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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 | | P(LA-co-3HB) | | Polyhydroxybutyrate | | 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. |
| Rights: | http://creativecommons.org/licenses/by/4.0 |
| Type: | article |
| URI: | http://hdl.handle.net/2115/59780 |
| 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)
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Submitter: 田口 精一
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