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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.
- Authors
Liu, Qing-Mei; Jung, Hae-Min; Cui, Chang-Hao; Sung, Bong-Hyun; Kim, Jin-Kwang; Kim, Song-Gun; Lee, Sung-Taik; Kim, Sun-Chang; Im, Wan-Taek
- 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 Rb and Rb. 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 and V values of 0.95 ± 0.02 μM and 1.2 ± 0.1 μmol min mg of protein 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.
- Subjects
GINSENOSIDES; LEUCONOSTOC; ARABINOFURANOSIDASES; BIOCONVERSION; FERMENTED foods; KIMCHI
- Publication
Antonie van Leeuwenhoek, 2013, Vol 103, Issue 4, p747
- ISSN
0003-6072
- Publication type
Article
- DOI
10.1007/s10482-012-9856-2