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In silico design and adaptive evolution of Escherichia coli for production of lactic acid

In silico design and adaptive evolution of Escherichia coli for production of lactic acid. Presented by: Samantha Gonyea S.S. Fong, A.P. Burgard, C.D. Herring, E.M. Knight, F.R. Blattner, C.D. Maranas, & B.O. Palsson 2005. Biotechnology and Bioengineering 91: 643-648. C. E. A. B. D.

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In silico design and adaptive evolution of Escherichia coli for production of lactic acid

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  1. In silico design and adaptive evolution of Escherichia coli for production of lactic acid Presented by: Samantha Gonyea S.S. Fong, A.P. Burgard, C.D. Herring, E.M. Knight, F.R. Blattner, C.D. Maranas, & B.O. Palsson 2005. Biotechnology and Bioengineering 91: 643-648.

  2. C E A B D What is metabolic engineering? • The manipulation of an organism for a specific purpose such as increases in certain metabolites • Often accomplished by deleting genes of interest to “force” the usage of a different metabolic pathway X X

  3. What was the project goal? • To develop an E. coli strain to produce lactate • How was this accomplished? • Using computer models • Experimentally testing strains www.igmors.u-psud.fr

  4. What were some hypotheses? • Computer models would help to determine the strain designs that would optimize growth and lactate production • E. coli would undergo adaptive evolution by finding alternate metabolic pathways • Lactate production rate would increase as growth rate increased

  5. What did the computer models tell us? • OptKnock algorithm solutions • Optimization of biomass formation and lactate secretion • iJE660 E. coli metabolism model used for three strain designs • All strains began with wild-type E. coli strain K12 MG1655 www.rowett.ac.uk

  6. Glucose glk F6P pfk PEP TCA Cycle Lactate Pyruvate ACCoA adhE pta Ethanol Acetate Metabolic Pathway of Lactate • pta-adhE strain X X

  7. Glucose glk F6P pfk PEP TCA Cycle Lactate Pyruvate ACCoA adhE pta Ethanol Acetate Metabolic Pathway of Lactate • pta-pfk strain X X

  8. Glucose glk F6P pfk PEP TCA Cycle Lactate Pyruvate ACCoA adhE pta Ethanol Acetate Metabolic Pathway of Lactate X • pta-adhE-pfk-glk strain X X X

  9. Results of Adaptive Evolution

  10. Results of Adaptive Evolution • The computer model predictions were successfully tested in vitro • Adaptive evolution can lead to an increase in a particular metabolite • Stable strains for production of metabolites can be achieved using these techniques www.rib.okayama-u.ac.jp

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