Sucrose phosphorylase from Lactobacillus reuteri: Characterization and application of enzyme for production of 2-O-α-d-glucopyranosyl glycerol

The enzymatic synthesis of 2-O-α-d-glucopyranosyl-glycerol (2-αGG) by transglycosylation activity of sucrose phosphorylase (SPase) is a promising method for 2-αGG manufacturing. However, there are only a few SPases available for 2-αGG production. Here, we report on the characterization and application of SPase from Lactobacillus reuteri (LrSPase). The results of transglycosylation properties assay showed that LrSPase was a potential glycerol glycosylating tool with high activity at pH 8.0 and 45 °C. And the transglycosylation activity of LrSPase was seriously inhibited by Fe3+, Zn2+ and Cu2+. Moreover, the result of substrate specificity assay showed LrSPase was able to catalyze the transglycosylation of 13 phenolic compounds. To produce commercially relevant concentrations of 2-αGG, we have developed a practical, efficient and scalable process for 2-αGG production using sucrose batch-feeding strategy by whole-cell catalyst. The maximum titer of 2-αGG was 237.68 g L-1 with a productivity of 23.39 mM h-1 and the molar conversion rate of glycerol reached 62.38%. To the best of our knowledge, this is the highest 2-αGG production level by using only SPase to synthesize 2-αGG until now. This study provides an effective way for industrial production of 2-αGG.