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通过共价异肽键将其锚定在乳酸乳球菌表面

Surface Anchoring on Lactococcus lactis by Covalent Isopeptide Bond.

作者信息

Plavec Tina Vida, Berlec Aleš

出版信息

Acta Chim Slov. 2019 Feb;66(1):18-27.

Abstract

Display of recombinant proteins on the bacterial surface is an emerging research area with wide range of potential biotechnological applications. Because of its GRAS (generally recognized as safe) status, lactic acid bacterium Lactococcus lactis represents an attractive host for surface display and promising vector for in situ delivery of bioactive proteins. The present study focused on finding a new alternative approach for surface display on Lactoccocus lactis. We developed a system that enables the formation of irreversible isopeptide bonds on the surface of Lactococus lactis. This was achieved through the following two protein/peptide pairs, SpyCatcher/SpyTag and SnoopCatcher/SnoopTag.1-3 Attachment of tagged model protein B domain to the cell surface of Lactococccus lactis displaying the corresponding catcher protein was demonstrated using flow cytometry. We demonstrated effective use of aforementioned protein anchors which thus represent a promising alternative to established approaches for surface display on Lactoccocus lactis.

摘要

在细菌表面展示重组蛋白是一个新兴的研究领域,具有广泛的潜在生物技术应用。由于其公认安全(GRAS)的地位,乳酸乳球菌是一种有吸引力的表面展示宿主,也是原位递送生物活性蛋白的有前景的载体。本研究重点在于寻找一种在乳酸乳球菌表面展示的新替代方法。我们开发了一种能够在乳酸乳球菌表面形成不可逆异肽键的系统。这是通过以下两对蛋白质/肽实现的,即SpyCatcher/SpyTag和SnoopCatcher/SnoopTag。1-3 使用流式细胞术证明了将标记的模型蛋白B结构域附着到展示相应捕获蛋白的乳酸乳球菌细胞表面。我们证明了上述蛋白质锚定物的有效应用,因此它们是乳酸乳球菌表面展示既定方法的一种有前景的替代方法。

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