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首页> 外文期刊>Biotechnology Progress >Development of Surface-Engineered Yeast Cells Displaying Phytochelatin Synthase and Their Application to Cadmium Biosensors by the Combined Use of Pyrene-Excimer Fluorescence
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Development of Surface-Engineered Yeast Cells Displaying Phytochelatin Synthase and Their Application to Cadmium Biosensors by the Combined Use of Pyrene-Excimer Fluorescence

机译：yr-准分子荧光联合使用显示植物螯合酶合酶的表面工程酵母细胞的发展及其在镉生物传感器中的应用

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The development of simple, portable, inexpensive, and rapid analytical methods for detecting and monitoring toxic heavy metals are important for the safety and security of humans and their environment. Herein, we describe the application of phytochelatin (PC) synthase, which plays a critical role in heavy metal responses in higher plants and green algae, in a novel fluorescent sensing platform for cadmium (Cd). We first created surface-engineered yeast cells on which the PC synthase from Arabidopsis (AtPCSl) was displayed with retention of enzymatic activity. The general concept for the sensor is based on the Cd level-dependent synthesis of PC2 from glutathiones by AtPCSl-displaying yeast cells, followed by simple discriminative detection of PC2 via sensing of excimer fluorescence of thiol-labeling pyrene probes. The intensity of excimer fluorescence increased in the presence of Cd up to 1.0 μM in an approximately dose-dependent manner. This novel biosensor achieved a detection limit of as low as 0.2 μM (22.5 μg/L) for Cd. Although its use may be limited by the fact that Cu and Pb can induce cross-reaction, the proposed simple biosensor holds promise as a method useful for cost-effective screening of Cd contamination in environmental and food samples. The AtPCSl -displaying yeast cells also might be attractive tools for dissection of the catalytic mechanisms of PCS.
机译：用于检测和监测有毒重金属的简单，便携式，廉价和快速的分析方法的开发对于人类及其环境的安全与保障至关重要。在这里，我们描述了植物螯合酶（PC）合酶的应用，该酶在镉（Cd）的新型荧光传感平台中在高等植物和绿藻中的重金属响应中起关键作用。我们首先创建了表面工程化的酵母细胞，其上展示了来自拟南芥属的PC合酶（AtPCS1）并具有酶促活性。传感器的一般概念是基于显示AtPCS1的酵母细胞从谷胱甘肽的Cd水平依赖性合成PC2，然后通过感测硫醇标记的pyr探针的准分子荧光对PC2进行简单的判别检测。在镉存在下，准分子荧光的强度以大约剂量依赖的方式增加至1.0μM。这种新型生物传感器对Cd的检测限低至0.2μM（22.5μg/ L）。尽管其使用可能会受到铜和铅可能引起交叉反应的事实的限制，但拟议的简单生物传感器有望作为一种经济有效地筛选环境和食品样品中镉污染的方法。展示AtPCS1的酵母细胞也可能是解剖PCS催化机制的有吸引力的工具。

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