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