Evaluating the Ability of Polyhydroxyalkanoate Synthase Mutants to Produce P(3HB-co-3HA) from Soybean Oil

Polyhydroxyalkanoate (PHA) synthase from Pseudomonas sp 61-3 (PhaCl(Ps)) is able to synthesize P(3HB-co-MA), consisting of a 3HB unit and medium-chain-length 3HA units of 6-12 carbon atoms. Expression vectors encoding 76 PhaC1(Ps) mutants with an amino acid replacement at position 130, 325, 477 or 481 were individually introduced into Ralstonia eutropha. The mutant enzyme genes were evaluated in terms of their abilities to synthesize P(3HB-co-3HA) using soybean oil as a carbon source. 20 mutants showed significantly high accumulation levels of PHA exceeding 30 wt.-% and as high as 57 wt.-%. It was found that hydrophobic amino acids at the positions are more likely to enhance accumulation of PHA in R. eutropha.