Bio-Plastics PHA Production by Recombinant Bacillus Subtilis

Y.J. Wang, F.L. Hua, Y.F. Tsang, S.Y. Chan, H. Chua, and P.H.F. Yu (PRC)


Bacillus subtilis, cloning and expression, PHA synthase gene, P (HB-co-mclHA), Polyhydroxyalkanoates (PHAs)


Polyhydroxyalkanoates (PHAs) are polyesters of hydroxyalkanoates synthesized by numerous bacteria as intracellular carbon and energy storage compounds and accumulated as granules in the cytoplasm of cells. In this work, we constructed two recombinant plasmids, pBE2C1 and pBE2C1AB. The two plasmids were inserted into Bacillus subtilis DB104 and generated Bacillus subtilis/ pBE2C1 and Bacillus subtilis/pBE2C1AB. The two recombinant strains were subjected to fermentation and showed PHA accumulation, the first reported example of mcl-PHA production in Bacillus subtilis. GC analysis identified the compound produced by Bacillus subtilis/ pBE2C1 was a hydroxydecanoate -co hydroxydodecanoate (HD-co-HDD) polymer while that produced by Bacillus subtilis/pBE2C1AB was a hydroxybutyrate-co-hydroxydecanoate-co-hydroxydodeca noate (HB-HD-HDD) polymer. The results also showed that the recombinant B. subtilis could utilize the malt waste in the medium as a carbon source better than that of glucose and thus could substantially lowered the cost of production of PHA.

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