Characterization of a novel ginsenoside-hydrolyzing [alpha]-l-arabinofuranosidase, AbfA, from Rhodanobacter ginsenosidimutans Gsoil 3054^sup T

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-glucopyrano...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2012-05, Vol.94 (3), p.673
Main Authors: An, Dong-shan, Cui, Chang-hao, Sung, Bong Hyun, Yang, Hee-chan, Kim, Sun Chang, Lee, Sung-taik, Im, Wan-taek, Kim, Song-gun
Format: Article
Language:English
Subjects:
Analysis
Bacteria
Biotechnology
Cloning
E coli
Enzymes
Flowers & plants
Genetic recombination
Studies
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Summary: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^sup 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 ^sub m^ values of 0.53±0.07 and 0.30±0.07 mM and V ^sub max^ values of 27.1±1.7 and 49.6±4.1 μmol min^sup -1^ mg^sup -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.[PUBLICATION ABSTRACT]
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-011-3614-7