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揭示了K+对大肠杆菌中xanthorhodopsin表达的关键作用

Chenda Hour¹, Kimleng Chuon¹, Myung-Chul Song¹

¹Department of Life Science and Institute of Biological Interfaces, Sogang University, Seoul, South Korea.

Journal of photochemistry and photobiology. B, Biology

|July 13, 2024

概括

我们通过优化K+水平和诱导度,成功地在大肠杆菌中表达了xanthodopsin (XR). 突变,比如用氨酸取代Ser-159,显著增强XR表达和色素形成,改善蛋白质产量.

科学领域:

微生物学和结构生物学
蛋白质表达和生物化学

背景情况:

来自Salinibacter ruber的光驱动质子Xanthorhodopsin (XR) 使用视网膜和salinixanthin (SAL),但面临表达挑战.
之前对XR的研究受限于难以实现野生类型和突变形式的稳定,高水平表达的困难.

研究的目的:

克服表达限制并优化野生类型和突变类型的ksanthodopsin (XR) 的生产.
研究特定氨基酸残留物和K+在XR表达和色素形成中的作用.

主要方法:

在K+的存在下,工程化Escherichia coli用于野生类型和突变XR的异质表达.
优化K+度和诱导剂水平以实现高XR生产.
描述了表达的XR变体的质子和光循环动力学.

主要成果:

在E. coli中成功地实现了野生型和突变XR的高水平表达,具有优化的K +和低诱导度.
确定了Ser-159作为一个关键残留物;对氨酸的突变显著增强了XR表达,表明了硬质阻碍.
表达XR的质子和光循环特性与原生XR相似,验证了表达系统.

结论:

在大肠杆菌中,K+对于高效的XR表达至关重要,可能模仿其在性生物中的作用.
-159突变提供了一种克服硬质障碍的策略,并改善XR色素形成和整体产量.
盐的选择为提高生物技术应用中的蛋白质生产和功能提供了一个可行的方法.

关键词:

由光驱动的外向质子向外.微生物罗多普辛是一种微生物.沙利尼巴克特 (Salinibacter) 已经被了.盐酸素 (Salinixanthin) 是一种含盐量较大的物质.桑托尔多多普辛 (Xanthorhodopsin) 是一种色素.

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