title: Display of Fibrobacter succinogenes β-glucanase on the cell surface of Lactobacillus reuteri.
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.
authors.affiliation: Department of Animal Science and Technology and Institute of Biotechnology, National Taiwan University , Taipei, Taiwan.
authors.forename: Shu-Jung
authors.initials: SJ
authors.lastname: Huang
authors.forename: Ming-Ju
authors.initials: MJ
authors.lastname: Chen
authors.forename: Pei-Ying
authors.initials: PY
authors.lastname: Yueh
authors.forename: Bi
authors.initials: B
authors.lastname: Yu
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.forename: Je-Ruei
authors.initials: JR
authors.lastname: Liu
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D018938:Caco-2 Cells; D045848:Fibrobacter / Q000201:enzymology*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D052200:Limosilactobacillus reuteri / Q000201:enzymology*; D018613:Microscopy, Confocal; D010957:Plasmids / Q000235:genetics; D019936:Probiotics; D011993:Recombinant Fusion Proteins; D012417:Rumen / Q000382:microbiology; D047071:beta-Glucans / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D047071:beta-Glucans; D006026:Glycoside Hydrolases