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选定真菌菌株产生羧酸酯酶用于生物降解和利用联苯菊酯杀虫剂作为碳源的能力:体外和计算机模拟方法。

The ability of selected fungal strains to produce carboxylesterase enzymes for biodegradation and use of bifenthrin insecticide as carbon source: in vitro and in silico approaches.

机构信息

Department of Biotechnology, COMSATS University Islamabad, Abbottabad Campus, Abbottabad, 22060, Pakistan.

Department of Environmental Sciences, COMSATS University Islamabad, Abbottabad Campus, Abbottabad, 22060, Pakistan.

出版信息

Bioprocess Biosyst Eng. 2024 Oct;47(10):1691-1705. doi: 10.1007/s00449-024-03062-9. Epub 2024 Jul 19.

Abstract

Bifenthrin (BF) is a broad-spectrum type I pyrethroid insecticide that acts on insects by impairing the nervous system and inhibiting ATPase activity, and it has toxic effects on non-target organisms and high persistence in the environment. This study aimed to determine the potential of six different fungi, including Pseudozyma hubeiensis PA, Trichoderma reesei PF, Trichoderma koningiopsis PD, Purpureocillium lilacinum ACE3, Talaromyces pinophilus ACE4, and Aspergillus niger AJ-F3, to degrade BF. Three different concentrations of BF, including 0.1%, 0.2%, and 0.3% w/v, were used in the sensitivity testing that revealed a significant (p ≤ 0.01) impact of BF on fungal growth. Enzymatic assays demonstrated that both intracellular and extracellular carboxylesterases hydrolyzed BF with the enzymatic activity of up to 175 ± 3 U (μmol/min) and 45 ± 1 U, respectively. All tested fungi were capable of utilizing BF as a sole carbon source producing 0.06 ± 0.01 to 0.45 ± 0.01 mg dry biomass per mg BF. Moreover, the presence of PytH was determined in the fungi using bioinformatics tools and was found in A. niger, T. pinophilus, T. reesei, and P. lilacinum. 3D structures of the PytH homologs were predicted using AlphaFold2, and their intermolecular interactions with pyrethroids were determined using MOE. All the homologs interacted with different pyrethroids with a binding energy of lesser than - 10 kcal/mol. Based on the study, it was concluded that the investigated fungi have a greater potential for the biodegradation of BF.

摘要

溴氰菊酯(BF)是一种广谱 I 型拟除虫菊酯杀虫剂,通过损害神经系统和抑制 ATP 酶活性来作用于昆虫,对非靶标生物有毒性,在环境中持久性高。本研究旨在确定六种不同真菌(包括湖北假丝酵母 PA、里氏木霉 PF、康宁木霉 PD、紫色拟青霉 ACE3、塔宾曲霉 ACE4 和黑曲霉 AJ-F3)对 BF 的降解潜力。在灵敏度测试中使用了三种不同浓度的 BF,包括 0.1%、0.2%和 0.3%w/v,结果表明 BF 对真菌生长有显著影响(p≤0.01)。酶活性测定表明,胞内和胞外羧酸酯酶均能水解 BF,酶活性分别高达 175±3 U(μmol/min)和 45±1 U。所有测试真菌均能利用 BF 作为唯一碳源,产生 0.06±0.01 至 0.45±0.01 mg 干生物量/mg BF。此外,还使用生物信息学工具在真菌中检测到了 PytH 的存在,在黑曲霉、塔宾曲霉、里氏木霉和紫色拟青霉中均发现了 PytH。使用 AlphaFold2 预测了 PytH 同源物的 3D 结构,并使用 MOE 确定了它们与拟除虫菊酯的分子间相互作用。所有同源物都与不同的拟除虫菊酯相互作用,结合能小于-10 kcal/mol。基于研究结果,可以得出结论,所研究的真菌对 BF 的生物降解具有更大的潜力。

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