Real-time NMR analysis of polyhydroxyalkanoate synthase reaction that synthesizes block copolymer comprising glycolate and 3-hydroxybutyrate.
Real-time NMR analysis of polyhydroxyalkanoate synthase reaction that synthesizes block copolymer comprising glycolate and 3-hydroxybutyrate.
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合成包含乙醇酸酯和 3-羟基丁酸酯的嵌段共聚物的聚羟基链烷酸酯合酶反应的实时 NMR 分析。
DOI：
10.2139/ssrn.4325586
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发表时间：
2023
影响因子：
3.8
通讯作者：
Ken’ichiro Matsumoto
中科院分区：
文献类型：
--
作者：
Kengo Yanagawa;Ayaka Kajikawa;Sayaka Sakakibara;H. Kumeta;H. Tomita;Ken’ichiro Matsumoto
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.
影响因子：
6.4
作者：
通讯作者：
--
影响因子：
6.2
作者：
Matsusaki, H;Abe, H;Doi, Y
通讯作者：
Doi, Y
影响因子：
2.9
作者：
Jia, Y;Yuan, W;Stubbe, J
通讯作者：
Stubbe, J