Efficient Biosynthesis of Lactosucrose from Sucrose and Lactose by the Purified Recombinant Levansucrase from Leuconostoc mesenteroides B-512 FMC.

Lactosucrose, a rare trisaccharide formed from sucrose and lactose by enzymatic transglycosylation, is a type of indigestible carbohydrate with a good prebiotic effect. In this study, lactosucrose biosynthesis was efficiently carried out by a purified levansucrase from Leuconostoc mesenteroides B-512. The target gene was cloned and expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity by nickel affinity and gel filtration chromatography. The effects of pH, temperature, substrate concentration, substrate ratio, and enzyme amount on lactosucrose biosynthesis were studied in detail, and the optimized conditions were determined to be pH 6.5, 50 °C, 27% (W/V) sucrose, 27% (W/V) lactose, and 5 U mL(-1) of the purified recombinant enzyme. Under the optimized reaction conditions, the maximal lactosucrose yield reached 224 g L(-1) after reaction for 1 h. Therefore, L. mesenteroides levansucrase could be considered a potential candidate for future industrial production of lactosucrose.