Abstract
A group of bacilli phenotypically screened for synthesis and intracellular accumulation of PHAs granules by the use of Sudan Black B stain, eight strains were detected. Pair of specific PCR primers was designed and applied for genotypic detection of phaC synthase gene. Approximately, 760 bp DNA fragment was successfully amplified in the eight strains. Among the positive strains, Bacillus sp SW1-2, produced 36 g/L of the biopolymer during growth on modified E2 medium supplemented with glucose. Spectroscopic analysis by (CNMR)-N-13 and (HNMR)-N-1 revealed four narrow peaks (lines) (CH3; 21.2 ppm, CH2; 42.7, CH; 68.5 and C=O; 169.7 ppm) and 3 groups of signals (2.45, 2.58 and 5.2 ppm) identical and characteristic to polyhydroxybutyrate (PHB); respectively. Furthermore, the amplied PCR fragment, from genomic DNA of Bacillus sp SW1-2, was cloned in pGEM-T-Easy vector and sequenced with universal T7 and SP6 primers. The sequence showed 99% identity to phaC gene for polyhydroxyalkanoate synthase of many B. megaterium strains deposited in Genbank. While, showed 73% and 72% identity to synthases of Bacillus mycoides and Bacillus sp. INT005, respectively. [Mahmoud M. Berekaa. Genotyic Detection of Polyhydroxyalkanoate-producing Bacilli and Characterization of phaC Synthase of Bacillus sp. SW1-2. Life Sci J 2012;9(4):518-529] (ISSN:1097-8135). http://www.lifesciencesite.com. 78