Screening of polyhydroxybutyrate producing indigenous bacteria from polluted lake soil

Abstract

The prime aim of this study was to enumerate predominant bacteria from polluted lake soil samples, which possess polyhydroxybutyrate (PHB) fabricating potential and identify the suitable growth conditions and nutritional factors for PHB fabrication. From several numbers of bacterial cultures, one culture has the competence to yield PHB, and it was endorsed through Sudan Black B stain, Nile red staining, SEM analysis, and growth in PHB selective media. Under the microscopic observation, the fluorescent cells and polymeric granules were observed in the fluorescent microscope and SEM, respectively. This PHB fabricating isolate was recognized as Bacillus cereus NDRMN001 through 16S rRNA partial sequence analysis. The structural characteristics of PHB produced by B. cereus NDRMN001 were studied through FT-IR, 1H NMR, and 13C NMR analysis. The peak observed at 1759.27 cm−1 on FT-IR analysis is corresponding to the signal band of PHB. In 1H NMR peaks were noticed at 1.67, 2.37 to 2.71, and 3.38 to 7.68 which corresponding to –CH3, –CH2, and –CH protons of PHB. About 4 notable peaks were noticed in 13C NMR analysis at 19.62, 68.27, 40.68, and 169.11 ppm which appeared close to the carboxyl group of PHB. About 10% of inoculum, pH 7.5, 2 g L of yeast extract, 20 g L of rice bran, 35 °C, and 2 days of incubation were recognized as optimal growth conditions for B. cereus NDRMN001 to produce PHB. The identified B. cereus NDRMN001 has the potential to yield 91.48% of PHB as 33.19 g L of PHB from 36.26 g L of culture biomass. The complete results conclude that the B. cereus NDRMN001 screened from polluted lake soil has the competence to produce fine quality and quantity of PHB in a short duration of fabrication process under favorable conditions with the utilization of cheap nutritional factors.