2019 ; Vol.29-3: 410~418
|Author||Ru Zhang, Xue-Mei Huang, Hui-Juan Yan, Xin-Yi Liu, Qi Zhou, Zhi-Yong Luo, Xiao-Ning Tan, Bian-Ling Zhang|
|Place of duty||College of Chemistry and Chemical Engineering, Hunan Institute of Engineering, Xiangtan 411104, P. R. China,Hunan Provincial Key Laboratory of EnvironmentaI Catalysis & Waste Recycling, Hunan Institute of Engineering, Xiangtan 411104, P. R. China|
|Title||Highly Selective Production of Compound K from Ginsenoside Rd by Hydrolyzing Glucose at C-3 Glycoside Using β-Glucosidase of Bifidobacterium breve ATCC 15700|
J. Microbiol. Biotechnol.2019 ;
|Abstract||To investigate a novel β-glucosidase from Bifidobacterium breve ATCC 15700 (BbBgl) to produce
compound K (CK) via ginsenoside F2 by highly selective and efficient hydrolysis of the C-3
glycoside from ginsenoside Rd, the BbBgl gene was cloned and expressed in E. coli BL21. The
recombinant BbBgl was purified by Ni-NTA magnetic beads to obtain an enzyme with specific
activity of 37 U/mg protein using pNP-Glc as substrate. The enzyme activity was optimized at
pH 5.0, 35°C, 2 or 6 U/ml, and its activity was enhanced by Mn2+ significantly. Under the
optimal conditions, the half-life of the BbBgl is 180 h, much longer than the characterized
β-glycosidases, and the Km and Vmax values are 2.7 mM and 39.8 μmol/mg/min for ginsenoside
Rd. Moreover, the enzyme exhibits strong tolerance against high substrate concentration (up
to 40 g/l ginsenoside Rd) with a molar biotransformation rate of 96% within 12 h. The good
enzymatic properties and gram-scale conversion capacity of BbBgl provide an attractive
method for large-scale production of rare ginsenoside CK using a single enzyme or a
combination of enzymes|
|Key_word||Biﬁdobacterium breve, biotransformation, compound K, ginsenoside Rd, β-Glucosidase|
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