We describe a high-throughput procedure for measuring β-galactosidase activity in bacteria. This procedure is performed by facultative anaerobic microbial consortium Bacillus safensis. using MMRS medium in batch mode (using Erlenmeyer flasks) and at large-scale (2.0 liter Eyela made Jar fermenter) at different operating parameters namely incubation temperature and stirrer speed for optimizing microbial growth and synthesis of intracellular β-galactosidase. Microbial growth was experimentally determined by biomass concentration, lactose and protein was estimation. Data of optimization of microbial growth kinetics is analyzed by Response surface methodology (RSM), where linear model gave better results than quadratic model in terms of standard deviation. It was brought up that the production route of intracellular β-galactosidase associated with microbial growth, under controlled condition of varying concentration of commercial lactose in Eyela made 2.0 liter Jar fermenter is more economical and unique, leading to a large economical boon which is expected in dairy and biotechnological industries.