Cloning and sequencing of the gene for cellobiose 2-epimerase from a ruminal strain of Eubacterium cellulosolvens

被引：49

作者：

Taguchi, Hidenori ^{[1
]}

Senoura, Takeshi

Hamada, Shigeki ^{[2
]}

Matsui, Hirokazu ^{[2
]}

Kobayashi, Yasuo ^{[3
]}

Watanabe, Jun

Wasaki, Jun

Ito, Susumu

机构：

[1] Hokkaido Univ, CRIS, Kita Ku, Sapporo, Hokkaido 0010021, Japan

[2] Hokkaido Univ, Dept Appl Biosci, Grad Sch Agr, Sapporo, Hokkaido 0010021, Japan

[3] Hokkaido Univ, Anim Nutr Lab, Grad Sch Agr, Sapporo, Hokkaido 0010021, Japan

来源：

FEMS MICROBIOLOGY LETTERS | 2008年 / 287卷 / 01期

关键词：

Eubacterium; rumen; cellobiose; 2-epimerase; N-acyl-D-glucosamine; epilactose;

D O I：

10.1111/j.1574-6968.2008.01281.x

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Cellobiose 2-epimerase (CE; EC 5.1.3.11) is known to catalyze the reversible epimerization of cellobiose to 4-O-beta-D-glucopyranosyl-D-mannose in Ruminococcus albus cells. Here, we report a CE in it ruminal strain of Eubacterium cellulosolvens for the first time. The nucleotide sequence of the CE had all ORF of 1218 bp (405 amino acids; 46 963.3 Da). The CE front E. cellulosolvens showed 44-54% identify to N-acyl-D-glucosamine 2-epimerase-like hypothetical proteins in the genomes of coprococcus eutactus, Faecalibacterium prausnitzii, Clostridium phytofermentans, Calidecellulosiruptor saccharolyticus, and Eubacterium siraeum. Surprisingly, it exhibited only 46% identify to a CE from R. albus. The recombinant enzyme expressed in Escherichia coli was purified by two-step chromatography. The purified enzyme had a molecular mass of 46.7kDa and exhibited optimal activity at around 35 degrees C and pH 7.0-8.5. In addition to cello-oligosaccharides, it converted lactose to epilactose (4-O-beta-D-galactopyranosyl-D-mannose).

引用

页码：34 / 40

页数：7

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