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Biosynthesis of polyhydroxyalkanoates co-polymer in E. coli using genes from Pseudomonas and Bacillus.
Reeta, Davis and Anil Kumar, P. K. and Chandrashekar, A. and Shamala, T. R. (2008) Biosynthesis of polyhydroxyalkanoates co-polymer in E. coli using genes from Pseudomonas and Bacillus. Antonie van Leeuwenhoek, 94 (2). pp. 207-16. ISSN 0003-6072
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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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Acetoacetyl CoA reductase � Acrylic acid � Bacillus sp. � b-Ketothiolase � E. coli � Enoyl CoA hydratase � PHA copolymers � PHA synthase � Pseudomonas sp. |
| Subjects: | 500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 27 Polymer Chemistry 500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology > 02 Bacteriology |
| Divisions: | Food Microbiology Plant Cell Biotechnology |
| Depositing User: | Food Sci. & Technol. Information Services |
| Date Deposited: | 16 Jun 2009 06:41 |
| Last Modified: | 22 May 2012 10:59 |
| URI: | http://ir.cftri.res.in/id/eprint/1802 |
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