Evolved Biosensor with High Sensitivity and Specificity for Measuring Cadmium in Actual Environmental Samples

生物传感器 检出限 化学 细菌 荧光 污染 环境化学 色谱法 生物 生物化学 生态学 遗传学 量子力学 物理 有机化学
作者
Yeshen Cai,Kaili Zhu,Liang Shen,Jie Ma,Lingzhi Bao,Dongdong Chen,Liangchen Wei,Nan Wei,Binmei Liu,Yuejin Wu,Shaopeng Chen
出处
期刊:Environmental Science & Technology [American Chemical Society]
卷期号:56 (14): 10062-10071 被引量:21
标识
DOI:10.1021/acs.est.2c00627
摘要

Bacterial biosensors have great potential in contaminant detection for sensitivity, specificity, cost-effectiveness, and easy operation. However, the existing cadmium-responsive bacterial biosensors cannot meet the real-world detection requirements due to lack of sensitivity, specificity, and anti-interference capability. This study aimed to develop a bacterial biosensor for detecting the total and extractable cadmium in actual environmental samples. We constructed the cadmium-responsive biosensor with the regulatory element (cadmium resistance transcriptional regulatory, CadR) and the reporting element (GFP) and improved its performance by directed evolution. The mutant libraries of biosensors were generated by error-prone PCR and screened by continuous five-round fluorescence-activated cell sorting (FACS), and a bacteria variant epCadR5 with higher performance was finally isolated. Biosensor fluorescence intensity was measured by a microplate reader, and results showed that the evolved cadmium-responsive bacterial biosensor was of high sensitivity and specificity in detecting trace cadmium, with a detection limit of 0.45 μg/L, which is 6.8 times more specific to cadmium than that of the wild-type. Furthermore, microscopic qualitative analysis results showed that the bacteria could produce fluorescence response in a cadmium-contaminated soil matrix, and quantitative analysis results showed that the values of cadmium from epCadR5 bacteria were close to that from inductively coupled plasma-mass spectrometry. These results suggest that the biosensor may have a broad application prospect in the detection of cadmium-contaminated soil and water.
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