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

FEMS Microbiology Letters
Hidenori TaguchiSusumu Ito

Abstract

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 a ruminal strain of Eubacterium cellulosolvens for the first time. The nucleotide sequence of the CE had an ORF of 1218 bp (405 amino acids; 46 963.3 Da). The CE from E. cellulosolvens showed 44-54% identity to N-acyl-D-glucosamine 2-epimerase-like hypothetical proteins in the genomes of Coprococcus eutactus, Faecalibacterium prausnitzii, Clostridium phytofermentans, Caldicellulosiruptor saccharolyticus, and Eubacterium siraeum. Surprisingly, it exhibited only 46% identity 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 46.7 kDa 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).

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Citations

Aug 22, 2009·Applied Microbiology and Biotechnology·Susumu Ito
Jan 16, 2013·Applied Microbiology and Biotechnology·Wanmeng MuBo Jiang
Feb 10, 2009·Bioscience, Biotechnology, and Biochemistry·Takeshi SenouraSusumu Ito
Nov 12, 2013·Journal of Dairy Science·Manuel KrewinkelLutz Fischer
Jan 8, 2013·Bioscience, Biotechnology, and Biochemistry·Teruyo OjimaHirokazu Matsui
Aug 12, 2010·Bioscience, Biotechnology, and Biochemistry·Wataru SaburiHirokazu Matsui
Dec 10, 2014·Applied Microbiology and Biotechnology·Jun WasakiSusumu Ito
May 5, 2011·Biochemical and Biophysical Research Communications·Takeshi SenouraSusumu Ito
Nov 23, 2016·Journal of the Science of Food and Agriculture·Qiuming ChenWanmeng Mu
Mar 27, 2019·Applied Microbiology and Biotechnology·Yaqin XiaoWanmeng Mu
Jul 21, 2020·Journal of Dairy Science·Qiuming ChenWanmeng Mu
Nov 3, 2020·Acta Crystallographica. Section D, Structural Biology·Yinghui FengRuijin Yang

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