Harnessing synthetic biology to enhance heterologous protein expression
Harnessing synthetic biology to enhance heterologous protein expression
复制标题
利用合成生物学增强异源蛋白表达
DOI：
10.1002/pro.3907
复制
发表时间：
2020
期刊：
影响因子：
8
通讯作者：
Hecht, Michael
中科院分区：
文献类型：
--
作者：
Zarzhitsky, Shlomo;Jiang, Alex;Stanley, Elizabeth;Hecht, Michael
在微生物宿主中表达异源蛋白的能力对于许多研究和技术领域至关重要。然而，在大多数情况下，所需蛋白的成功表达和纯化需要与另一种蛋白融合。迄今为止，所有的融合伴侣都是从天然序列中选择的，这些天然序列是为了其他目的而进化的，可能不是最佳的融合伴侣。然而，合成生物学和蛋白质设计的兴起使得利用自然界中没有的新序列来设计和优化融合蛋白成为可能。在这里，我们描述了一系列的从头表达增强蛋白（DEEP），促进高水平表达和容易纯化的异源蛋白和肽。为了测试DEEP系统，将从头蛋白融合到涵盖一系列大小和溶解度的几种靶蛋白。在所有情况下，与DEEP的融合优于与SUMO的融合，SUMO是一种常用的天然融合伴侣。可被工程化用于特定融合应用的新型蛋白质的可用性可有益于增强多种异源蛋白质的表达。
The ability to express heterologous proteins in microbial hosts is crucial for many areas of research and technology. In most cases, however, successful expression and purification of the desired protein require fusion to another protein. To date, all fusion partners have been chosen from natural sequences, which evolved for other purposes, and may not be optimal fusion partners. However, the rise of synthetic biology and protein design make it possible to design and optimize fusion proteins usingnovelsequences that did not arise in nature. Here, we describe a series of De novo Expression Enhancer Proteins (DEEPs) that facilitate high‐level expression and facile purification of heterologous proteins and peptides. To test the DEEP system, a de novo protein was fused to several target proteins covering a range of sizes and solubilities. In all cases, fusions to DEEP outperformed fusions to SUMO, a commonly used natural fusion partner. The availability of novel proteins that can be engineered for specific fusion applications could be beneficial to enhance the expression of a wide range of heterologous proteins.
影响因子：
8
作者：
Go, Abigail;Kim, Seho;Hecht, Michael H.
通讯作者：
Hecht, Michael H.