Construction of a mApple-D6A3-mediated biosensor for detection of heavy metal ions

文献类型： 外文期刊

第一作者： Ji, Yangyang

作者： Ji, Yangyang;Zhou, Xin;Liu, Daling;Ji, Yangyang;Guan, Feifei;Zhou, Xin;Liu, Xiaoqing;Wu, Ningfeng;Tian, Jian

作者机构：

关键词： Biosensor; Heavy metal; Detection

期刊名称：AMB EXPRESS （ 影响因子：3.298； 五年影响因子：3.427 ）

ISSN： 2191-0855

年卷期： 2020 年 10 卷 1 期

页码：

收录情况： SCI

摘要： Pollution of heavy metals in agricultural environments is a growing problem to the health of the world's human population. Green, low-cost, and efficient detection methods can help control such pollution. In this study, a protein biosensor, mApple-D6A3, was built from rice-derived Cd2+-binding protein D6A3 fused with the red fluorescent protein mApple at the N-terminus to detect the contents of heavy metals. Fluorescence intensity of mApple fused with D6A3 indicated the biosensor's sensitivity to metal ions and its intensity was more stable under alkaline conditions. mApple-D6A3 was most sensitive to Cu2+, then Ni2+, then Cd2+. Isothermal titration calorimetry experiments demonstrated that mApple-D6A3 successfully bound to each of these three metal ions, and its ability to bind the ions was, from strongest to weakest, Cu2+ > Cd2+ > Ni2+. There were strong linear relationships between the fluorescence intensity of mApple-D6A3 and concentrations of Cd2+ (0-100 mu M), Cu2+ (0-60 mu M) and Ni2+ (0-120 mu M), and their respective R-2 values were 0.994, 0.973 and 0.973. When mApple-D6A3 was applied to detect concentrations of heavy metal ions in water (0-0.1 mM) or culture medium (0-1 mM), its accuracy for detection attained more than 80%. This study demonstrates the potential of this biosensor as a tool for detection of heavy metal ions.

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