Genomic, Transcriptomic and Suspect/Non-Target Screening Analyses Reveal the Role of CYP450s in the Degradation of Imazalil and Delineate Its Transformation Pathway by Cladosporium herbarum

IF 5.7 2区 生物学
Christina V. Papazlatani, Sotirios Vasileiadis, Eleni I. Panagopoulou, Dimitrios E. Damalas, Panagiotis A. Karas, Evdoxia Gerovasileiou, Nikolaos S. Thomaidis, Dimitrios G. Karpouzas
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Abstract

Imazalil (IMZ), a major surface water contaminant characterised by high environmental recalcitrance and toxicity, is used in fruit-packaging plants to control fungal infestations during storage. This leads to the production of wastewaters which should be treated on site before their environmental release. We previously isolated a Cladosporium herbarum strain, the first microorganism that could degrade IMZ. Here we describe the genetic network utilised by the fungus to degrade IMZ and its detailed transformation. Genomic and transcriptomic analysis of C. herbarum pointed to the involvement of strongly upregulated CYP450s in IMZ degradation, as further verified by cessation of its biodegradation by CYP450 inhibitors. LC-QTOF-HRMS analysis and suspect/non-target screening identified nine transformation products (TPs) of IMZ. IMZ biotransformation mainly proceeded through O-dealkylation, while other less important paths, most probably controlled by upregulated oxidases, were operative involving successive hydroxylation reactions. These lead to the formation of TPs like IMZ_313 and IMZ_331, with the former being further transformed through imidazole ring scission to IMZ_288, a TP reported for the first time. We provide first evidence for the transformation mechanism of IMZ by C. herbarum and the potential genes/enzymes involved, paving the way for the use of C. herbarum in the biodepuration of agro-industrial effluents.

Abstract Image

基因组学、转录组学和可疑/非靶标筛选分析揭示了cyp450在植物枝孢菌(Cladosporium herbarum)降解Imazalil中的作用,并描绘了其转化途径
Imazalil (IMZ)是一种主要的地表水污染物,具有高度的环境抗性和毒性,用于水果包装厂在储存期间控制真菌感染。这导致了废水的产生,这些废水在排放到环境之前应该在现场进行处理。我们之前分离了一株草本枝孢菌,这是第一个可以降解IMZ的微生物。在这里,我们描述了真菌利用遗传网络来降解IMZ及其详细的转化。对C. herbarum的基因组学和转录组学分析表明,CYP450的强烈上调参与了IMZ的降解,CYP450抑制剂停止了IMZ的生物降解,这进一步证实了这一点。LC-QTOF-HRMS分析和可疑/非目标筛选鉴定出9个IMZ转化产物。IMZ的生物转化主要通过o -脱烷基进行,而其他不太重要的途径,很可能由上调的氧化酶控制,涉及连续的羟基化反应。这导致形成像IMZ_313和IMZ_331这样的TP,其中IMZ_331通过咪唑环裂解进一步转化为IMZ_288,这是首次报道的TP。本研究首次揭示了香茅对IMZ的转化机制和可能涉及的基因/酶,为香茅在农工废水生物净化中的应用铺平了道路。
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来源期刊
Microbial Biotechnology
Microbial Biotechnology Immunology and Microbiology-Applied Microbiology and Biotechnology
CiteScore
11.20
自引率
3.50%
发文量
162
审稿时长
1 months
期刊介绍: Microbial Biotechnology publishes papers of original research reporting significant advances in any aspect of microbial applications, including, but not limited to biotechnologies related to: Green chemistry; Primary metabolites; Food, beverages and supplements; Secondary metabolites and natural products; Pharmaceuticals; Diagnostics; Agriculture; Bioenergy; Biomining, including oil recovery and processing; Bioremediation; Biopolymers, biomaterials; Bionanotechnology; Biosurfactants and bioemulsifiers; Compatible solutes and bioprotectants; Biosensors, monitoring systems, quantitative microbial risk assessment; Technology development; Protein engineering; Functional genomics; Metabolic engineering; Metabolic design; Systems analysis, modelling; Process engineering; Biologically-based analytical methods; Microbially-based strategies in public health; Microbially-based strategies to influence global processes
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