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通过转录组分析鉴定参与玉米赤霉烯酮解毒的 H6 硫酯酶。

Identification of a H6 Thioesterase Involved in Zearalenone Detoxification by Transcriptomic Analysis.

机构信息

College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450000, China.

College of Environmental and Resource Sciences, Henan Agricultural University, Zhengzhou 450000, China.

出版信息

J Agric Food Chem. 2020 Sep 16;68(37):10071-10080. doi: 10.1021/acs.jafc.0c03954. Epub 2020 Sep 1.

Abstract

Zearalenone (ZEA), a nonsteroidal estrogenic mycotoxin produced by , induces hyperestrogenic responses in mammals and can cause reproductive disorders in farm animals. In this study, a transcriptome analysis of H6, which was previously identified as a ZEA-degrading bacterium, was conducted with high-throughput sequencing technology, and the differentially expressed genes were subjected to gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses. Among the 16 upregulated genes, BAMF_RS30125 was predicted to be the key gene responsible for ZEA degradation. The protein encoded by BAMF_RS30125 was then expressed in , and this recombinant protein (named ZTE138) significantly reduced the ZEA content, as determined by the enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC), and decreased the proliferating activity of ZEA in MCF-7 cells. What is more, the liquid chromatography-tandem mass spectrometry (LC-MS/MS) results showed that the relative molecular mass and the structure of ZEA also changed. Sequence alignment of the ZTE138 protein showed that it is a protease that belongs to the YBGC/FADM family of coenzyme A thioesterases, and thus, the protein can presumably cleave the ZEA lactone bond and break down its macrolide ring.

摘要

玉米赤霉烯酮(ZEA)是一种由 产生的非甾体类雌激素真菌毒素,它会引起哺乳动物的雌激素过度反应,并可能导致农场动物的生殖障碍。在这项研究中,我们利用高通量测序技术对先前被鉴定为 ZEA 降解菌的 H6 进行了转录组分析,对差异表达基因进行了基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析。在 16 个上调基因中,预测 BAMF_RS30125 是负责 ZEA 降解的关键基因。然后,在 中表达了 BAMF_RS30125 编码的蛋白,该重组蛋白(命名为 ZTE138)通过酶联免疫吸附测定(ELISA)和高效液相色谱(HPLC)显著降低了 ZEA 的含量,并降低了 ZEA 在 MCF-7 细胞中的增殖活性。更重要的是,液相色谱-串联质谱(LC-MS/MS)结果表明,ZEA 的相对分子质量和结构也发生了变化。ZTE138 蛋白的序列比对表明,它是一种属于辅酶 A 硫酯酶的 YBGC/FADM 家族的蛋白酶,因此,该蛋白可能会切断 ZEA 的内酯键并破坏其大环。

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