Development of a new yeast surface display system based on Spi1 as an anchor protein

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• 作者：Cecilia Andreu ; Marcel·lí del Olmo
• 关键词：Saccharomyces cerevisiae ; Spi1 ; Stationary phase ; Stress conditions ; Yeast surface display
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：101
• 期：1
• 页码：287-299
• 全文大小：
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Microbiology; Microbial Genetics and Genomics; Biotechnology;
• 出版者：Springer Berlin Heidelberg
• ISSN：1432-0614
• 卷排序：101

文摘

Yeast surface display is a powerful tool widely used for many biotechnological and biomedical applications. It consists in exposing peptides and proteins of interest on the surface of Saccharomyces cerevisiae and other yeasts. These molecules are fused to the amino or carboxy terminus of an appropriate cell wall protein, usually bound by glycosylphosphatidylinositol. Several systems for this purpose have been reported to date. In this work, we describe a new yeast surface display strategy based on cell wall protein Spi1 as an anchor, which is expressed in centromeric and episomal plasmids under the control of its own promoter or that corresponding to the PGK1 glycolytic gene. Exposure efficiency was demonstrated by western blot, flow cytometry analyses, and fluorescence microscopy by taking advantage of including the V5 epitope. We also demonstrated the ability of this new strategy for the exposure of several peptides and proteins of different sizes. The regulation of the SPI1 promoter by the stationary phase and other stress conditions reveals interesting potential applications of systems based on it for industrial and biotechnological processes.