Investigation of the cell disruption methods for maximizing the extraction of arginase from mutant Bacillus licheniformis (M09) using statistical approach

摘要

Arginase, an intracellular enzyme produced by Bacillus licheniformis (NRS-1264) is effectively used as a drug in the treatment of arginine-dependent cancers, and it is essential for controlling acute neurological disorders. We investigated the effect of various cell disruption methods for maximizing the extraction of intracellular arginase from mutant Bacillus licheniformis (M09), followed by comparing optimization methods, one factor at a time (OFAT), evolutionary operation (EVOP) and response surface method (RSM). Ultrasonication for 2-5 min having a suspension volume in the range of 12-20 mL at a radio frequency power between 30-70 W appeared to be the most effective extraction technique for arginase. The arginase yield decreased in the range of 50-70 W of RF power/16-20 mL suspension volume and 4-5 min sonication time. EVOP predicted a maximum arginase extraction of 3,910 IUL-1 at 2 min sonication having 16 mL suspension volume at 30W RF power. However, response surface optimization suggested an optimized condition of 3 min sonication having 14.5 mL suspension volume at 35W RF power in which the maximum arginase activity in the medium was 3,600 IUL-1.