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酵母中的武装技术——全细胞生物催化剂和蛋白质工程的新策略。

Arming Technology in Yeast-Novel Strategy for Whole-cell Biocatalyst and Protein Engineering.

机构信息

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.

出版信息

Biomolecules. 2013 Sep 9;3(3):632-50. doi: 10.3390/biom3030632.

Abstract

Cell surface display of proteins/peptides, in contrast to the conventional intracellular expression, has many attractive features. This arming technology is especially effective when yeasts are used as a host, because eukaryotic modifications that are often required for functional use can be added to the surface-displayed proteins/peptides. A part of various cell wall or plasma membrane proteins can be genetically fused to the proteins/peptides of interest to be displayed. This technology, leading to the generation of so-called "arming technology", can be employed for basic and applied research purposes. In this article, we describe various strategies for the construction of arming yeasts, and outline the diverse applications of this technology to industrial processes such as biofuel and chemical productions, pollutant removal, and health-related processes, including oral vaccines. In addition, arming technology is suitable for protein engineering and directed evolution through high-throughput screening that is made possible by the feature that proteins/peptides displayed on cell surface can be directly analyzed using intact cells without concentration and purification. Actually, novel proteins/peptides with improved or developed functions have been created, and development of diagnostic/therapeutic antibodies are likely to benefit from this powerful approach.

摘要

与传统的细胞内表达相比,蛋白质/肽的细胞表面展示具有许多吸引人的特点。当酵母被用作宿主时,这种武装技术特别有效,因为通常需要用于功能使用的真核修饰可以添加到表面展示的蛋白质/肽中。各种细胞壁或质膜蛋白的一部分可以与感兴趣的蛋白质/肽进行基因融合以进行展示。这项技术导致了所谓的“武装技术”的产生,可以用于基础和应用研究目的。在本文中,我们描述了构建武装酵母的各种策略,并概述了这项技术在生物燃料和化学品生产、污染物去除以及与健康相关的过程(包括口服疫苗)等工业过程中的多种应用。此外,武装技术通过高通量筛选适用于蛋白质工程和定向进化,这得益于表面展示的蛋白质/肽的特征,可以使用完整细胞直接进行分析,而无需浓缩和纯化。实际上,已经创造了具有改进或开发功能的新型蛋白质/肽,并且该强大方法可能有助于开发诊断/治疗性抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6684/4030959/466814b19686/biomolecules-03-00632-g001.jpg

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