Enzymatic Biotransformation of Ginsenoside Rb2 into Rd by Recombinant α-L-Arabinopyranosidase from Blastococcus saxobsidens
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In this study, we used a novel α-L-arabinopyranosidase (AbpBs) obtained from ginsenosideconverting Blastococcus saxobsidens that was cloned and expressed in Escherichia coli BL21 (DE3), and then applied it in the biotransformation of ginsenoside Rb2 into Rd. The gene, termed AbpBs, consisting of 2,406 nucleotides (801 amino acid residues), and with a predicted translated protein molecular mass of 86.4 kDa, was cloned into a pGEX4T-1 vector. A BLAST search using the AbpBs amino acid sequence revealed significant homology with a family 2 glycoside hydrolase (GH2). The over-expressed recombinant AbpBs in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinopyranose moiety attached to the C-20 position of ginsenoside Rb2 under optimal conditions (pH 7.0 and 40°C). Kinetic parameters for α-Larabinopyranosidase showed apparent Km and Vmax values of 0.078 ± 0.0002 μM and 1.4 ± 0.1 μmol/min/mg of protein against p-nitrophenyl-α-L-arabinopyranoside. Using a purified AbpBs (1 μg/ml), 0.1% of ginsenoside Rb2 was completely converted to ginsenoside Rd within 1 h. The recombinant AbpBs could be useful for high-yield, rapid, and low-cost preparation of ginsenoside Rd from Rb2.
Abstract
In this study, we used a novel α-L-arabinopyranosidase (AbpBs) obtained from ginsenosideconverting Blastococcus saxobsidens that was cloned and expressed in Escherichia coli BL21 (DE3), and then applied it in the biotransformation of ginsenoside Rb2 into Rd. The gene, termed AbpBs, consisting of 2,406 nucleotides (801 amino acid residues), and with a predicted translated protein molecular mass of 86.4 kDa, was cloned into a pGEX4T-1 vector. A BLAST search using the AbpBs amino acid sequence revealed significant homology with a family 2 glycoside hydrolase (GH2). The over-expressed recombinant AbpBs in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinopyranose moiety attached to the C-20 position of ginsenoside Rb2 under optimal conditions (pH 7.0 and 40°C). Kinetic parameters for α-Larabinopyranosidase showed apparent Km and Vmax values of 0.078 ± 0.0002 μM and 1.4 ± 0.1 μmol/min/mg of protein against p-nitrophenyl-α-L-arabinopyranoside. Using a purified AbpBs (1 μg/ml), 0.1% of ginsenoside Rb2 was completely converted to ginsenoside Rd within 1 h. The recombinant AbpBs could be useful for high-yield, rapid, and low-cost preparation of ginsenoside Rd from Rb2.
참고문헌 (36)
- Edzard Ernst, "Panax ginseng: An Overview of the Clinical Evidence", Journal of Ginseng Research, 34(4), pp.259~263 (2010), 고려인삼학회
- Mi Hyun Kim, "The Changes of Ginsenoside Patterns in Red Ginseng Processed by Organic Acid Impregnation Pretreatment", Journal of Ginseng Research, 35(4), pp.497~503 (2011), 고려인삼학회
- Yun TK, "Brief introduction of Panax ginseng C .A . Meyer", J. Korean Med. Sci., 16(Suppl), pp.S3~S5 (2001)
- 김현숙, "Effects of Ginsenosides Rg3 and Rh2 on the Proliferation of Prostate Cancer Cells", Archives of Pharmacal Research, 27(4), pp.429~435 (2004), 대한약학회
- Keum YS, "Inhibitory effects of the ginsenoside Rg3 on phorbol ester-induced cyclooxygenase-2 expression, NF-kappaB activation and tumor promotion", Mutat. Res., 523-524, pp.75~85 (2003)
- Kim S, "Neuroprotective effects of ginseng saponins against L-type Ca2+ channel-mediated cell death in rat cortical neurons", Biochem. Biophys. Res. Commun, 365, pp.399~405 (2008)
- Zhang JT, "Preliminary study on antiamnestic mechanism of ginsenoside Rg1 and Rb1", Chin. Med. J, 103, pp.932~938 (1990)
- Attele AS, "Ginseng pharmacology : multiple constituents and multiple actions", Biochem. Pharmacol, 58, pp.1685~1693 (1999)
- Lee TK, "Radioprotective potential of ginseng", Mutagenesis, 20, pp.237~243 (2005)
- 김시관, "Trends in Ginseng Research in 2010", Journal of Ginseng Research, 35(4), pp.389~398 (2011), 고려인삼학회
- Joon-Hee Lee, "Effects of Minor Ginsenosides, Ginsenoside Metabolites, and Ginsenoside Epimers on the Growth of Caenorhabditis elegans", Journal of Ginseng Research, 35(3), pp.375~383 (2011), 고려인삼학회
- Christensen LP, "Chapter 1. Ginsenosides: chemistry, biosynthesis, analysis, and potential health effects", Adv. Food Nutr. Res., 55, pp.1~99 (2009)
