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 using novel sequences 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.

中文翻译：

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利用合成生物学增强异源蛋白表达。

在微生物宿主中表达异源蛋白质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，所需蛋白质的成功表达和纯化需要与另一种蛋白质融合。迄今为止，所有融合伴侣都是从自然序列中选择的，这些序列是为了其他目的而进化的，并且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使用自然界中不存在的新序列设计和优化融合蛋白成为可能。在这里，我们描述了一系列从头表达增强蛋白 (DEEP)，它们有助于异源蛋白和肽的高水平表达和轻松纯化。为了测试 DEEP 系统，将一种从头蛋白质与多种大小和溶解度范围内的目标蛋白质融合。在所有情况下，与 DEEP 的融合优于与常用的自然融合伙伴 SUMO 的融合。可针对特定融合应用进行工程设计的新型蛋白质的可用性可能有利于增强多种异源蛋白质的表达。