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• W2100448885 endingPage "2419" @default.
• W2100448885 startingPage "2411" @default.
• W2100448885 abstract "ABSTRACT By in vitro evolution experiment, we have first succeeded in acquiring higher active mutants of a synthase that is a key enzyme essential for bacterial synthesis of biodegradable polyester, polyhydroxyalkanoate (PHA). Aeromonas caviae FA440 synthase, termed PhaC Ac , was chosen as a good target for evolution, since it can synthesize a PHA random copolyester of 3-hydroxybutyrate and 3-hydroxyhexanoate [P(3HB- co -3HHx)] that is a tough and flexible material compared to polyhydroxybutyrate (PHB) homopolyester. The in vitro enzyme evolution system consists of PCR-mediated random mutagenesis targeted to a limited region of the phaC Ac gene and screening mutant enzymes with higher activities based on two types of polyester accumulation system by using Escherichia coli for the synthesis of PHB (by JM109 strain) (S. Taguchi, A. Maehara, K. Takase, M. Nakahara, H. Nakamura, and Y. Doi, FEMS Microbiol. Lett. 198:65-71, 2001) and of P(3HB- co -3HHx) {by LS5218 [ fadR601 atoC (Con)] strain}. The expression vector for the phaC Ac gene, together with monomer-supplying enzyme genes, was designed to synthesize PHB homopolyester from glucose and P(3HB- co -3HHx) copolyester from dodecanoate. Two evolved mutant enzymes, termed E2-50 and T3-11, screened through the evolution system exhibited 56 and 21% increases in activity toward 3HB-coenzyme A, respectively, and consequently led to enhanced accumulation (up to 6.5-fold content) of P(3HB- co -3HHx) in the recombinant LS5218 strains. Two single mutations in the mutants, N149S for E2-50 and D171G for T3-11, occurred at positions that are not highly conserved among the PHA synthase family. It should be noted that increases in the 3HHx fraction (up to 16 to 18 mol%) were observed for both mutants compared to the wild type (10 mol%)." @default.
• W2100448885 created "2016-06-24" @default.
• W2100448885 creator A5027215968 @default.
• W2100448885 creator A5027898163 @default.
• W2100448885 creator A5032760176 @default.
• W2100448885 date "2002-05-01" @default.
• W2100448885 modified "2023-10-14" @default.
• W2100448885 title "Enhanced Accumulation and Changed Monomer Composition in Polyhydroxyalkanoate (PHA) Copolyester by In Vitro Evolution of <i>Aeromonas caviae</i> PHA Synthase" @default.
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• W2100448885 doi "https://doi.org/10.1128/aem.68.5.2411-2419.2002" @default.
• W2100448885 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/127549" @default.
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