Kuroda Kouichi, Ueda Mitsuyoshi
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
Methods Mol Biol. 2022;2491:627-641. doi: 10.1007/978-1-0716-2285-8_26.
The yeast surface display system is a valuable platform for constructing cells with novel functions for various applications and high-throughput screening of protein or peptide libraries containing random mutations. Among the host microorganisms used for surface display, yeast is the most suitable microorganism for surface engineering owing to its eukaryotic features. In yeast, proper folding and glycosylation of expressed eukaryotic proteins can be performed. Furthermore, in this system, multiple kinds of proteins can be simultaneously displayed on the cell surface. This allows for a synergistic effect between the displayed enzymes, leading to an efficient multistep reaction. Alternatively, the ratio of the enzymes to be displayed can be controlled by the co-culture of surface-engineered yeasts displaying a single kind of enzyme. Therefore, yeast surface display systems have been applied to the construction of various whole-cell biocatalysts. Here, we describe methods for the simultaneous display of multiple kinds of proteins on the yeast cell surface.
酵母表面展示系统是一个有价值的平台，可用于构建具有多种新功能的细胞，以用于各种应用以及对含有随机突变的蛋白质或肽库进行高通量筛选。在用于表面展示的宿主微生物中，酵母因其真核生物特性而成为最适合进行表面工程的微生物。在酵母中，表达的真核蛋白能够进行正确折叠和糖基化。此外，在该系统中，多种蛋白质可同时展示在细胞表面。这使得展示的酶之间产生协同效应，从而实现高效的多步反应。或者，通过共培养展示单一酶的表面工程酵母，可以控制要展示的酶的比例。因此，酵母表面展示系统已被应用于构建各种全细胞生物催化剂。在此，我们描述在酵母细胞表面同时展示多种蛋白质的方法。
