Screening, optimization of culture conditions and scale-up for production of the L-Glutaminase by novel isolated Bacillus sps. mutant endophyte using response surface methodology

Abstract

Abstract L-Glutaminase is known as the anti-cancer drug to treat anti-leukemic cancer besides its use in food industry for the acrylamide degradation of fried foods. In the present study, submerged fermentation was carried out for the production; statistical based experimental designs were employed to optimize the culture conditions and to maximize the L -Glutaminase activity from novel Bacillus sps., mutant endophyte isolated from Ocimum tenuiflorum. Eleven process variables of the medium were examined for their significance using Plackett-Burman design on the production of L-Glutaminase i.e., Galactose, L -Glutamine, Na2HPO4, KH2PO4, NaCl, MgSO4·H2O, Yeast extract, pH, Temperature, RPM, Time were screened and it was observed that activity was 22.49 IU/ml. After Plackett-Burman design experiments, the activity was found to be 2.34 folds increase compared to one variable at a time method. Further, the production was optimized with Response Surface Methodology (RSM) using Central Composite Design (CCD) for significant variables i.e., Galactose (g/L), Time (h), Temperature (T°C) and pH obtained from Plackett-Burman design and the activity was found to be 24.585 IU/ml i.e., 2.56 fold increment in the production compared to unoptimized medium. Experimentation revealed that the isolated novel Bacillus sps., mutants as the potential bacterial source for L -Glutaminase production.