Cloning and Characterization of Ginsenoside-Hydrolyzing β-Glucosidase from Lactobacillus brevis That Transforms Ginsenosides Rb1 and F2 into Ginsenoside Rd and Compound K
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The ginsenoside-hydrolyzing β-glucosidase gene (bgy2) was cloned from Lactobacillus brevis. We expressed this gene in Escherichia coli BL21(DE3), isolated the resulting protein, and then utilized the enzyme for the biotransformation of ginsenosides. The bgy2 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, 30oC), 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.
Abstract
The ginsenoside-hydrolyzing β-glucosidase gene (bgy2) was cloned from Lactobacillus brevis. We expressed this gene in Escherichia coli BL21(DE3), isolated the resulting protein, and then utilized the enzyme for the biotransformation of ginsenosides. The bgy2 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, 30oC), 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.
참고문헌 (21)
- An DS, "Identification and characterization of a novel Terrabacter ginsenosidimutans sp. nov. β-glucosidase that transforms ginsenoside Rb1 into the rare gypenosides XVII and LXXV", Appl. Environ. Microbiol, 76, pp.5827~5836 (2010)
- Bae EA, "Metabolism of ginsenoside Rc by human intestinal bacteria and its related antiallergic activity", Biol. Pharm. Bull, 25, pp.743~747 (2002)
- 배은아, "Transformation of Ginseng Saponins to Ginsenoside Rh2 by Acids and Human Intestinal Bacteria and Biological Activities of Their Transformants", Archives of Pharmacal Research, 27(1), pp.61~67 (2004), 대한약학회
- Chae S, "Effect of compound K, a metabolite of ginseng saponin, combined with gamma-ray radiation in human lung cancer cells in vitro and in vivo", J. Agric. Food Chem, 57, pp.5777~5782 (2009)
- CHENG, LE-QIN, "Microbial Conversion of Ginsenoside Rb1 to Minor Ginsenoside F2 and Gypenoside XVII by Intrasporangium sp. GS603 Isolated from Soil", Journal of Microbiology and Biotechnology, 17(12), pp.1937~1943 (2007), 한국미생물·생명공학회
- Kim JK, "Mass production of the ginsenoside Rg3(S) through the combinative use of two glycoside hydrolases", Food Chem, 141, pp.1369~1377 (2013)
- Myung Kyum Kim, "Microbial Conversion of Major Ginsenoside Rb1 to Pharmaceutically Active Minor Ginsenoside Rd", The Journal of Microbiology, 43(5), pp.456~462 (2005), 한국미생물학회
- Kim WY, "Steaming of ginseng at high temperature enhances biological activity", J. Nat. Prod, 63, pp.1702~1704 (2000)
- So Jin Lee, "Changes in the ginsenoside content during the fermentation process using microbial strains", Journal of Ginseng Research, 39(4), pp.392~397 (2015), 고려인삼학회
- Lin F, "Efficient biotransformation of ginsenoside Rb1 to Rd by isolated Aspergillus versicolor, excreting β-glucosidase in the spore production phase of solid culture", Antonie Van Leeuwenhoek, 108, pp.1117~1127 (2015)
- Ni HX, "The study of ginsenoside on PPARgamma expression of mononuclear macrophage in type 2 diabetes", Mol. Biol. Rep, 37, pp.2975~2979 (2010)
- Park CS, "Biotransformation of ginsenosides by hydrolyzing the sugar moieties of ginsenosides using microbial glycosidases", Appl. Microbiol. Biotechnol, 87, pp.9~19 (2010)
- Quan LH, "Microbial transformation of ginsenoside Rb1 to compound K by Lactobacillus paralimentarius", World J. Microbiol. Biotechnol, 29, pp.1001~1007 (2013)
- 전림호, "Lactobacillus brevis LH8이 생산하는 효소에 의한 Ginsenoside Rd의 Compound K로의 전환", Journal of Ginseng Research, 32(3), pp.226~231 (2008), 고려인삼학회
- Quan LH, "Enzymatic biotransformation of ginsenoside Rb1 to 20(S)-Rg3 by recombinant β-glucosidase from Microbacterium esteraromaticum", Appl. Microbiol. Biotechnol, 94, pp.377~384 (2012)
- Quan LH, "Bioconversion of ginsenoside Rb1 into compound K by Leuconostoc citreum LH1 isolated from kimchi", Braz. J. Microbiol, 42, pp.1227~1237 (2011)
- Quan LH, "Isolation and characterization of novel ginsenosidehydrolyzing glycosidase from Microbacterium esteraromaticum that transforms ginsenoside Rb2 to rare ginsenoside 20(S)-Rg3", Antonie Van Leeuwenhoek, 104, pp.129~137 (2013)
- Wu JY, "Saponin adjuvant enhancement of antigenspecific immune responses to an experimental HIV-1 vaccine", J. Immunol, 148, pp.1519~1525 (1992)
- Xie J, "Characterization of a novel arabinose-tolerant α-Larabinofuranosidase with high ginsenoside Rc to ginsenoside Rd bioconversion productivity", J. Appl. Microbiol, 120, pp.647~660 (2016)
- Xu QF, "Pharmacokinetics and bioavailability of ginsenoside Rb1 and Rg1 from Panax notoginseng in rats", J. Ethnopharmacol, 84, pp.187~192 (2003)
- Yoshikawa M, "Structures of new dammarane-type triterpene saponins from the flower buds of Panax notoginseng and hepatoprotective effects of principal ginseng saponins", J. Nat. Prod, 66, pp.922~927 (2003)