters……………………...…………………..57 13.1.3 Fermenter Agitation………………………………………………..……..58 13.1.4 Pumps between Fermenters……………………………………………....59 13.1.5 Water Filter…………………………………………………………...…..60 13.1.6 Air Filter.………….………………………………………………..……..60 13.1.7 Scrubber…………………………………………………………………..61 13.1.8 Downstream Pumps……………………………………………..………..61 13.1.9 Vacuum Belt Filters………………………………………..……………..62 13.1.10 Dryer Air…………………………………………………………….…..63 13.2 Utility Requirements………………………………………………………………..63 13.2.1 Sterilization of YPD Media Through the Jacket………………..………..64 13.2.2 Sterilization of YNB Media Through Heat Exchanger Network…..……..65 13.2.3 Cooling the Fermentation Vessels………………………………………..67 13.2.4 Water as a Raw Material……………………………………………...…..67 13.2.5 Cooling of the Crystallizer………………………………………………..68 SECTION 14: EQUIPMENT LISTS, UNIT DESCRIPTIONS AND SPECIFICATION....69 14.1 Fermenters…………………………………………………………………………..69 14.2 Heat Exchangers……………………………………………………………..……..70 14.3 Pumps…………………………………………………………………………...…..71 14.4 Mixing Vessels…………………………………………………………….………..71
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