Biosynthesis and Characterization of Poly3-hydroxyalkanote (PHA) from Newly Isolated Bacterium Bacillus sp. AZR-1

Bacterial poly3-hydroxyalkanotes are the polyesters that are accumulated in conditions of un-balanced nutrients. Soil samples were collected from Hilly areas of Muzaffarabad, and oil processing area of Lahore, Pakistan to isolate PHA producing bacteria. The bacteria were screened on Sudan black B and Nile blue A staining methodologies. An efficient PHA producing strain AZR-1 was isolated and characterized morphologically and biochemically. The strain was identified as Bacillus sp. AZR-1 on the genetic basis of 16S rRNA gene sequence. Bacillus sp. AZR-1 produced 40% PHA contents when grown on N-limited mineral salt medium supplemented with glucose in shake flasks. Starch and sodium gluconate were found to be other putative carbon sources for PHA production as the bacterium produced 22 and 17% PHA on these sources. FTIR confirmed the presence of scl-PHA monomers in the polymer extracted from the bacterium. The PHA synthase gene (phaC) was partially amplified and sequenced which showed maximum similarity with the PHA synthase of Bacillus weihenstephanensis KBAB4.