2-O-α-D-Glucosyl glycerol (2-αGG) can be used as a multipurpose anti-aging, cell-stimulating, and skin moisturizing agent in the cosmetic industry. Sucrose phosphorylase (SPase) has been widely used in the production of 2-αGG. In this paper, the gene encoding sucrose phosphorylase from Bifidobacterium longum (BlSP) was inserted into pRSF-Duet-1 to construct the recombinant plasmid pRSF-BlSP and was functionally expressed in E. coli BL21(DE3) to be used as a biocatalyst for the synthesis of 2-αGG firstly. The mutations of BlSP were carried out based on alanine scanning, and a positive mutant G293A with a 50% increase in activity for 2-αGG production was identified. Mutant G293A has less K_m and bigger k_cat/K_m towards glycerol than the parental BlSP. Subsequently, the production of 177.6 g/L 2-αGG was attained from 1 M sucrose and 1.2 M glycerol catalyzed by 17 mg/mL G293A mutant. This study indicated that BlSP has good potential in the production of 2-αGG.

2-O-α-D-Glucosyl glycerol (2-αGG) 在化妆品行业可用作多用途抗衰老、细胞刺激和皮肤保湿剂。蔗糖磷酸化酶 (SPase) 已广泛用于生产 2-αGG。本文将长双歧杆菌（BlSP）编码蔗糖磷酸化酶基因插入 pRSF-Duet-1 构建重组质粒 pRSF-BlSP 并在大肠杆菌BL21（DE3）中功能表达，用作生物催化剂首先合成2-αGG。BlSP 的突变是基于丙氨酸扫描进行的，并确定了一个阳性突变体 G293A，其 2-αGG 产生的活性增加了 50%。突变体 G293A 具有较小的K _m和较大的k _cat /K _m对甘油比亲本 BlSP。随后，由 17 mg/mL G293A 突变体催化的 1 M 蔗糖和 1.2 M 甘油产生 177.6 g/L 2-αGG。该研究表明BlSP在生产2-αGG方面具有良好的潜力。