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Characterization of a novel amidohydrolase with promiscuous esterase activity from a soil metagenomic library and its application in degradation of amide herbicides

journal contribution
posted on 2024-02-12, 11:30 authored by Shengwei Sun, Wanqi Chen, Kailin Peng, Xueyingzi Chen, Jinju ChenJinju Chen

Amide herbicides have been extensively used worldwide and have received substantial attention due to their adverse environmental effects. Here, a novel amidohydrolase gene was identified from a soil metagenomic library using diethyl terephthalate (DET) as a screening substrate. The recombinant enzyme, AmiH52, was heterologously expressed in Escherichia coli and later purified and characterised, with the highest activity occurring at 40 ℃ and pH 8.0. AmiH52 was demonstrated to have both esterase and amidohydrolase activities, which exhibited highly specific activity for p-nitrophenyl butyrate (2669 U/mg) and degrading activity against several amide herbicides. In particular, it displayed the strongest activity against propanil, with a high degradation rate of 84% at 8 h. A GC-MS analysis revealed that propanil was transformed into 3,4- dichloroaniline (3,4-DCA) during this degradation. The molecular interactions and binding stability were then analysed by molecular docking and molecular dynamics simulation, which revealed that several key amino acid residues, including Tyr164, Trp66, Ala59, Val283, Arg58, His33, His191, and His226, are involved in the specific interactions with propanil. This study provides a function-driven screening method for amide herbicide hydrolase from the metagenomic libraries and a promising propanil degrading enzyme (AmiH52) for potential applications in environmental remediation.

Funding

Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX21_0581), China

History

School

  • Aeronautical, Automotive, Chemical and Materials Engineering

Department

  • Materials

Published in

Environmental Science and Pollution Research

Publisher

Springer

Version

  • AM (Accepted Manuscript)

Acceptance date

2024-02-03

ISSN

0944-1344

eISSN

1614-7499

Language

  • en

Depositor

Prof Jinju Chen. Deposit date: 11 February 2024

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