Vol. 2, Issue 5 (2017)
Production and partial purification of collagenase from Bacillus sp. isolated from soil sample
Author(s): Aditi Chauhan, Vijay Prabha
Abstract: Collagenases are endopeptidases which are able to digest both native and denatured collagen. The peptides resulting from the collagen degradation by collgenases have wide range of applications and this bioconversion of collagen in to value added digested peptides have attracted a worldwide attention. Hence, the present work was aimed to isolate new collagenase producing microorganisms, optimize various physico chemical parameters for collagenase production and partial purification of the collagenase enzyme. From different soil samples, 20 bacterial isolates showed digestion of gelatin on gelatin agar medium and out of them only four isolates confirmed collagenase production by azocoll. Among these four isolates, isolate Ad18 showed highest collagenase enzyme activity by ninhydrin assay. Ad18 was then characterized for its morphological and biochemical characters. The effect of different parameters like carbon sources, nitrogen sources, temperature, pH, inoculum size and incubation period was monitored with the selected strain for collagenase production. The maximum enzyme production was obtained when the basal media of pH 7.5 containing gelatin (1.5%, w/v), ammonium ferrous sulphate (1%, w/v) was inoculated with 1 % (v/v) inoculum and incubated at 37°C for 72 h. Further, the enzyme was precipitated out with ammonium sulphate at the saturation of 60-80% and partially purified by using Sephadex G-200.