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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

  • Zhang, Ru (College of Chemistry and Chemical Engineering, Hunan Institute of Engineering) ;
  • Huang, Xue-Mei (College of Chemistry and Chemical Engineering, Hunan Institute of Engineering) ;
  • Yan, Hui-Juan (College of Chemistry and Chemical Engineering, Hunan Institute of Engineering) ;
  • Liu, Xin-Yi (College of Chemistry and Chemical Engineering, Hunan Institute of Engineering) ;
  • Zhou, Qi (College of Chemistry and Chemical Engineering, Hunan Institute of Engineering) ;
  • Luo, Zhi-Yong (Molecular Biology Research Center, School of Life Sciences, Central South University) ;
  • Tan, Xiao-Ning (The Affiliated Hospital of Hunan Academy of Chinese Medicine) ;
  • Zhang, Bian-Ling (College of Chemistry and Chemical Engineering, Hunan Institute of Engineering)
  • Received : 2018.08.29
  • Accepted : 2018.11.19
  • Published : 2019.03.28

Abstract

To investigate a novel ${\beta}$-glucosidase from Bifidobacterium breve ATCC 15700 (BbBgl) to produce compound K (CK) via ginsenoside $F_2$ 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, $35^{\circ}C$, 2 or 6 U/ml, and its activity was enhanced by $Mn^{2+}$ significantly. Under the optimal conditions, the half-life of the BbBgl is 180 h, much longer than the characterized ${\beta}$-glycosidases, and the $K_m$ and $V_{max}$ values are 2.7 mM and $39.8{\mu}mol/mg/min$ 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.

Keywords

References

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