RRC ID 4918
Author Davis R, Anilkumar PK, Chandrashekar A, Shamala TR.
Title Biosynthesis of polyhydroxyalkanoates co-polymer in E. coli using genes from Pseudomonas and Bacillus.
Journal Antonie Van Leeuwenhoek
Abstract Expression of Pseudomonas aeruginosa genes PHA synthase1 (phaC1) and (R)-specific enoyl CoA hydratase1 (phaJ1) under a lacZ promoter was able to support production of a copolymer of Polyhydroxybutyrate (PHB) and medium chain length polyhydoxyalkanoates (mcl-PHA) in Escherichia coli. In order to improve the yield and quality of PHA, plasmid bearing the above genes was introduced into E. coli JC7623, harboring integrated beta-ketothiolase (phaA) and NADPH dependent-acetoacetyl CoA reductase (phaB) genes from a Bacillus sp. also driven by a lacZ promoter. The recombinant E. coli (JC7623ABC1J1) grown on various fatty acids along with glucose was found to produce 28-34% cellular dry weight of PHA. Gas chromatography and (1)H Nuclear Magnetic Resonance analysis of the polymer confirmed the ability of the strain to produce PHB-co-Hydroxy valerate (HV)-co-mcl-PHA copolymers. The ratio of short chain length (scl) to mcl-PHA varied from 78:22 to 18:82. Addition of acrylic acid, an inhibitor of beta-oxidation resulted in improved production (3-11% increase) of PHA copolymer. The combined use of enzymes from Bacillus sp. and Pseudomonas sp. for the production of scl-co-mcl PHA in E. coli is a novel approach and is being reported for the first time.
Volume 94(2)
Pages 207-16
Published 2008-8
DOI 10.1007/s10482-008-9233-3
PMID 18357511
MeSH Acetyl-CoA C-Acyltransferase / genetics Acetyl-CoA C-Acyltransferase / metabolism Acyltransferases / genetics Acyltransferases / metabolism Alcohol Oxidoreductases / genetics Alcohol Oxidoreductases / metabolism Bacillus / enzymology* Bacillus / genetics Bacterial Proteins / genetics Bacterial Proteins / metabolism Biomass Biopolymers / biosynthesis Biopolymers / chemistry Biopolymers / genetics Biosynthetic Pathways Escherichia coli / chemistry Escherichia coli / genetics Escherichia coli / growth & development Escherichia coli / metabolism* Gene Expression Genetic Engineering / methods* Polyhydroxyalkanoates / biosynthesis* Polyhydroxyalkanoates / chemistry Polyhydroxyalkanoates / genetics Pseudomonas / enzymology* Pseudomonas / genetics
IF 1.934
Times Cited 11
Prokaryotes E. coli pBRINT-Cm CV-38(JC7623)