期刊
JOURNAL OF GINSENG RESEARCH
卷 36, 期 4, 页码 418-424出版社
KOREAN SOC GINSENG
DOI: 10.5142/jgr.2012.36.4.418
关键词
Panax ginseng; Biotransformation; beta-glucosidase; Ginsenoside F2; Flavobacterium johnsoniae
资金
- Next-Generation BioGreen 21 Program, Rural Development Administration [PJ008193]
- Intelligent Synthetic Biology Center of Global Frontier Project
- Ministry of Education, Science and Technology, Republic of Korea [2011-0031967]
- National Research Foundation of Korea [2011-0031967, 2011-0031955] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- Rural Development Administration (RDA), Republic of Korea [PJ008193012012] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
This study focused on the enzymatic biotransformation of the major ginsenoside Rb1 into Rd for the mass production of minor ginsenosides using a novel recombinant beta-glucosidase from Flavobacterium johnsoniae. The gene (bglF3) consisting of 2,235 bp (744 amino acid residues) was cloned and the recombinant enzyme overexpressed in Escherichia coli BL21(DE3) was characterized. This enzyme could transform ginsenoside Rb-1 and gypenoside XVII to the ginsenosides Rd and F2, respectively. The glutathione S-transferase (GST) fused BglF3 was purified with GST-bind agarose resin and characterized. The kinetic parameters for beta-glucosidase had apparent K-m values of 0.91 +/- 0.02 and 2.84 +/- 0.05 mM and V-max values of 5.75 +/- 0.12 and 0.71 +/- 0.01 mu mol.min(-1).mgw of protein(-1) against p-nitrophenyl-beta-D-glucopyranoside and Rb1, respectively. At optimal conditions of pH 6.0 and 37 degrees C, BglF3 could only hydrolyze the outer glucose moiety of ginsenoside Rb1 and gypenoside XVII at the C-20 position of aglycon into ginsenosides Rd and F2, respectively. These results indicate that the recombinant BglF3 could be useful for the mass production of ginsenosides Rd and F2 in the pharmaceutical or cosmetic industry.
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