title: Characterization of a novel ginsenoside-hydrolyzing α-L-arabinofuranosidase, AbfA, from Rhodanobacter ginsenosidimutans Gsoil 3054T.
abstract: The gene encoding an α-L-arabinofuranosidase that could biotransform ginsenoside Rc {3-O-[β-D-glucopyranosyl-(1-2)-β-D-glucopyranosyl]-20-O-[α-L-arabinofuranosyl-(1-6)-β-D-glucopyranosyl]-20(S)-protopanaxadiol} to ginsenoside Rd {3-O-[β-D-glucopyranosyl-(1-2)-β-D-glucopyranosyl]-20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol} was cloned from a soil bacterium, Rhodanobacter ginsenosidimutans strain Gsoil 3054(T), and the recombinant enzyme was characterized. The enzyme (AbfA) hydrolyzed the arabinofuranosyl moiety from ginsenoside Rc and was classified as a family 51 glycoside hydrolase based on amino acid sequence analysis. Recombinant AbfA expressed in Escherichia coli hydrolyzed non-reducing arabinofuranoside moieties with apparent K (m) values of 0.53 ± 0.07 and 0.30 ± 0.07 mM and V (max) values of 27.1 ± 1.7 and 49.6 ± 4.1 μmol min(-1) mg(-1) of protein for p-nitrophenyl-α-L-arabinofuranoside and ginsenoside Rc, respectively. The enzyme exhibited preferential substrate specificity of the exo-type mode of action towards polyarabinosides or oligoarabinosides. AbfA demonstrated substrate-specific activity for the bioconversion of ginsenosides, as it hydrolyzed only arabinofuranoside moieties from ginsenoside Rc and its derivatives, and not other sugar groups. These results are the first report of a glycoside hydrolase family 51 α-L-arabinofuranosidase that can transform ginsenoside Rc to Rd.
authors.affiliation: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea.
authors.forename: Dong-Shan
authors.initials: DS
authors.lastname: An
authors.forename: Chang-Hao
authors.initials: CH
authors.lastname: Cui
authors.forename: Bong Hyun
authors.initials: BH
authors.lastname: Sung
authors.forename: Hee-Chan
authors.initials: HC
authors.lastname: Yang
authors.forename: Sun Chang
authors.initials: SC
authors.lastname: Kim
authors.forename: Sung-Taik
authors.initials: ST
authors.lastname: Lee
authors.forename: Wan-Taek
authors.initials: WT
authors.lastname: Im
authors.forename: Song-Gun
authors.initials: SG
authors.lastname: Kim
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D036145:Ginsenosides / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012988:Soil Microbiology; D013379:Substrate Specificity; D044166:Xanthomonadaceae / Q000201:enzymology* / Q000235:genetics*
chemical_list: D036145:Ginsenosides; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase