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肉葡萄球菌:从起始培养物到蛋白质工程平台。

Staphylococcus carnosus: from starter culture to protein engineering platform.

机构信息

Division of Protein Technology, School of Biotechnology, KTH-Royal Institute of Technology, AlbaNova University Center, Roslagstullsbacken 21, 106 91, Stockholm, Sweden.

Microbial Genetics, Interfaculty Institute of Microbiology and Infection Medicine and Infection Medicine (IMIT), University of Tübingen, Auf der Morgenstelle 28, 72076, Tübingen, Germany.

出版信息

Appl Microbiol Biotechnol. 2017 Dec;101(23-24):8293-8307. doi: 10.1007/s00253-017-8528-6. Epub 2017 Oct 2.

Abstract

Since the 1950s, Staphylococcus carnosus is used as a starter culture for sausage fermentation where it contributes to food safety, flavor, and a controlled fermentation process. The long experience with S. carnosus has shown that it is a harmless and "food grade" species. This was confirmed by the genome sequence of S. carnosus TM300 that lacks genes involved in pathogenicity. Since the development of a cloning system in TM300, numerous genes have been cloned, expressed, and characterized and in particular, virulence genes that could be functionally validated in this non-pathogenic strain. A secretion system was developed for production and secretion of industrially important proteins and later modified to also enable display of heterologous proteins on the surface. The display system has been employed for various purposes, such as development of live bacterial delivery vehicles as well as microbial biocatalysts or bioadsorbents for potential environmental or biosensor applications. Recently, this surface display system has been utilized for display of peptide and protein libraries for profiling of protease substrates and for generation of various affinity proteins, e.g., Affibody molecules and scFv antibodies. In addition, by display of fragmented antigen-encoding genes, the surface expression system has been successfully used for epitope mapping of antibodies. Reviews on specific applications of S. carnosus have been published earlier, but here we provide a more extensive overview, covering a broad range of areas from food fermentation to sophisticated methods for protein-based drug discovery, which are all based on S. carnosus.

摘要

自 20 世纪 50 年代以来,肉葡萄球菌被用作香肠发酵的起始培养物,它有助于食品安全、风味和控制发酵过程。肉葡萄球菌的长期应用经验表明,它是一种无害的“食品级”物种。这一点通过肉葡萄球菌 TM300 的基因组序列得到了证实,该序列缺乏与致病性相关的基因。自从 TM300 开发出克隆系统以来,已经克隆、表达和表征了许多基因,特别是可以在这种非致病性菌株中进行功能验证的毒力基因。开发了一种分泌系统用于生产和分泌工业上重要的蛋白质,后来对其进行了修改,以便也能够在表面展示异源蛋白质。该展示系统已用于各种目的,例如开发作为活细菌递送载体的生物以及用于潜在环境或生物传感器应用的微生物生物催化剂或生物吸附剂。最近,该表面展示系统已用于展示肽和蛋白质文库,以分析蛋白酶底物,并生成各种亲和蛋白,例如,亲和体分子和 scFv 抗体。此外,通过展示抗原编码基因的片段,表面表达系统已成功用于抗体表位作图。以前已经发表了关于肉葡萄球菌特定应用的评论,但在这里我们提供了更广泛的概述,涵盖了从食品发酵到基于蛋白质的药物发现等广泛领域,这些都基于肉葡萄球菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4722/5694512/2de13795013a/253_2017_8528_Fig1_HTML.jpg

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