Real-time NMR analysis of polyhydroxyalkanoate synthase reaction that synthesizes block copolymer comprising glycolate and 3-hydroxybutyrate

Yanagawa, Kengo; Kajikawa, Ayaka; Sakakibara, Sayaka; Kumeta, Hiroyuki; Tomita, Hiroya; Matsumoto, Ken'ichiro

Real-time NMR analysis of polyhydroxyalkanoate synthase reaction that synthesizes block copolymer comprising glycolate and 3-hydroxybutyrate

Yanagawa, Kengo; Kajikawa, Ayaka; Sakakibara, Sayaka et al.

Biophysical Chemistry, 2023, 296, 107001

Permalink : https://hdl.handle.net/2115/94145

このアイテムのアクセス数:175件（2026-08-17 23:46 集計）

閲覧可能ファイル

ファイル	フォーマット	サイズ	閲覧回数	説明
RevisedManuscript0328	pdf	2.39 MB	145

論文情報

ファイル出力 EndNote Basic出力 Mendeley出力

アクセス権	open access
URI	https://hdl.handle.net/2115/94145
タイトル	Real-time NMR analysis of polyhydroxyalkanoate synthase reaction that synthesizes block copolymer comprising glycolate and 3-hydroxybutyrate
著者	Yanagawa, Kengo ; Kajikawa, Ayaka ; Sakakibara, Sayaka ; Kumeta, Hiroyuki ; Tomita, Hiroya ; Matsumoto, Ken'ichiro
著者名 Yanagawa, Kengo
著者名 Kajikawa, Ayaka
著者名 Sakakibara, Sayaka
著者名 Kumeta, Hiroyuki
e-Rad_Researcher 00814229 著者名 Tomita, Hiroya
e-Rad_Researcher 80360642 著者名 Matsumoto, Ken'ichiro
キーワード	Polyhydroxybutyrate
	Enzymatic polymerization
	Block copolymer
	Glycolate-based polymer
	Polyglycolic acid
発行日	2023-05
出版者	Elsevier
収録物名	Biophysical Chemistry
巻(号)	296
ページ	107001
抄録	The sequence-regulating polyhydroxyalkanoate (PHA) synthase PhaCAR spontaneously synthesizes the homo-random block copolymer, poly[3-hydroxybutyrate (3HB)]-b-poly[glycolate (GL)-ran-3HB]. In this study, a real-time in vitro chasing system was established using a high-resolution 800 MHz nuclear magnetic resonance (NMR) and 13C-labeled monomers to monitor the polymerization of GL-CoA and 3HB-CoA into this atypical copolymer. Consequently, PhaCAR initially consumed only 3HB-CoA and subsequently consumed both substrates. The structure of the nascent polymer was analyzed by extracting it with deuterated hexafluoro-isopropanol. In the primary reaction product, a 3HB-3HB dyad was detected, and GL-3HB linkages were subsequently formed. According to these results, the P(3HB) homopolymer segment is synthesized prior to the random copolymer segment. This is the first report of its kind which proposes the application of real-time NMR to a PHA synthase assay, paving the way for elucidating the mechanisms of PHA block copolymerization.
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	© <2023>. 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	0301-4622
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.bpc.2023.107001