Display of Fibrobacter succinogenes β-glucanase on the cell surface of Lactobacillus reuteri

J Agric Food Chem. 2011 Mar 9;59(5):1744-51. doi: 10.1021/jf104266x. Epub 2011 Feb 8.

Abstract

The aim of this study was to display a rumen bacterial β-glucanase on the cell surface of a probiotic Lactobacillus reuteri strain. The β-glucan degrading ability and the adhesion capability of the genetically modified strain were evaluated. The β-glucanase (Glu) from Fibrobacter succinogenes was fused to the C-terminus of collagen-binding protein (Cnb) from L. reuteri and then expressed by L. reuteri Pg4 as a recombinant Cnb-Glu-His(6) fusion protein. Confocal immunofluorescence microscopy and flow cytometric analysis of the transformed strain L. reuteri pNZ-cnb/glu demonstrated that Cnb-Glu-His(6) fusion protein was displayed on its cell surface. In addition, L. reuteri pNZ-cnb/glu acquired the capacity to break down barley β-glucan and showed higher adhesion capability, in comparison with the parental strain L. reuteri Pg4. To the best of the authors' knowledge, this is the first report of successful display of fibrolytic enzymes on the cell surface of intestinal lactobacilli.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Adhesion
  • Caco-2 Cells
  • Fibrobacter / enzymology*
  • Fluorescent Antibody Technique
  • Gene Expression*
  • Glycoside Hydrolases / genetics*
  • Glycoside Hydrolases / metabolism
  • Humans
  • Limosilactobacillus reuteri / enzymology*
  • Microscopy, Confocal
  • Plasmids / genetics
  • Probiotics
  • Recombinant Fusion Proteins
  • Rumen / microbiology
  • beta-Glucans / metabolism

Substances

  • Recombinant Fusion Proteins
  • beta-Glucans
  • Glycoside Hydrolases