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http://dx.doi.org/10.4014/jmb.1910.10065

Enzymatic Biotransformation of Ginsenoside Rb2 into Rd by Recombinant α-L-Arabinopyranosidase from Blastococcus saxobsidens  

Kim, Ju-Hyeon (Department of Biotechnology, Hankyong National University)
Oh, Jung-Mi (Department of Physiology, Chonbuk National University Medical School)
Chun, Sungkun (Department of Physiology, Chonbuk National University Medical School)
Park, Hye Yoon (National Institute of Biological Resources)
Im, Wan Taek (Department of Biotechnology, Hankyong National University)
Publication Information
Journal of Microbiology and Biotechnology / v.30, no.3, 2020 , pp. 391-397 More about this Journal
Abstract
In this study, we used a novel α-L-arabinopyranosidase (AbpBs) obtained from ginsenoside-converting 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℃). Kinetic parameters for α-L-arabinopyranosidase 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 ㎍/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.
Keywords
Ginsenoside $Rb_2$; ginsenoside Rd; biotransformation; ${\alpha}$-L-arabinopyranosidase; Blastococcus saxobsidens;
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