1. Construction of Efficient Platform Escherichia coli Strains for Polyhydroxyalkanoate Production by Engineering Branched Pathway
- Author
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Byung-Gee Kim, Shashi Kant Bhatia, Hye-Rim Jung, Su-Yeon Yang, Ranjit Gurav, Eun Jung Kim, Hun-Suk Song, Yung-Hun Yang, Tae-Rim Choi, and Yu-Mi Moon
- Subjects
0106 biological sciences ,Polymers and Plastics ,Microorganism ,medicine.disease_cause ,01 natural sciences ,Polyhydroxyalkanoates ,Metabolic engineering ,lcsh:QD241-441 ,03 medical and health sciences ,lcsh:Organic chemistry ,010608 biotechnology ,medicine ,Escherichia coli ,NADPH ,CRISPR ,Gene ,CRISPR/Cas9 ,030304 developmental biology ,0303 health sciences ,Strain (chemistry) ,Chemistry ,Cas9 ,polyhydroxyalkanoate ,General Chemistry ,Biochemistry ,carbon flux distribution ,metabolic engineering - Abstract
Polyhydroxyalkanoate (PHA) is a potential substitute for petroleum-based plastics and can be produced by many microorganisms, including recombinant Escherichia coli. For efficient conversion of substrates and maximum PHA production, we performed multiple engineering of branched pathways in E. coli. We deleted four genes (pflb, ldhA, adhE, and fnr), which contributed to the formation of byproducts, using the CRISPR/Cas9 system and overexpressed pntAB, which catalyzes the interconversion of NADH and NADPH. The constructed strain, HR002, showed accumulation of acetyl-CoA and decreased levels of byproducts, resulting in dramatic increases in cell growth and PHA content. Thus, we demonstrated the effects of multiple engineering for redirecting carbon flux into PHA production without any concerns regarding simultaneous deletion.
- Published
- 2019
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