European Journal of Experimental Biology Open Access

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Isolation and characterization of polyhydroxybutyrate (PHB) producing Bacillus species from agricultural soil

Gurubasappa. G. Biradar, Shivasharana C. T. and Basappa. B. Kaliwal

Polyhydroxyalkanoates (PHAs), which is produced by several bacteria, is a biodegradable polymer that has many industrial and medical applications. The present study deals with the isolation, and characterization of biopolymer (PHB) producing bacteria from the agricultural soil. The biopolymer producing bacteria was isolated from soil samples like crop, nursery, and fodder fields. The bacteria were screened by morphological and biochemical characteristics. The bacteria were further identified by Sudan black- B staining and molecular characterization. High and efficient PHB accumulation bacteria was selected by quantification and extraction method. Further bacteria were confirmed by FTIR analysis to know the functional groups present in the bacteria. A total of six bacillus species were isolated, the strain BBKGBS12 and BBKGBS6 are gram positive and other isolated bacteria were gram negative. Black colour granulesof bacteria BBKGBS6 were seen by Sudan Black-B staining. Also granules of isolated bacteria were fluoroscenced by acridine orange staining . BBKGBS6 identified to be a strain of Lysinibacillus sphaericus BBKGBS6 by 16S r- RNA sequencing and high PHB accumulation. FTIR analysis gave prominent functional groups like CH3, CH2, C=O, C-O, CH, and OH were identified and the results revealed that the poly-3-hydroxybutyrate compound was produced by Lysinibacillus sphaericus BBKGBS6 (Accession number KP403811). However the nursery field soil has showed high amount of PHB accumulation bacteria.