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KMID : 0545120190290030410
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Journal of Microbiology and Biotechnology
2019 Volume.29 No. 3 p.410 ~ p.418
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Highly Selective Production of Compound K from Ginsenoside Rd by Hydrolyzing Glucose at C-3 Glycoside Using β-Glucosidase of Bifidobacterium breve ATCC 15700
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Zhang Ru
Huang Xue-Mei
Yan Hui-Juan
Liu Xin-Yi
Zhou Qi
Luo Zhi-Yong
Tan Xiao-Ning
Zhang Bian-Ling
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Abstract
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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.
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KEYWORD
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Bi?dobacterium breve, biotransformation, compound K, ginsenoside Rd, β-Glucosidase
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