Department of Animal Science and Technology and Institute of Biotechnology, National Taiwan University , Taipei, Taiwan.
J Agric Food Chem. 2011 Mar 9;59(5):1744-51. doi: 10.1021/jf104266x. Epub 2011 Feb 8.
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.
本研究旨在将一种瘤胃细菌β-葡聚糖酶展示在益生菌罗伊氏乳杆菌的细胞表面。评估了遗传修饰菌株的β-葡聚糖降解能力和黏附能力。将纤维丁酸弧菌的β-葡聚糖酶(Glu)融合到罗伊氏乳杆菌 Cnb 的 C 末端,然后由罗伊氏乳杆菌 Pg4 表达为重组 Cnb-Glu-His(6)融合蛋白。转化菌株 L. reuteri pNZ-cnb/glu 的共聚焦免疫荧光显微镜和流式细胞术分析表明,Cnb-Glu-His(6)融合蛋白被展示在其细胞表面。此外,与亲本菌株 L. reuteri Pg4 相比,L. reuteri pNZ-cnb/glu 获得了分解大麦β-葡聚糖的能力,并表现出更高的黏附能力。据作者所知,这是首次成功地将纤维分解酶展示在肠道乳杆菌的细胞表面上。