title: Characterization of recombinant β-glucosidase from Arthrobacter chlorophenolicus and biotransformation of ginsenosides Rb1, Rb 2, Rc, and Rd.
abstract: 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.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Republic of Korea.
authors.forename: Myung Keun
authors.initials: MK
authors.lastname: Park
authors.forename: Chang-Hao
authors.initials: CH
authors.lastname: Cui
authors.forename: Sung Chul
authors.initials: SC
authors.lastname: Park
authors.forename: Seul-Ki
authors.initials: SK
authors.lastname: Park
authors.forename: Jin-Kwang
authors.initials: JK
authors.lastname: Kim
authors.forename: Mi-Sun
authors.initials: MS
authors.lastname: Jung
authors.forename: Suk-Chae
authors.initials: SC
authors.lastname: Jung
authors.forename: Sun-Chang
authors.initials: SC
authors.lastname: Kim
authors.forename: Wan-Taek
authors.initials: WT
authors.lastname: Im
mesh_terms: D001173:Arthrobacter / Q000201:enzymology* / Q000235:genetics; D001711:Biotransformation; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D036145:Ginsenosides / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature; D001617:beta-Glucosidase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D036145:Ginsenosides; D011994:Recombinant Proteins; D001617:beta-Glucosidase