Enhancing thermostability and activity of sucrose phosphorylase for high-level production of 2-O-?-d-glucosylglycerol

Sucrose phosphorylase (SPase) can transfer the glucosyl group of sucrose to different compounds and has been widely used in industry. To overcome low thermostability from Leuconostoc mesenteroides ATCC 12291 (LmSP), a method named PROSS was construct mutants with increased thermostability. All variants were screened by measuring their residual activities after heating at 50°C. Then, single point mutant combined improved activity obtained. The half-lives 50°C approximately twice as high those wild type. In addition, 2-O-?-d-glucosylglycerol (?GG) synthesized type two variants, reaction conditions optimized. Under glycerol concentration 3.2 mol/L, 1.2 enzyme 40 U/mL 37°C for 60 h, yield ?GG reached maximum, conversion rate type, V23L V23L/S424R 62.3%, 70.7% 76.3%, respectively. this study, SPase higher stability obtained, achieved high-level production ?GG.