期刊
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
卷 9, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fbioe.2021.799781
关键词
cadmium detection; negative feedback amplifier; sensitivity; specificity; whole-cell biosensor
资金
- National Key Research and Development Program of China [2018YFA0902100]
- National Natural Science Foundation of China [22178262, 21576197]
A Cd2+ whole-cell biosensor with a negative feedback amplifier exhibited significantly improved sensitivity and specificity compared to traditional biosensors. This biosensor showed a lower detection limit and increased tolerance to Cd2+, making it suitable for real field applications. The negative feedback amplifier also provided faster response time and higher specificity for Cd2+ detection, showing its potential for enhancing biosensor performance.
Although many whole-cell biosensors (WCBs) for the detection of Cd2+ have been developed over the years, most lack sensitivity and specificity. In this paper, we developed a Cd2+ WCB with a negative feedback amplifier in P. putida KT2440. Based on the slope of the linear detection curve as a measure of sensitivity, WCB with negative feedback amplifier greatly increased the output signal of the reporter mCherry, resulting in 33% greater sensitivity than in an equivalent WCB without the negative feedback circuit. Moreover, WCB with negative feedback amplifier exhibited increased Cd2+ tolerance and a lower detection limit of 0.1 nM, a remarkable 400-fold improvement compared to the WCB without the negative feedback circuit, which is significantly below the World Health Organization standard of 27 nM (0.003 mg/L) for cadmium in drinking water. Due to the superior amplification of the output signal, WCB with negative feedback amplifier can provide a detectable signal in a much shorter time, and a fast response is highly preferable for real field applications. In addition, the WCB with negative feedback amplifier showed an unusually high specificity for Cd2+ compared to other metal ions, giving signals with other metals that were between 17.6 and 41.4 times weaker than with Cd2+. In summary, the negative feedback amplifier WCB designed in this work meets the requirements of Cd2+ detection with very high sensitivity and specificity, which also demonstrates that genetic negative feedback amplifiers are excellent tools for improving the performance of WCBs.
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