Bioconversion of ginsenoside Rc into Rd by a novel α-l- arabinofuranosidase, Abf22-3 from Leuconostoc sp. 22-3: cloning, expression, and enzyme characterization

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Title
Bioconversion of ginsenoside Rc into Rd by a novel α-l- arabinofuranosidase, Abf22-3 from Leuconostoc sp. 22-3: cloning, expression, and enzyme characterization
Author(s)
Q M Liu; H M Jung; C H Cui; Bong Hyun Sung; J K Kim; Song-Gun Kim; S T Lee; S C Kim; W T Im
Bibliographic Citation
Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology, vol. 103, no. 4, pp. 747-754
Publication Year
2013
Abstract
A novel α-l-arabinofuranosidase (Abf22-3) that could biotransform ginsenoside Rc into Rd was obtained from the ginsenoside converting Leuconostoc sp. strain 22-3, isolated from the Korean fermented food kimchi. The gene, termed abf22-3, consisting of 1,527 bp and encoding a protein with a predicted molecular mass of 58,486 Da was cloned into the pMAL-c2x (TEV) vector. A BLAST search using the Abf22-3's amino acid sequence revealed significant homology to that of family 51 glycoside hydrolases. The over-expressed recombinant Abf22-3 in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinofuranoside moiety attached to the C-20 position of ginsenoside Rc under optimal conditions of pH 6.0 and 30 C. This result indicated that Abf22-3 selectively converts ginsenoside Rc into Rd, but did not catalyze the hydrolysis of glucopyranosyl groups from Rc or other ginsenosides such as Rb1 and Rb2. Over-expressed recombinant enzymes were purified by two steps with amylose-affinity and DEAE-cellulose chromatography and then characterized. The kinetic parameters for α-l-arabinofuranosidase showed apparent K m and Vmax values of 0.95 ± 0.02 μM and 1.2 ± 0.1 μmol min-1 mg of protein-1 against p-nitrophenyl-α-l-arabinofuranoside, respectively. Using a purified MBP-Abf22-3 (10 μg/ml), 0.1 % of ginsenoside Rc was completely converted to ginsenoside Rd within 20 min.
Keyword
α-l-ArabinofuranosidaseBiotransformationGinsenoside RcLeuconostoc
ISSN
0003-6072
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s10482-012-9856-2
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
Jeonbuk Branch Institute > Biological Resource Center > 1. Journal Articles
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