RRC ID 35621
著者 Hiroe A, Hyakutake M, Thomson NM, Sivaniah E, Tsuge T.
タイトル Endogenous ethanol affects biopolyester molecular weight in recombinant Escherichia coli.
ジャーナル ACS Chem Biol
Abstract In biopolyester synthesis, polyhydroxyalkanoate (PHA) synthase (PhaC) catalyzes the polymerization of PHA in bacterial cells, followed by a chain transfer (CT) reaction in which the PHA polymer chain is transferred from PhaC to a CT agent. Accordingly, the frequency of CT reaction determines PHA molecular weight. Previous studies have shown that exogenous alcohols are effective CT agents. This study aimed to clarify the effect of endogenous ethanol as a CT agent for poly[(R)-3-hydroxybutyrate] [P(3HB)] synthesis in recombinant Escherichia coli, by comparing with that of exogenous ethanol. Ethanol supplementation to the culture medium reduced P(3HB) molecular weights by up to 56% due to ethanol-induced CT reaction. NMR analysis of P(3HB) polymers purified from the culture supplemented with (13)C-labeled ethanol showed the formation of a covalent bond between ethanol and P(3HB) chain at the carboxyl end. Cultivation without ethanol supplementation resulted in the reduction of P(3HB) molecular weight with increasing host-produced ethanol depending on culture aeration. On the other hand, production in recombinant BW25113(ΔadhE), an alcohol dehydrogenase deletion strain, resulted in a 77% increase in molecular weight. Analysis of five E. coli strains revealed that the estimated number of CT reactions was correlated with ethanol production. These results demonstrate that host-produced ethanol acts as an equally effective CT agent as exogenous ethanol, and the control of ethanol production is important to regulate the PHA molecular weight.
巻・号 8(11)
ページ 2568-76
公開日 2013-11-15
DOI 10.1021/cb400465p
PMID 24041146
MeSH Acyltransferases / metabolism Animals Cattle Cell Line, Tumor Cell Proliferation / drug effects Chromatography, Liquid Drug Evaluation, Preclinical Escherichia coli / chemistry Escherichia coli / enzymology Escherichia coli / genetics Escherichia coli / metabolism* Ethanol / metabolism* Ethanol / pharmacology* Humans Hydroxybutyrates / chemistry* Inhibitory Concentration 50 Molecular Structure Molecular Weight Polyesters / chemistry* Recombinant Proteins / genetics Recombinant Proteins / metabolism Structure-Activity Relationship
IF 4.434
引用数 18
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
原核生物(大腸菌) NA