Identification of fungal enzymes involving 3-phenoxybenzoic acid degradation by using enzymes inhibitors and inducers

Jiayuan Zhao,Xiaofeng Chen,Dongying Jia,Kai Yao

doi:10.1016/j.mex.2019.100772

Jiayuan Zhao, Xiaofeng Chen + Show 2 more

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https://doi.org/10.1016/j.mex.2019.100772

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Journal: MethodsX	Publication Date: Dec 17, 2019
Citations: 3	License type: cc-by

Affiliation: Sichuan Normal University, Sichuan University

Abstract

Pyrethroid residues in food and the environment can be bio-transformed into 3-phenoxybenzoic acid (3-PBA); It is more toxic than the parent compounds, and has been detected in milk, soil, and human urine. In this study, when incubated at 30 °C and 180 rpm for 48 h, mycelial pellets during logarithmic growth phase were obtained and washed 2 times by phosphate buffer. The cell debris solutions and filter liquor from inducible and non-inducible samples were cultured with 3-PBA and its intermediate metabolites at same condition, and the location and induction of enzymes were analyzed by the degradation. Then Cytochrome P450 (CYP450), lignin peroxidase (LiP), laccase, manganese peroxidase (MnP), and dioxygenase were selected as candidate enzymes due to these oxidases existing in the fungi and capable of degrading the contaminants with similar structures of these compounds, and CuSO4, NaN3, AgNO3, EDTA or piperonyl butoxide (PBO) were used as the enzymes inhibitors and inducers. The degradation of 3-PBA and its intermediate metabolites and the fungal biomass in presence of enzymes inhibitors and inducers was arranged to analyze the possible degrading-enzymes, and the co-metabolic enzymes and pathways can be reasoned. This study provided a promising method for studying the co-metabolic enzymes of 3-PBA degradation by fungi.•The presented MethodsX was conducted for co-metabolic enzymes and pathways of 3-PBA degradation.•The culturing condition for presenting enzyme properties were investigated.•The candidate enzymes were analyzing based on location, induction of enzymes, fungal enzyme systems and chemical structures of these compounds.

Highlights

Pyrethroid residues in food and the environment can be bio-transformed into 3-phenoxybenzoic acid (3-PBA); It is more toxic than the parent compounds, and has been detected in milk, soil, and human urine
The inhibitors and inducers of CYP 450, lignin peroxidase (LiP), laccase, manganese peroxidase (MnP), and dioxygenase were selected based on the results reported by Ikehata et al (2004) [7], Oszajca et al (2016) [8], Sari et al (2012) [6], Senthivelan et al (2016) [9] and Yeom and Sligar (1997) [10]
As an intermediate metabolite of pyrethroids, 3-phenoxybenzoic acid (3-PBA) is more toxic than its parent compounds and has been detected in milk, soil, and human urine [3,4,5]. 3-PBA can be metabolized to less potent metabolites through microbial degradation

Summary

Introduction

Pyrethroid residues in food and the environment can be bio-transformed into 3-phenoxybenzoic acid (3-PBA); It is more toxic than the parent compounds, and has been detected in milk, soil, and human urine. The cell debris solutions and filter liquor from inducible and noninducible samples were cultured with 3-PBA and its intermediate metabolites at same condition, and the location and induction of enzymes were analyzed by the degradation. The degradation of 3-PBA and its intermediate metabolites and the fungal biomass in presence of enzymes inhibitors and inducers was arranged to analyze the possible degrading-enzymes, and the co-metabolic enzymes and pathways can be reasoned. This study provided a promising method for studying the co-metabolic enzymes of 3-PBA degradation by fungi

Methods

Findings

Conclusion