Zhao Jiayuan, Chen Xiaofeng, Jia Dongying, Yao Kai
College of Life Science, Sichuan Normal University, 610101, Chengdu, Sichuan, PR China.
College of Biomass Science and Engineering, Sichuan University, 610065, Chengdu, Sichuan, PR China.
MethodsX. 2019 Dec 17;7:100772. doi: 10.1016/j.mex.2019.100772. eCollection 2020.
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 CuSO, NaN, AgNO, 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.
食品和环境中的拟除虫菊酯残留可生物转化为3-苯氧基苯甲酸(3-PBA);它比母体化合物毒性更大,已在牛奶、土壤和人类尿液中被检测到。在本研究中,将对数生长期的菌丝球在30°C和180 rpm条件下培养48小时,然后用磷酸盐缓冲液洗涤2次。将诱导型和非诱导型样品的细胞碎片溶液和滤液在相同条件下与3-PBA及其中间代谢产物一起培养,并通过降解分析酶的定位和诱导情况。然后,由于这些氧化酶存在于真菌中且能够降解与这些化合物结构相似的污染物,因此选择细胞色素P450(CYP450)、木质素过氧化物酶(LiP)、漆酶、锰过氧化物酶(MnP)和双加氧酶作为候选酶,并使用硫酸铜、叠氮化钠、硝酸银、乙二胺四乙酸或胡椒基丁醚(PBO)作为酶抑制剂和诱导剂。安排在存在酶抑制剂和诱导剂的情况下对3-PBA及其中间代谢产物的降解以及真菌生物量进行分析,以推断可能的降解酶、共代谢酶和途径。本研究为研究真菌降解3-PBA的共代谢酶提供了一种有前景的方法。•进行了关于3-PBA降解的共代谢酶和途径的MethodsX研究。•研究了呈现酶特性的培养条件。•基于酶的定位、诱导、真菌酶系统以及这些化合物的化学结构对候选酶进行了分析。