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Display of Fibrobacter succinogenes β-Glucanase on the Cell Surface of Lactobacillus reuteri

机译:罗伊氏乳杆菌细胞表面上琥珀酸纤维原酶β-葡聚糖酶的展示

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摘要

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- His6 fusion protein. Confocal immunofluorescence microscopy and flow cytometric analysis of the transformed strain L. reuteri pNZ-cnb/glu demonstrated that Cnb-Glu-His6 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.
机译:这项研究的目的是在益生菌罗伊氏乳杆菌菌株的细胞表面展示瘤胃细菌β-葡聚糖酶。评价了转基因菌株的β-葡聚糖降解能力和粘附能力。将来自琥珀酸纤维杆菌的β-葡聚糖酶(Glu)融合到来自罗伊氏乳杆菌的胶原结合蛋白(Cnb)的C末端,然后由罗伊氏乳杆菌Pg4表达为重组Cnb_Glu-His6融合蛋白。转化菌株罗伊氏乳杆菌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-His6融合蛋白显示在其细胞表面。另外,与罗伊氏乳杆菌Pg4相比,罗伊氏乳杆菌pNZ-cnb / glu具有分解大麦β-葡聚糖的能力并显示出更高的粘附能力。据作者所知,这是首次在肠道乳酸菌细胞表面成功展示出纤溶酶的报道。

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