Translation related factors improve the productivity of a Streptomyces-based cell-free protein synthesis system
Translation related factors improve the productivity of a Streptomyces-based cell-free protein synthesis system
复制标题
翻译相关因素提高了基于链霉菌的无细胞蛋白质合成系统的生产力
DOI：
10.1021/acssynbio.0c00140
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发表时间：
2020
影响因子：
4.7
通讯作者：
Li Jian
中科院分区：
文献类型：
--
作者：
Xu Huiling;Liu Wan-Qiu;Li Jian
无细胞蛋白质合成(CFPS)系统正在成为体外生产重组蛋白质的有效平台。为了能够表达各种蛋白质，不同的CFPS系统已经被开发出来，以更好地模拟本地宿主的细胞环境。在这种背景下，最近开发了一个基于链霉菌的CFPS系统来表达高GC含量的基因。遗憾的是，初始体系的蛋白质产量相对较低(∼为50μg/mL)。在这里，我们试图解决这一限制，并提高基于链霉菌的CFPS系统的生产率。通过在CFPS反应中加入蛋白质翻译相关因子，我们能够获得大约400μg/mL的蛋白质产量，这是迄今为止报道的最高产量。我们期待我们的增强型链霉菌CFPS系统将为代谢工程和合成生物学中的新应用奠定基础，例如在天然产物的体外发现和合成方面，这些天然产物是由具有高GC含量(>；70%)基因组的链霉菌产生的。
Cell-free protein synthesis (CFPS) systems are emerging as effective platforms for the production of recombinant proteinsin vitro. To enable the expression of various proteins, different CFPS systems have been developed to better mimic the cellular environment of native hosts. In this context, aStreptomyces-based CFPS system was recently developed to express high GC-content genes. Unfortunately, protein yields from the initial system were relatively low (∼50 μg/mL). Here, we sought to address this limitation and enhance the productivity of theStreptomyces-based CFPS system. By adding protein translation related factors to CFPS reactions, we were able to achieve protein yields of approximately 400 μg/mL, which is the highest yield reported to date. We expect our enhancedStreptomycesCFPS system will set the stage for novel applications in metabolic engineering and synthetic biology such asin vitrodiscovery and synthesis of natural products, which are produced byStreptomycesspecies with high GC-content (>70%) genomes.