Wu Yuzhuo, Zhou Qiuyu, Hu Junqiang, Shan Yunfan, Liu Jinyue, Wang Gang, Lee Yin-Won, Shi Jianrong, Xu Jianhong
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China.
Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Key Laboratory for Agro-Product Safety Risk Evaluation (Nanjing), Ministry of Agriculture and Rural Affairs, Key Laboratory for Control Technology and Standard for Agro-Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Collaborative Innovation Center for Modern Grain Circulation and Safety, Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
Toxins (Basel). 2025 Jan 3;17(1):21. doi: 10.3390/toxins17010021.
microorganisms play an important role in the zearalenone (ZEA) biotransformation process in natural environments. The phosphotransferase pathway in is both widespread and relatively well conserved. However, the reaction kinetics of these phosphotransferases remain poorly understood, and their catalytic activities are suboptimal. In this study, a ZEA phosphotransferase, ZPH1101, was identified from 1101 using genome sequencing. The product transformed by ZPH1101 was identified as phosphorylated ZEA (ZEA-P) through LC-TOF-MS/MS analysis. The experiments conducted on MCF-7 cells demonstrated that ZEA-P exhibited a lower level of estrogenic toxicity than ZEA. The optimal reaction conditions for ZPH1101 were determined to be 45 °C and pH 8.0. The maximum velocity (), Michaelis constant (), and catalytic constant () were calculated through fitting to be 16.40 μM·s·mg, 18.18 μM, and 54.69 s, respectively. Furthermore, adding 1 mmol/L Fe or Fe to the reaction system increased the efficiency of ZPH1101 in converting ZEA by 100% relative to the system containing solely 1 mmol/L ATP and 1 mmol/L Mg, suggesting that low concentrations of Fe or Fe can improve the ZPH1101-mediated transformation of ZEA. This study contributes to the enzymatic removal of ZEA and broadens the spectrum of strain and enzyme options available to researchers for ZEA detoxification efforts.
微生物在自然环境中的玉米赤霉烯酮（ZEA）生物转化过程中发挥着重要作用。 中的磷酸转移酶途径广泛存在且相对保守。然而，这些磷酸转移酶的反应动力学仍知之甚少，其催化活性也不理想。在本研究中，通过基因组测序从1101中鉴定出一种ZEA磷酸转移酶ZPH1101。通过LC-TOF-MS/MS分析确定ZPH1101转化的产物为磷酸化ZEA（ZEA-P）。在MCF-7细胞上进行的实验表明，ZEA-P的雌激素毒性水平低于ZEA。确定ZPH1101的最佳反应条件为45℃和pH 8.0。通过拟合计算出最大速度（）、米氏常数（）和催化常数（）分别为16.40μM·s·mg、18.18μM和54.69 s。此外，相对于仅含有1 mmol/L ATP和1 mmol/L Mg的体系，向反应体系中添加1 mmol/L Fe或Fe可使ZPH1101将ZEA转化的效率提高100%，这表明低浓度的Fe或Fe可改善ZPH1101介导的ZEA转化。本研究有助于ZEA的酶促去除，并拓宽了研究人员在ZEA解毒工作中可用的菌株和酶的选择范围。
