Enhanced biodegradation potential of Klebsiella michiganensis ES15 for acetochlor: Gene knockout, heterologous expression, molecular docking, and bioremediation

文献类型： 外文期刊

第一作者： Zhao, Minghui

作者： Zhao, Minghui;Xiao, Yufeng;Yang, Bingbing;Wang, Siya;Tian, Bowen;Zhang, Hao;Wu, Xian

作者机构：

关键词： Biodegrdation; Molecular docking; Functional gene; qPCR; Klebsiella michiganensis ES15; Acetochlor

期刊名称：PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY （ 影响因子：4.0； 五年影响因子：4.5 ）

ISSN： 0048-3575

年卷期： 2025 年 214 卷

页码：

收录情况： SCI

摘要： Long-term large-scale application of acetochlor has led to its accumulation in soil, causing serious environmental pollution. In this study, Klebsiella michiganensis ES15 was isolated from the contaminated reactive sludge of an acetochlor pesticide plant and achieved 79.23 % degradation of acetochlor within 4 d after medium optimization using response surface methodology. The optimal degradation conditions for strain ES15 were pH 7, 20 mg L-1 acetochlor, and 3 % inoculum size with tolerance to Fe3+ and Zn2+. Strain ES15 contained functional genes, including gst, HylA, amoA, and nahA, which were related to acetochlor degradation, with gst upregulated 16.28-fold, indicating that it was a major functional gene. And the Km-gst gene was identified as a key gene in acetochlor biodegradation, as its knockout resulted in an approximately 40 % reduction in degradation efficiency. Strain ES15 catalyzed the breakdown of acetochlor into 2-methyl-6-ethyl-2-chloroacetanilide, which was subsequently hydrolyzed into 2-methyl-6-ethylaniline. Oxidative stress enzyme activity initially increased and then decreased during degradation. Molecular docking was performed between gst and acetochlor. Additionally, qPCR analysis demonstrated the upregulation of antioxidant-related genes (sodB, katA, tpx, and gshA) and biofilmrelated genes (luxS, rhII, lasR, and waaE), with sodB and waaE upregulated by 14.36-and 18.84-fold, respectively. Strain ES15 degraded 79.9 % of acetochlor in unsterilized soil over 28 d and demonstrated good colonization ability. Furthermore, strain ES15 mitigated acetochlor-induced damage to spinach that is a sensitive crop. This study provides a promising strategy for effective degradation of acetochlor.

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