The novel hydrolase IpcH initiates the degradation of isoprocarb in a newly isolated strain Rhodococcus sp. D-6

Isoprocarb (IPC), a representative monocyclic carbamate insecticide, poses risks of environmental contamination and harm to non-target organisms. However, its degradation mechanism has not been reported. In this study, a newly IPC-degrading strain D-6 was isolated from the genus Rhodococcus, and its degradation characteristics and pathway of IPC were analyzed. A novel hydrolase IpcH, responsible for hydrolyzing IPC to 2-isopropylphenol (IPP), was identified. IpcH exhibited low similarity (< 27%) with other reported hydrolases, including previously characterized carbamate insecticides hydrolases, indicating its novelty. The Km and kcat values of IpcH towards IPC were 69.99 ± 8.33 μM and 95.96 ± 4.02 s-1, respectively. Also, IpcH exhibited catalytic activity towards various types of carbamate insecticides, including monocyclic carbamates (IPC, fenobucarb and propoxur), bicyclic carbamates (carbaryl and carbofuran), and linear carbamates (oxamyl and aldicarb). The molecular docking and site-directed mutagenesis revealed that His254, His256, His329 and His376 were essential for IpcH activity. Strain D-6 can effectively reduce the toxicity of IPC and IPP towards sensitive organisms through its degradation ability. This study presents the initial report on IPC degradation pathway and molecular mechanism of IPC degradation, and provides a good potential strain for bioremediating IPC and IPP-contaminated environments. IPC exerts insecticidal function by inhibiting activity of AchE which is also present in human and other non-target organisms. Consequently, IPC causes toxicity to them and poses risks to ecosystem. IPC is classed as moderately hazardous in the World Health Organization (WHO) Acute Hazard Ranking. Therefore, it is imperative to investigate IPC degradation mechanism and environmental behavior. Microbial degradation is a promising method for remediating polluted environments. Here, an IPC-degrading strain Rhodococcus sp. D-6 was isolated. The degradation pathway of IPC and its molecular mechanism were studied, and a potential strain for bioremediation of IPC and IPP-contaminated environments was provided.