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Improved production of poly(lactic acid)-like polyester based on metabolite analysis to address the rate-limiting step

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Title: Improved production of poly(lactic acid)-like polyester based on metabolite analysis to address the rate-limiting step
Authors: Matsumoto, Ken'ichiro Browse this author →KAKEN DB
Tobitani, Kota Browse this author
Aoki, Shunsuke Browse this author
Song, Yuyang Browse this author
Ooi, Toshihiko Browse this author →KAKEN DB
Taguchi, Seiichi Browse this author →KAKEN DB
Keywords: Biobased plastic
Metabolome analysis
Issue Date: 18-Nov-2014
Publisher: BioMed Central
Journal Title: AMB express
Volume: 4
Start Page: 83
Publisher DOI: 10.1186/s13568-014-0083-2
Abstract: The biosynthesis of poly(lactic acid) (PLA)-like polymers, composed of >99 mol% lactate and a trace amount of 3-hydroxybutyrate, in engineered Corynebacterium glutamicum consists of two steps; the generation of the monomer substrate lactyl-coenzyme A (CoA) and the polyhydroxyalkanoate (PHA) synthase-catalyzed polymerization of lactyl-CoA. In order to increase polymer productivity, we explored the rate-limiting step in PLA-like polymer synthesis based on quantitative metabolite analysis using liquid chromatography mass spectroscopy (LC-MS). A significant pool of lactyl-CoA was found during polymer synthesis. This result suggested that the rate-limitation occurred at the polymerization step. Accordingly, the expression level of PHA synthase was increased by means of codon-optimization of the corresponding gene that consequently led to an increase in polymer content by 4.4-fold compared to the control. Notably, the codon-optimization did not significantly affect the concentration of lactyl-CoA, suggesting that the polymerization reaction was still the rate-limiting step upon the overexpression of PHA synthase. Another important finding was that the generation of lactyl-CoA was concomitant with a decrease in the acetyl-CoA level, indicating that acetyl-CoA served as a CoA donor for lactyl-CoA synthesis. These results show that obtaining information on the metabolite concentrations is highly useful for improving PLA-like polymer production. This strategy should be applicable to a wide range of PHA-producing systems.
Type: article
Appears in Collections:工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 田口 精一

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