Isolation of Novel Bacteria Isolate from Soil for Production of Extra-Cellular Laccase Enzyme

This study reports the isolation of novel Bacterium from Soil for production of Extra-cellular Laccase Enzyme. Samples from garden, ant-induced and animal dung contaminated soil were collected in the Devi Ahilya University campus and around, Indore, Madhya Pradesh. Various methods including biochemical and morphological identification for high productivity of the enzyme were used. The media optimization was implemented taking various parameters like carbon source, nitrogen source, pH and temperature. From the study the bacteria isolate was found to be in Streptomycetes sp. Plates were incubated at 30◦C for 24–72 hours and 10 positive colonies were chosen and purified in stepwise techniques for their potential to produce laccase using 2, 20-azino-bis-[3-ethyl benzothiazoline-6-sulfonic acid] (ATBS) as a substrate. The ant induced soil was found to be the potential source for laccase producing bacteria. Maximum enzyme production was obtained in the presence of starch (0.563U/ml) and NaNO3 (0.84U/ml). The optimum pH and temperature for enzyme activity was found to be at pH 6(0.90U/ml) and temperature at35 oC (0.504U/ml). Soluble starch was used as the substrate for the amylase production. The protocol developed in our study for the isolation, characterization and media optimization for the production of laccase from the isolate is reproducible for further study and industrial applications. Keywords-Isolates, laccase, screening, Streptomycetes sp.