Combination of N149S and D171G mutations in Aeromonas caviae polyhydroxyalkanoate synthase and impact on polyhydroxyalkanoate biosynthesis.

Combination of N149S and D171G mutations in Aeromonas caviae polyhydroxyalkanoate synthase and impact on polyhydroxyalkanoate biosynthesis. Research Abstract Details 

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Combination of N149S and D171G mutations in Aeromonas caviae polyhydroxyalkanoate synthase and impact on polyhydroxyalkanoate biosynthesis. Abstract Text:

Takeharu Tsuge,Shinko Watanabe,Daisuke Shimada,Hideki Abe,Yoshiharu Doi,Seiichi Taguchi,Takeharu Tsuge,Shinko Watanabe,Daisuke Shimada,Hideki Abe,Yoshiharu Doi,Seiichi Taguchi,Takeharu Tsuge,Shinko Watanabe,Daisuke Shimada,Hideki Abe,Yoshiharu Doi,Seiichi Taguchi,

Aeromonas caviae polyhydroxyalkanoate synthase (PhaC(Ac)) is an important biocatalyst for the synthesis of practically useful two-component polyhydroxyalkanoate copolymer, poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] [P(3HB-co-3HHx)]. In a previous study, two PhaC(Ac) mutants that have a single amino acid substitution of either asparagine 149 by serine (N149S) or aspartate 171 by glycine (D171G) were isolated as higher active enzymes by means of evolutionary engineering. In this study, the synergistic effects of N149S and D171G double mutation (NSDG) in PhaC(Ac) on polyhydroxyalkanoate biosynthesis were investigated in recombinant Ralstonia eutropha. The PhaC(Ac) NSDG mutant showed enhanced incorporation of longer 3-hydroxyalkanoate (3HA) units into the polyhydroxyalkanoate copolymer from octanoate (3HA fraction: 18.5 mol%) and soybean oil (5.4 mol%) as a carbon source. Besides, the NSDG mutant synthesized P(3HB) homopolymer with a very high molecular weight (M(w)=368 x 10(4)) when fructose was used as a carbon source. Thus, a combination of the beneficial mutations synergistically altered enzymatic properties, leading to synthesis of a polyhydroxyalkanoate copolymer with enhanced 3HA fraction and increased molecular weight.

Combination of N149S and D171G mutations in Aeromonas caviae polyhydroxyalkanoate synthase and impact on polyhydroxyalkanoate biosynthesis. Publishing Authors By Initials

T Tsuge,S Watanabe,D Shimada,H Abe,Y Doi,S Taguchi,T Tsuge,S Watanabe,D Shimada,H Abe,Y Doi,S Taguchi,T Tsuge,S Watanabe,D Shimada,H Abe,Y Doi,S Taguchi,

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Combination of N149S and D171G mutations in Aeromonas caviae polyhydroxyalkanoate synthase and impact on polyhydroxyalkanoate biosynthesis. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: FEMS microbiology letters

VOLUME: 277

Page Numbers: 217-22

Journal Abbreviation: FEMS Microbiol. Lett.

ISSN: 0378-1097

DAY: 22

MONTH: Dec

YEAR: 2007

Combination of N149S and D171G mutations in Aeromonas caviae polyhydroxyalkanoate synthase and impact on polyhydroxyalkanoate biosynthesis. Information

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LANGUAGE: eng

NlmUniqueID: 7705721

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Grant and Affiliation Information for Combination of N149S and D171G mutations in Aeromonas caviae polyhydroxyalkanoate synthase and impact on polyhydroxyalkanoate biosynthesis.

AFFILIATION: Department of Innovative and Engineered Materials, Tokyo Institute of Technology, Yokohama, Japan.

Country: England

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MEDLINETA: FEMS Microbiol Lett

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