Bioprospecting amylase from Samiti Lake, situated in the eastern Himalayas

Enzymes, especially amylases, have been an economic boon to the industrial sector, their bioprospective and biotechnological use is an added advantage. Our primary focus of the study was to isolate the most potent amylase producer and to optimize its production parameters through One Factor At A Time (OFAT), Central Composite Rotatable Design Response Surface Methodology (CCRD RSM) and Artificial Neural Network (ANN). Based on the qualitative and quantitative analysis, SLAB1 was selected as the most potent amylase producer out of the potential isolates. Further SLAB1 was identified as Priestia flexa via 16SrRNA identification. Optimization of the production parameters showed the best carbon, nitrogen sources, temperature and pH to be fructose, peptone, 20 °C and pH 8.0 respectively. Further, the enzyme was purified using ammonium sulphate precipitation followed by dialysis. Later, DEAE Sepharose (Sigma) resin was used for ion exchange chromatography and the protein was eluted using NaCl gradients from 0.1 M - 0.6 M. Enzyme kinetics assessment of the purified amylase with the Lineweaver Burk plot showed values of maximum rate; Vmax (10.869 μmoL/min), and Michaelis-Menten constant Km to be around (14.91 mg/ml). To determine its potential application, analysis of this purified amylase in cleaning the tomato and chocolate stained cotton fabrics after comparing its compatibility with different detergents were executed. Further analysis of the washed stained fabrics via Scanning Electron Microscopy was carried out.