- Wang L, "Inhibitory effect of ginsenoside-Rd on carrageenaninduced inflammation in rats", Can. J. Physiol. Pharmacol, 90, pp.229~236 (2012)
- Lin T, "Promotive effect of ginsenoside Rd on proliferation of neural stem cells in vivo and in vitro", J. Ethnopharmacol, 142, pp.754~761 (2012)
- Kim WK, "Wound-healing effect of ginsenoside Rd from leaves of Panax ginseng via cyclic AMP-dependent protein kinase pathway", Eur. J. Pharmacol., 702, pp.285~293 (2013)
- Fei-Liang Zhong, "Cloning and Characterization of Ginsenoside-Hydrolyzing β-Glucosidase from Lactobacillus brevis That Transforms Ginsenosides Rb1 and F2 into Ginsenoside Rd and Compound K", Journal of Microbiology and Biotechnology, 26(10), pp.1661~1667 (2016), 한국미생물·생명공학회[DOI]
- Liu QM, "Bioconversion of ginsenoside Rc into Rd by a novel alpha-L-arabinofuranosidase, Abf22-3 from Leuconostoc sp. 22-3: cloning, expression, and enzyme characterization", Antonie Van Leeuwenhoek, 103, pp.747~754 (2013)
- Yujie Chen, "Determination of ginsenosides in Asian and American ginsengs by liquid chromatographyequadrupole", time-of-flight MS: assessing variations based on morphological characteristics, 2017, pp.41(1), 10~22[DOI]
- Shi WY., "Investigation of ginsenosides in different parts and ages of Panax ginseng", Food Chem, 102, pp.664~668 (2007)
- Li L, "Production of ginsenoside F2 by using Lactococcus lactis with enhanced expression of beta-glucosidase gene from Paenibacillus mucilaginosus", J. Agric. Food Chem, 64, pp.2506~2512 (2016)
- Du J, "Identification and characterization of a ginsenoside-transforming beta-glucosidase from Pseudonocardia sp. Gsoil 1536 and its application for enhanced production of minor ginsenoside Rg2(S)", PLoS One, 9, pp.e96914~ (2014)
- 송봉규, "Production of the Rare Ginsenoside Rh2-MIX (20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3) by Enzymatic Conversion Combined with Acid Treatment and Evaluation of Its Anti-Cancer Activity", Journal of Microbiology and Biotechnology, 27(7), pp.1233~1241 (2017), 한국미생물·생명공학회[DOI]
- Park CS, "Biotransformation of ginsenosides by hydrolyzing the sugar moieties of ginsenosides using microbial glycosidases", Appl. Microbiol. Biotechnol, 87, pp.9~19 (2010)
- An DS, "Identification and characterization of a novel Terrabacter ginsenosidimutans sp. nov. beta-glucosidase that transforms ginsenoside Rb1 into the rare gypenosides XVII and LXXV", Appl. Environ. Microbiol., 76, pp.5827~5836 (2010)
- An DS, "Characterization of a novel ginsenoside-hydrolyzing alpha-L-arabinofuranosidase, AbfA, from Rhodanobacter ginsenosidimutans Gsoil 3054T", Appl. Microbiol. Biotechnol, 94, pp.673~682 (2012)
- Cleland WW, "Statistical analysis of enzyme kinetic data", Methods Enzymol, 63, pp.103~138 (1979)
- Yang M, "Characterization of endo-β-mannanase from Enterobacter ludwigii MY271 and application in pulp industry", Bioprocess Biosyst. Eng, 40, pp.35~43 (2017)
- 박명근, "Characterization of Recombinant β-Glucosidase from Arthrobacter chlorophenolicus and Biotransformation of Ginsenosides Rb1, Rb2, Rc, and Rd", The Journal of Microbiology, 52(5), pp.399~406 (2014), 한국미생물학회[DOI]
- Prajapati BP, "Characterization of cellulase from Aspergillus tubingensis NKBP-55 for generation of fermentable sugars from agricultural residues", Bioresour. Technol, 250, pp.733~740 (2018)
- Salama P, "Photocatalytic oxidation of Ni-EDTA in a well-mixed reactor", Ind. Eng. Chem. Res, 44, pp.7071~7077 (2005)
- Pitter P, "Biodegradability of ethylenediamine-based complexing agents and related compounds", Chemosphere, 44, pp.823~826 (2001)
- 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)
- Cui CH, "Characterization of a ginsenoside-transforming beta-glucosidase from Paenibacillus mucilaginosus and its application for enhanced production of minor ginsenoside F(2)", PLoS One, 9, pp.e85727~ (2014)
- Kim JK, "Mass production of the ginsenoside Rg3(S)through the combinative use of two glycoside hydrolases", Food Chem, 141, pp.1369~1377 (2013)
- Lingli Jiang, "Production of bioactive ginsenoside Rg3(S) and compound K using recombinant Lactococcus lactis", Journal of Ginseng Research, 42(4), pp.412~418 (2018), 고려인삼학회[DOI]
- Wang L, "Bioconversion of ginsenosides Rb1, Rb2, Rc and Rd by novel β-glucosidase hydrolyzing outer 3-O glycoside from Sphingomonas sp. 2F2: cloning, expression, and enzyme characterization", J. Biotechnol., 156, pp.125~133 (2011)