Engineering chlorpyrifos-degrading Stenotrophomonas sp. YC-1 for heavy metal accumulation and enhanced chlorpyrifos degradation

Many ecosystems are currently co-contaminated with pesticides and heavy metals, such as chlorpyrifos and cadmium. A promising strategy to remediate mixed chlorpyrifos-cadmium-contaminated sites is the use of chlorpyrifos-degrading bacteria endowed with cadmium removal capabilities. In this work, a gene coding for synthetic phytochelatins (EC20) with high cadmium-binding capacity was introduced into a chlorpyrifos-degrading bacterium, Stenotrophomonas sp. YC-1, resulting in an engineered strain with both cadmium accumulation and chlorpyrifos degradation capabilities. To improve the cadmium-binding efficiency of whole cells, EC20 was displayed on the cell surface of Stenotrophomonas sp. YC-1 using the truncated ice nucleation protein (INPNC) anchor. The surface localization of the INPNC-EC20 fusion protein was demonstrated by cell fractionation, Western blot analysis, and immunofluorescence microscopy. Expression of EC20 on the cell surface not only improved cadmium binding, but also alleviated the cellular toxicity of cadmium. As expected, the chlorpyrifos degradation rate was reduced in the presence of cadmium for cells without EC20 expression. However, expression of EC20 (higher cadmium accumulation) completely restored the level of chlorpyrifos degradation. These results demonstrated that EC20 expression not only enhanced cadmium accumulation, but also reduced the toxic effect of cadmium on chlorpyrifos degradation.