RRC ID 17400
著者 Ito S, Taguchi H, Hamada S, Kawauchi S, Ito H, Senoura T, Watanabe J, Nishimukai M, Ito S, Matsui H.
タイトル Enzymatic properties of cellobiose 2-epimerase from Ruminococcus albus and the synthesis of rare oligosaccharides by the enzyme.
ジャーナル Appl Microbiol Biotechnol
Abstract The gene for cellobiose 2-epimerase (CE) from Ruminococcus albus NE1 was overexpressed in Escherichia coli cells. The recombinant CE was purified to homogeneity by a simple purification procedure with a high yield of 88%, and the molecular mass was 43.1 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis and 44.0 kDa on gel chromatography. It exhibited optimal activity around at 30 degrees C and pH 7.5, and the enzyme activity was inhibited by Al3+, Fe3+, Co2+, Cu2+, Zn2+, Pb2+, Ag+, N-bromosuccinimide, iodoacetate, and 4-chloromercuribenzoate. In addition to cello-oligosaccharides, the enzyme was found to effectively 2-epimerize lactose to yield 4-O-beta-D-galactopyranosyl-D-mannose (epilactose), which occurs in cow milk as a rare oligosaccharide. The Km and kcat/Km values toward lactose were 33 mM and 1.6 s(-1) mM(-1), and those toward cellobiose were 13.8 mM and 4.6 s(-1) mM(-1), respectively. N-Acetyl-D-glucosamine, uridine 5'-diphosphate-glucose, D-glucose 6-phosphate, maltose, sophorose, laminaribiose, and gentiobiose were inert as substrates for the recombinant CE. We demonstrated that epilactose was resistant to rat intestinal enzymes, utilized by human adult bifidobacteria, and stimulated the tight junction permeability in Caco-2 cells. These results strongly suggest that this rare disaccharide is promising for use as a prebiotic.
巻・号 79(3)
ページ 433-41
公開日 2008-6-1
DOI 10.1007/s00253-008-1449-7
PMID 18392616
MeSH Animals Bacterial Proteins / chemistry* Bacterial Proteins / genetics Bacterial Proteins / isolation & purification Bacterial Proteins / metabolism Bifidobacterium / growth & development Bifidobacterium / metabolism Caco-2 Cells Cattle Cellobiose / metabolism Enzyme Stability Escherichia coli / genetics Escherichia coli / metabolism Gene Expression Humans Kinetics Magnetic Resonance Spectroscopy Molecular Sequence Data Molecular Weight Oligosaccharides / metabolism* Racemases and Epimerases / chemistry* Racemases and Epimerases / genetics Racemases and Epimerases / isolation & purification Racemases and Epimerases / metabolism Rats Ruminococcus / enzymology* Ruminococcus / genetics Ruminococcus / isolation & purification Substrate Specificity Tight Junctions / metabolism
IF 3.53
引用数 51
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
一般微生物 JCM 1192 JCM 1194 JCM 1217 JCM 1255 JCM 1275