Synergy of valine and threonine supplementation on poly(2-hydroxybutyrate-block-3-hydroxybutyrate) synthesis in engineered Escherichia coli expressing chimeric polyhydroxyalkanoate synthase

Sudo, Maho; Hori, Chiaki; Ooi, Toshihiko; Mizuno, Shoji; Tsuge, Takeharu; Matsumoto, Ken'ichiro

Synergy of valine and threonine supplementation on poly(2-hydroxybutyrate-block-3-hydroxybutyrate) synthesis in engineered Escherichia coli expressing chimeric polyhydroxyalkanoate synthase

Sudo, Maho; Hori, Chiaki; Ooi, Toshihiko et al.

Journal of Bioscience and Bioengineering, 2020, 129(3), 302-306

Permalink : https://hdl.handle.net/2115/80504

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アクセス権	open access
URI	https://hdl.handle.net/2115/80504
タイトル	Synergy of valine and threonine supplementation on poly(2-hydroxybutyrate-block-3-hydroxybutyrate) synthesis in engineered Escherichia coli expressing chimeric polyhydroxyalkanoate synthase
著者	Sudo, Maho ; Hori, Chiaki ; Ooi, Toshihiko ; Mizuno, Shoji ; Tsuge, Takeharu ; Matsumoto, Ken'ichiro
著者名 Sudo, Maho
e-Rad_Researcher 50722948 著者名 Hori, Chiaki
e-Rad_Researcher 40223713 著者名 Ooi, Toshihiko
著者名 Mizuno, Shoji
著者名 Tsuge, Takeharu
e-Rad_Researcher 80360642 著者名 Matsumoto, Ken'ichiro
言語	英語
キーワード	Block copolymer
	Biodegradable plastic
	Bio-based plastics
	Sequence regulation
	Monomer supply
発行日	2020-03
出版者	Elsevier
収録物名	Journal of Bioscience and Bioengineering
巻(号)	129(3)
ページ	302-306
抄録	The engineered chimeric polyhydroxyalkanoate (PHA) synthase PhaCAR is composed of N-terminal portion of Aeromonas caviae PHA synthase and C-terminal portion of Ralstonia eutropha (Cupriavidus necator) PHA synthase. PhaCAR has a unique and useful capacity to synthesize the block PHA copolymer poly(2-hydroxybutyrate-block-3-hydroxybutyrate) [P(2HB-b-3HB)] in engineered Escherichia coli from exogenous 2HB and 3HB. In the present study, we initially attempted to incorporate the amino acid-derived 2-hydroxyalkanoate (2HA) units using PhaCAR and the 2HA-CoA-supplying enzymes lactate dehydrogenase (LdhA) and CoA transferase (HadA). Cells harboring the genes for PhaCAR, LdhA, and HadA, as well as for the 3HB-CoA-supplying enzymes β-ketothiolase and acetoacetyl-CoA reductase, were cultivated with supplementation of four hydrophobic amino acids, i.e., leucine, valine (Val), isoleucine (Ile), and phenylalanine, in the medium. No hydrophobic amino acid-derived monomers were incorporated into the polymer, which was most likely because of the strict substrate specificity of PhaCAR; however, P(2HB-co-3HB) was unexpectedly produced with Val supplementation. The copolymer was likely P(2HB-b-3HB) based on proton nuclear magnetic resonance analysis. Based on the endogenous pathways in E. coli, 2HB units are likely derived from threonine (Thr) through deamination and dihydroxylation. In fact, dual supplementation with Thr and Val showed synergy on the 2HB fraction of the polymer. Val supplementation promoted the 2HB synthesis likely by inhibiting the metabolism of 2-ketobutyrate into Ile and/or activating Thr dehydratase. In conclusion, the LdhA/HadA/PhaCAR pathway served as the system for the synthesis of P(2HB-b-3HB) from biomass-derived carbon sources.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
権利情報	Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
権利情報	©2020. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
PISSN	1389-1723
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.jbiosc.2019.09.018