title: Purification and characterization of ginsenoside Ra-hydrolyzing beta-D-xylosidase from Bifidobacterium breve K-110, a human intestinal anaerobic bacterium.
abstract: Beta-D-Xylosidase (EC 3.2.1.37) has been purified from ginsenoside Ra-metabolizing Bifidobacterium breve K-110, which was isolated from human intestinal microflora. beta-D-Xylosidase was purified to apparent homogeneity by a combination of ammonium sulfate precipitation, QAE-cellulose, butyl-toyopearl, hydroxyapatit and Q-Sepharose column chromatographies with the final specific activity of 51.8 micromol/min/mg. Molecular weight of beta-D-xylosidase is 49 kDa by SDS-PAGE and gel filtration, which consisted of a single subunit. beta-D-Xylosidase showed optimal activity at pH 5.0 and 37 degrees C. The purified enzyme was potently inhibited by PCMS. beta-D-Xylosidase acted to the greatest extent on p-nitrophenyl-beta-D-xylopyranoside, followed by ginsenoside Ra1 and ginsenoside Ra2. This enzyme hydrolyzed xylan to xylose, but did not act on p-nitrophenyl-beta-glucopyranoside, p-nitrophenyl-beta-galactopyranoside or p-nitrophenyl-beta-D-fucopyranoside. These findings suggest that this is the first reported purification of ginsenoside-hydrolyzing beta-D-xylosidase from an anaerobic Bifidobacterium sp.
authors.affiliation: College of Pharmacy, Kyung Hee University, Seoul, Korea.
authors.forename: Ho-Young
authors.initials: HY
authors.lastname: Shin
authors.forename: Ji-Hyun
authors.initials: JH
authors.lastname: Lee
authors.forename: Jang-Yeon
authors.initials: JY
authors.lastname: Lee
authors.forename: Yeo-Ock
authors.initials: YO
authors.lastname: Han
authors.forename: Myung Joo
authors.initials: MJ
authors.lastname: Han
authors.forename: Dong-Hyun
authors.initials: DH
authors.lastname: Kim
mesh_terms: D001644:Bifidobacterium / Q000302:isolation & purification* / Q000378:metabolism; D036145:Ginsenosides / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000382:microbiology*; D014995:Xylosidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D036145:Ginsenosides; D014995:Xylosidases; C026579:exo-1,4-beta-D-xylosidase