ABSTRACT
Objective: In this study, gelatinase producing bacteria were probed from sediment samples of Porto Novo Coastal sites, India. Screening and identification of potential strain were done followed by optimization of physico-chemical parameters; bulk production and gelatinase extraction were carried out. Methods: For probing of gelatinase potential producer primary and secondary screening was carried out for qualitative and quantitative estimation. Optimization of physico-chemical parameters for improved production of gelatinase enzyme and large scale of gelatinase was produced. Gelatinase precipitation was standardized using different saturation rates of ammonium sulphate from 10 to 100% at 4℃.Results: There were 8 morphologically different gelatinase producing bacteria were initially delved through primary screening tests. Bacillus spp produced maximum gelatinase activity (2.1U/mL) in secondary screening test. Optimizing its abiotic and biotic factors, maximum enzyme activity was achieved at 48h incubation period (2.2U/mL), 2.5 pH (2.5U/mL), 35℃ temperature (2.55U/mL), 0.8% lactose (2.6U/mL), 1.4% gelatin (2.9U/mL) as the ideal carbon source and nitrogen source, 1% salinity (2.9U/mL) and 3ml of inoculum containing 5.6×106/ mL (3.3U/mL). From the optimized factors, bulk production was carried out and saturation rate of 40% ammonium sulphate, precipitated out maximum enzyme with lowered dry weight indicates its enzyme purity and recovered enzyme showed 4.1U/mg of activity. Conclusion: The study revealed that the isolated strain Bacillus spp has its potentiality for industrial scale production and the results will stand as a base line data for the application of gelatinase in future.