BIOSYNTHESIS AND OPTIMIZATION OF AMYLASE FROM BACILLUS SP. ISOLATED FROM SOIL SAMPLES USING AGROINDUSTRIAL WASTE AS SUBSTRATE

Amylase enzyme is used in various industries due to its diverse applications. In this study, bacteria from soil sample were primarily screened on starch agar medium to identify amylase producer through the detection of prominent clear zone. Total of five soil samples namely bakery points (A-1), sugar cane juice point (A-2), Lichi chinesis garden soil (A-3), rice field (A-4) and sugar industrial waste (A-5) were used in this study. Among the 17 strains isolated from three samples A-1, A-2 and A-3 were found positive for amylase production. The strains were further screened on the production medium. The N-1 bacterial strain revealed higher enzyme activity (92.21 +/- 17 IU/ml) compared to the other strain and was thus selected for further work. The strain was identified as Bacillus lichenoformis from the 16S rRNA analysis. Enzyme production was enhanced by optimizing various parameters by one factor at a time technique. The agro industrial waste rice polish was used as substrate. The optimum temperature of the enzyme was 35 degrees C, pH 5.5 and 2% (w/v) of substrate concentration. Qualitative detection by using sodium dodecyl polyacrylamide gel electrophoresis showed that molecular weight of enzyme was 35 kDa. This indicated that the enzyme requires a moderately high temperature and neutral pH to show greatest activity.