PMID: 8988633Dec 1, 1996Paper

Cloning and nucleotide sequence of the beta-D-glucosidase gene from Bifidobacterium breve clb, and expression of beta-D-glucosidase activity in Escherichia coli

Bioscience, Biotechnology, and Biochemistry
N NunouraH Kumagai

Abstract

Genomic DNA encoding a beta-D-glucosidase (EC 3.2.1.21), which has beta-D-fucosidase activity, was cloned from Bifidobacterium breve clb. We sequenced a 1.9-kbp cloned DNA fragment that contained a single open reading frame encoding 460 amino acids with a calculated molecular mass of 51,513 Da. A putative ribosome binding site was found 5 bp upstream of the initiation codon. The amino acid sequence of this beta-D-glucosidase from Bifidobacterium breve clb had 46% identity with that of beta-glucosidase from Microbispore bispore. The enzyme of Bifidobacterium breve clb was expressed in Escherichia coli. A cell-free extract prepared from the recombinant strain showed 80 to 90-fold more beta-D-glucosidase activity than that from Bifidobacterium breve clb. The recombinant enzyme was purified to homogeneity from cell-free extracts of the recombinant strain using 4 column chromatographies. The recovery of enzyme from the recombinant strain was about 138-fold-higher than that of Bifidobacterium breve clb. The enzymatic properties were similar to those of Bifidobacterium breve clb. For application of this recombinant enzyme, we attempted to synthesize a disaccharide that seemed to be specifically assimilated by Bifidobacteria using the ...Continue Reading

Citations

Jun 1, 2000·Applied and Environmental Microbiology·H TanakaI Mierau
Feb 26, 2005·Critical Reviews in Food Science and Nutrition·Claude P ChampagneDenis Roy
Oct 20, 2005·Journal of Bioscience and Bioengineering·Takane KatayamaKenji Yamamoto
Jul 3, 2015·Frontiers in Microbiology·Taku UchiyamaKentaro Miyazaki
Aug 26, 2016·Applied Microbiology and Biotechnology·Tomohiko Matsuzawa, Katsuro Yaoi
Sep 11, 2003·Biotechnology and Bioengineering·Antonio González VaraDiego Matteuzzi
Jun 28, 2002·Journal of Dairy Science·L LamoureuxS F Gauthier
Nov 28, 2007·Molecular Nutrition & Food Research·Lambertus A M van den BroekAlphons G J Voragen
Sep 21, 2016·International Journal of Molecular Sciences·Seockmo KuHyun Ju You
Jan 11, 2011·Applied and Environmental Microbiology·Karina PokusaevaDouwe van Sinderen
Apr 13, 2012·Biotechnology for Biofuels·Fernando SegatoRolf A Prade

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