Characterization of recombinant β-glucosidase from Arthrobacter chlorophenolicus and biotransformation of ginsenosides Rb₁, Rb ₂, Rc, and Rd

• Published in: The journal of microbiology 2014-05, Vol.52 (5), p.399-406
• Main Authors: Park, Myung Keun, Cui, Chang-Hao, Park, Sung Chul, Park, Seul-Ki, Kim, Jin-Kwang, Jung, Mi-Sun, Jung, Suk-Chae, Kim, Sun-Chang, Im, Wan-Taek
• Format: Article
• Language: English
• Subjects: agarose; amino acids; Arthrobacter - enzymology; Arthrobacter - genetics; Arthrobacter chlorophenolicus; beta-glucosidase; beta-Glucosidase - genetics; beta-Glucosidase - isolation & purification; beta-Glucosidase - metabolism; Biotransformation; Chromatography, Affinity; Cloning, Molecular; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression; gene overexpression; genes; ginsenosides; Ginsenosides - metabolism; glucose; Hydrogen-Ion Concentration; Hydrolysis; industry; Kinetics; microbiology; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Temperature
• ISSN: 1225-8873; 1976-3794
• DOI: 10.1007/s12275-014-3601-7

The focus of this study was the cloning, expression, and characterization of recombinant ginsenoside hydrolyzing β-glucosidase from Arthrobacter chlorophenolicus with an ultimate objective to more efficiently bio-transform ginsenosides. The gene bglAch, consisting of 1,260 bp (419 amino acid residues) was cloned and the recombinant enzyme, overexpressed in Escherichia coli BL21 (DE3), was characterized. The GST-fused BglAch was purified using GST·Bind agarose resin and characterized. Under optimal conditions (pH 6.0 and 37°C) BglAch hydrolyzed the outer glucose and arabinopyranose moieties of ginsenosides Rb1 and Rb2 at the C20 position of the aglycone into ginsenoside Rd. This was followed by hydrolysis into F₂ of the outer glucose moiety of ginsenoside Rd at the C3 position of the aglycone. Additionally, BglAch more slowly transformed Rc to F₂ via C-Mc₁ (compared to hydrolysis of Rb₁ or Rb₂). These results indicate that the recombinant BglAch could be useful for the production of ginsenoside F₂ for use in the pharmaceutical and cosmetic industries.