FROM INDUSTRIAL WASTE GAS TO A GREEN ADDITIVE S y n g a s t o L i - - PowerPoint PPT Presentation
FROM INDUSTRIAL WASTE GAS TO A GREEN ADDITIVE S y n g a s t o L i q u i r i t i g e n i n NEXT SLIDE Meet Our Team Exotic Fermentation of Flavonoid Supervisor Team Captain Team member Enrico Orsi Xing Fu Adini Arifah Green Terpene:
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Exotic Fermentation of Flavonoid
Green Terpene: Sustainable production of terpenes by redesigning isoprene biosynthesis
Enrico Orsi
Supervisor Master student of Biotechnology Wageningen University
Xing Fu
Team Captain Master student of Biotechnology Wageningen University
Adini Arifah
Team member
Baking, brewing and cheese making in Egypt
First fermentation
Insulin for human produced by bacteria in America
First consumer product from GMO
The key entry enzyme that produce plant-based product expressed in yeast
First PAL gene expressed in microbes
Thermophilic strains Extreme PH/salty strains Syngas substrate strains We reaching a tipping point in traditional fermentation Biomass price increasing New energy Oil price dropping Contamination, less subsidy, more competitors...
Flavonoid Derivate
Green and non-toxic compound
Chemical industry
Natural food additives, cigarette additives
Food and consumer use
Anti-cancer1, anti-virus (HIV)1*
Pharmacy industry
From industrial waste gas to high value compound
Exotic pathway to fix CO2/H2 Exotic microbe to work in high temperature Gene modification to increase flux The pathway is working in E.coli and yeast
Pathway to Flavonoids (Liquiritigenin)
Ability to survive in 70 °C Able to fix CO2/H2/CO
Thermoanaerobacter kivui
From Acetyl-CoA to amino acid metabolism
From syngas to Acyl-CoA
Produce L-tyrosine
Final pathway to final product
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Investment6
Fermenter: 3M*2 € Gas cleaner: 0.2M*2 € Heat exchanger: 60k*2 €
DSP
Centrifuge:40k € Ultrafilter:50k € Distillation column: 50k*2 €
Cost of natural syngas
70 € /TCM7 (36041mol by 65 °C) 2k €/TCM7*Adding 10% H2
Production
17k € /TCM
Assumption C:H2 =3 5% carbon flux to product 416kg production 829g/mol
Cost and Return
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Based on 15 years
The current production of liquiritigenin is from plant. 660 € /kg (98%, China)
THE MARKET
Expected production 52t Net revenue 33 M € PMT:5%, 15 years 20 labours DSP excluded
THE ANNUAL REVENUE
10 70k ton market in China 2017 660 euro per Kg
Big market and high value
Possible to get syngas with low price/ subsidy Precursor for flavonoid production
Cheap substrate and promising DSP
By developing gene tools Higher yield is possible
Geno tools and yields
From Plant to bacteria
ATP related questions Flavonoid making cost ATP.
H2 is used to generate power to build Na+ gradient
Biegel, E., Schmidt, S., González, J. M., & Müller, V. (2011). Biochemistry, evolution and physiological function of the Rnf complex, a novel ion-motive electron transport complex in prokaryotes. Cellular and molecular life sciences, 68(4), 613-634.
Liquiritigenin is not soluble in cold water or methanol/ ethanol Crystal in acid liquid Crystal in pH2 Wash by methanol 98% Solution in MTBE (5v/v) Crystal by adding EDC 99.1%
Cost analysis
Gene enzyme
function effects Resource adhE alcohol/dehydrogenase delete Acyl-CoA competing pathway 2-fold of final product 8 TesB thioesterase II delete 10-fold of product 9 PDH* pyruvate dehydrogenase complex overexpress Malonyl-CoA pathway 60% increase of product 10 PGK phosphoglycerate kinase
GapA glyceraldehyde-3-phosphate dehydrogenase
FumC fumarase delete Malonyl-CoA competing pathway ACS Acyl-CoA synthase in WLP
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1. Ma, J., Fu, N. Y., Pang, D. B., Wu, W. Y., & Xu, A. L. (2001). Apoptosis induced by isoliquiritigenin in human gastric cancer MGC-803 cells. Planta medica, 67(08), 754-757. 1. *Harada, S. (2005). The broad anti-viral agent glycyrrhizin directly modulates the fluidity of plasma membrane and HIV-1
2. Metabolic engineering of microorganisms for the synthesis of plant natural products. Journal of biotechnology, 163(2), 166-178. 3. Flavonoid biosynthesis, wikipidia, https://en.wikipedia.org/wiki/Flavonoid_biosynthesis 4. Chen, G. Q., & Jiang, X. R. (2018). Next generation industrial biotechnology based on extremophilic bacteria. Current opinion in biotechnology, 50, 94-100. 5. Phillips, J. R., Huhnke, R. L., & Atiyeh, H. K. (2017). Syngas fermentation: a microbial conversion process of gaseous substrates to various
6. NEO programme of SenterNovem (October 2005), Bio-ethanol from bio-syngas , TU/e 7. Pei, P., Korom, S. F., Ling, K., & Nasah, J. (2016). Cost comparison of syngas production from natural gas conversion and underground coal
7.* INDEPENDENT REPORT TO THE DUTCH GOVERNMENT- Global sustainability objectives require a technological breakthrough, http://www.h2- fuel.nl/en/h2fuel_pdf/independent-report-dutch-government/ 8. Zhou, S., Iverson, A.G., Grayburn, W.S., 2008. Engineering a native homoethanol pathway in Escherichia coli B for ethanol production. Biotechnol.
9. Baek, J.M., Mazumdar, S., Lee, S.W., Jung, M.Y., Lim, J.H., Seo, S.W., Jung, G.Y., Oh, M.K., 2013. Butyrate production in engineered Escherichia coli with synthetic scaffolds. Biotechnol. Bioeng. 110, 2790–2794.
genetic inter- ventions predicted by OptForce computational framework. Chem. Eng. Sci. 103, 109–114. doi: 10.1016/j.ces.2012.10.009
acetobutylicum as Demonstrated by a Novel In Vivo CO Exchange Activity En Route to Heterologous Installation of a Functional Wood-Ljungdahl