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首页> 外文期刊>Journal of Agricultural and Food Chemistry >Engineered Arabidopsis Blue Light Receptor LOV Domain Variants with Improved Quantum Yield, Brightness, and Thermostability
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Engineered Arabidopsis Blue Light Receptor LOV Domain Variants with Improved Quantum Yield, Brightness, and Thermostability

机译：工程拟南芥蓝光受体Lov结构域变体，具有改善量子产量，亮度和热稳定性

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摘要

Despite remarkable contribution of green fluorescent protein and its variants for better understanding of various biological functions, its application for anaerobic microorganisms has been limited because molecular oxygen is essential for chromophore formation. To overcome the limitation, we engineered a plant-derived light, oxygen, or voltage (LOV) domain containing flavin mononucleotide for enhanced spectral properties. The resulting LOV variants exhibited improved fluorescence intensity (20 and 70% higher for SH3 and 70% for BR1, respectively) compared to iLOV, an LOV variant isolated in a previous study, and the quantum yields of the LOV variants (0.40 for SH3 and 0.45 for BR1) were also improved relative to that of iLOV (Q = 0.37). In addition to fluorescence intensity, the identified mutations of SH3 enabled an improved thermostability of the protein. The engineered LOV variants with enhanced spectral properties could provide a valuable tool for fluorescent molecular probes under anaerobic conditions.
机译：尽管绿色荧光蛋白及其变体贡献显着贡献，但更好地了解各种生物学功能，其对厌氧微生物的应用受到限制，因为分子氧对发色团形成是必不可少的。为了克服限制，我们设计了含有黄蛋白单核苷酸的植物衍生的光，氧气或电压（LOV）结构域，用于增强光谱性能。与ILOV相比，所得LOV变体分别表现出改善的荧光强度（SH3和SH3和70％，BR1的70％），并在先前的研究中分离的LOV变异，以及LOV变异的量子产量（SH3和SH3的0.40对于BR1的0.45），相对于ILOV的0.45也得到改善（Q = 0.37）。除了荧光强度，SH3的鉴定突变使得蛋白质的热稳定性提高。具有增强的光谱性能的工程化LOV变体可以在厌氧条件下提供用于荧光分子探针的有价值的工具。

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