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Improved Butanol Production Using FASII Pathway in E. coli.
Jawed, Kamran; Abdelaal, Ali Samy; Koffas, Mattheos A G; Yazdani, Syed Shams.
Afiliação
  • Jawed K; Microbial Engineering Group, International Centre for Genetic Engineering and Biotechnology, 110067 New Delhi, India.
  • Abdelaal AS; Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, New York 12180, United States.
  • Koffas MAG; Microbial Engineering Group, International Centre for Genetic Engineering and Biotechnology, 110067 New Delhi, India.
  • Yazdani SS; Department of Genetics, Faculty of Agriculture, Damietta University, 34511 Damietta, Egypt.
ACS Synth Biol ; 9(9): 2390-2398, 2020 09 18.
Article em En | MEDLINE | ID: mdl-32813973
n-Butanol is often considered a potential substitute for gasoline due to its physicochemical properties being closely related to those of gasoline. In this study, we extend our earlier work to convert endogenously producing butyrate via the FASII pathway using thioesterase TesBT to its corresponding alcohol, i.e., butanol. We first assembled pathway genes, i.e., car encoding carboxylic acid reductase from Mycobacterium marinum, sfp encoding phosphopantetheinyl transferase from Bacillus subtilis, and adh2 encoding alcohol dehydrogenase from S. cerevisiae, responsible for bioconversion of butyrate to butanol in three different configurations (Operon, Pseudo-Operon, and Monocistronic) to achieve optimum expression of each gene and compared with the clostridial solventogenic pathway for in vivo conversion of butyrate to butanol under aerobic conditions. An E. coli strain harboring car, sfp, and adh2 in pseudo-operon configuration was able to convert butyrate to butanol with 100% bioconversion efficiency when supplemented with 1 g/L of butyrate. Further, co-cultivation of an upstream strain (butyrate-producing) with a downstream strain (butyrate to butanol converting) at different inoculation ratios was investigated, and an optimized ratio of 1:4 (upstream strain: downstream strain) was found to produce ∼2 g/L butanol under fed-batch fermentation. Further, a mono-cultivation approach was applied by transforming a plasmid harboring tesBT gene into the downstream strain. This approach produced 0.42 g/L in a test tube and ∼2.9 g/L butanol under fed-batch fermentation. This is the first report where both mono- and co-cultivation approaches were tested and compared for butanol production, and butanol titers achieved using both strategies are the highest reported values in recombinant E. coli utilizing FASII pathway.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: 1-Butanol / Escherichia coli / Vias Biossintéticas / Engenharia Metabólica Idioma: En Revista: ACS Synth Biol Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Índia

Texto completo: 1 Base de dados: MEDLINE Assunto principal: 1-Butanol / Escherichia coli / Vias Biossintéticas / Engenharia Metabólica Idioma: En Revista: ACS Synth Biol Ano de publicação: 2020 Tipo de documento: Article País de afiliação: Índia