Cloning and expression of the PHA synthase gene from a locally isolated Chromobacterium sp. USM2

Chromobacterium sp. USM2, a locally isolated bacterium was found to synthesize poly(3-hydroxybutyrate-co-3hydroxyvalerate), P(3HB-co-3HV) copolymer with high 3HV monomer composition. The PHA synthase gene was cloned and expressed in Cupriavidus necator PHB _ 4 to investigate the possibilities of incorporating other monomer. The recombinant successfully incorporated 3-hydroxyhexanoate (3HHx) monomer when fed with crude palm kernel oil (CPKO) as the sole carbon source. Approximately 63 +/- 2 wt% of P(3HB-co-3HHx) copolymer with 4 mol% of 3HHx was synthesized from 5 g/L of oil after 48 h of cultivation. In addition, P(3HB-co-3HV-co-3HHx) terpolymer with 9 mol% 3HV and 4 mol% 3HHx could be synthesized with a mixture of CPKO and sodium valerate. The presence of 3HV and 3HHx monomers in the copolymer and terpolymer was further confirmed with H+-NMR analysis. This locally isolated PHA synthase has demonstrated its ability to synthesize P(3HB-co-3HHx) copolymer from a readily available and renewable carbon source; CPKO, without the addition of 3HHx precursors.