Zhong Fei-Liang, Ma Rui, Jiang Mingliang, Dong Wei-Wei, Jiang Jun, Wu Songquan, Li Donghao, Quan Lin-Hu
Key Laboratory of Natural Resources of Changbai Mountain and Functional Molecules, Ministry of Education, and Department of Chemistry, College of Science, Yanbian University, Yanji 133002, P.R. China.
J Microbiol Biotechnol. 2016 Oct 28;26(10):1661-1667. doi: 10.4014/jmb.1605.05052.
The ginsenoside-hydrolyzing β-glucosidase gene (2) was cloned from . We expressed this gene in BL21(DE3), isolated the resulting protein, and then utilized the enzyme for the biotransformation of ginsenosides. The 2 gene contains 2,223 bp, and encodes a protein of 741 amino acids that is a member of glycosyl hydrolase family 3. β-Glucosidase (Bgy2) cleaved the outer glucose moieties of ginsenosides at the C-20 position, and the inner glucose at the C-3 position. Under optimal conditions (pH 7.0, 30°C), we used 0.1 mg/ml Bgy2 in 20 mM sodium phosphate buffer (PBS) for enzymatic studies. In these conditions, 1.0 mg/ml ginsenoside Rb1 and ginsenoside F2 were converted into 0.59 mg/ml ginsenoside Rd and 0.72mg/ml compound K, with molar conversion productivities of 69% and 91%, respectively. In pharmaceutical and commercial industries, this recombinant Bgy2 would be suitable for producting ginsenoside Rd and compound K.
人参皂苷水解β-葡萄糖苷酶基因(2)是从……克隆而来。我们在BL21(DE3)中表达了该基因，分离出所得蛋白质，然后将该酶用于人参皂苷的生物转化。该基因包含2223个碱基对，编码一个由741个氨基酸组成的蛋白质，它是糖基水解酶家族3的成员。β-葡萄糖苷酶(Bgy2)在C-20位置切割人参皂苷的外部葡萄糖部分，在C-3位置切割内部葡萄糖。在最佳条件下(pH 7.0，30°C)，我们在20 mM磷酸钠缓冲液(PBS)中使用0.1 mg/ml Bgy2进行酶学研究。在这些条件下，1.0 mg/ml人参皂苷Rb1和人参皂苷F2分别转化为0.59 mg/ml人参皂苷Rd和0.72 mg/ml化合物K，摩尔转化率分别为69%和91%。在制药和商业行业中，这种重组Bgy2将适合生产人参皂苷Rd和化合物K。
