Biosynthesis of polyhydroxyalkanoates co-polymer in E. coli using genes from Pseudomonas and Bacillus
- 21 March 2008
- journal article
- research article
- Published by Springer Science and Business Media LLC in Antonie van Leeuwenhoek
- Vol. 94 (2), 207-216
- https://doi.org/10.1007/s10482-008-9233-3
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 β-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 1H 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 β-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.Keywords
This publication has 39 references indexed in Scilit:
- Role of (R)-specific enoyl coenzyme A hydratases of Pseudomonas sp in the production of polyhydroxyalkanoatesAntonie van Leeuwenhoek, 2007
- The application of polyhydroxyalkanoates as tissue engineering materialsBiomaterials, 2005
- Molecular and Material Design of Biodegradable Poly(hydroxyalkanoate)sChemInform, 2004
- Engineering of Ralstonia eutropha for Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from Fructose and Solid-State Properties of the CopolymerBiomacromolecules, 2002
- Industrial scale production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)Applied Microbiology and Biotechnology, 2001
- A pBRINT family of plasmids for integration of cloned DNA into the Escherichia coli chromosomeGene, 1996
- Biosynthesis and characterization of hydroxybutyrate-hydroxycaproate copolymersInternational Journal of Biological Macromolecules, 1995
- Microbial Synthesis and Characterization of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)Macromolecules, 1995
- The role of fatty acid biosynthesis and degradation in the supply of substrates for poly(3-hydroxyalkanoate) formation inPseudomonas putidaFEMS Microbiology Letters, 1992
- Pseudomonas oleovorans as a Source of Poly(β-Hydroxyalkanoates) for Potential Applications as Biodegradable PolyestersApplied and Environmental Microbiology, 1988