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首页> 外文期刊>Applied Microbiology and Biotechnology >Comparative analysis of twin-arginine (Tat)-dependent protein secretion of a heterologous model protein (GFP) in three different Gram-positive bacteria
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Comparative analysis of twin-arginine (Tat)-dependent protein secretion of a heterologous model protein (GFP) in three different Gram-positive bacteria

机译：在三种不同的革兰氏阳性细菌中异源模型蛋白（GFP）的双精氨酸（Tat）依赖性蛋白分泌的比较分析

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摘要

In contrast to the general protein secretion (Sec) system, the twin-arginine translocation (Tat) export pathway allows the translocation of proteins across the bacterial plasma membrane in a fully folded conformation. Due to this feature, the Tat pathway provides an attractive alternative to the secretory production of heterologous proteins via the Sec system. In this study, the potential for Tat-dependent heterologous protein secretion was compared in the three Gram-positive bacteria Staphylococcus carnosus, Bacillus subtilis, and Corynebacterium glutamicum using green fluorescent protein (GFP) as a model protein. In all three microorganisms, fusion of a Tat signal peptide to GFP resulted in its Tat-dependent translocation across the corresponding cytoplasmic membranes. However, striking differences with respect to the final localization and folding status of the exported GFP were observed. In S. carnosus, GFP was trapped entirely in the cell wall and not released into the supernatant. In B. subtilis, GFP was secreted into the supernatant, however, in an inactive form. In contrast, C. glutamicum effectively secreted active GFP. Our results clearly demonstrate that a comparative evaluation of different Gram-positive host microorganisms is a crucial step on the way to an efficient Tat-mediated secretory production process for a desired heterologous target protein.
机译：与一般的蛋白质分泌（Sec）系统相比，双精氨酸易位（Tat）出口途径允许蛋白以完全折叠的构型跨细菌质膜移位。由于此功能，Tat途径为通过Sec系统分泌异源蛋白质提供了有吸引力的替代方法。在这项研究中，使用绿色荧光蛋白（GFP）作为模型蛋白，比较了三种革兰氏阳性细菌肉食葡萄球菌，枯草芽孢杆菌和谷氨酸棒杆菌中Tat依赖的异源蛋白分泌的潜力。在所有三种微生物中，Tat信号肽与GFP融合均导致其Tat依赖性转位穿过相应的细胞质膜。但是，观察到有关输出的GFP的最终定位和折叠状态的显着差异。在肉食链球菌中，GFP完全被捕获在细胞壁中，而不释放到上清液中。在枯草芽孢杆菌中，GFP以无活性形式分泌到上清液中。相反，谷氨酸棒杆菌有效地分泌了活性GFP。我们的结果清楚地表明，对不同的革兰氏阳性宿主微生物进行比较评估是有效的Tat介导的所需异源靶蛋白分泌过程的关键步骤。

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