Characterization of the Phosphotransferase from <i>Bacillus subtilis</i> 1101 That Is Responsible for the Biotransformation of Zearalenone
<i>Bacillus</i> microorganisms play an important role in the zearalenone (ZEA) biotransformation process in natural environments. The phosphotransferase pathway in <i>Bacillus</i> is both widespread and relatively well conserved. However, the reaction kinetics of these phosph...
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2025-01-01
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| author | Yuzhuo Wu Qiuyu Zhou Junqiang Hu Yunfan Shan Jinyue Liu Gang Wang Yin-Won Lee Jianrong Shi Jianhong Xu |
| author_facet | Yuzhuo Wu Qiuyu Zhou Junqiang Hu Yunfan Shan Jinyue Liu Gang Wang Yin-Won Lee Jianrong Shi Jianhong Xu |
| author_sort | Yuzhuo Wu |
| collection | DOAJ |
| description | <i>Bacillus</i> microorganisms play an important role in the zearalenone (ZEA) biotransformation process in natural environments. The phosphotransferase pathway in <i>Bacillus</i> 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 <i>Bacillus subtilis</i> 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 (<i>V</i><sub>max</sub>), Michaelis constant (<i>K</i><sub>m</sub>), and catalytic constant (<i>k</i><sub>cat</sub>) were calculated through fitting to be 16.40 μM·s<sup>−1</sup>·mg<sup>−1</sup>, 18.18 μM, and 54.69 s<sup>−1</sup>, respectively. Furthermore, adding 1 mmol/L Fe<sup>2+</sup> or Fe<sup>3+</sup> 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<sup>2+</sup>, suggesting that low concentrations of Fe<sup>2+</sup> or Fe<sup>3+</sup> 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. |
| format | Article |
| id | doaj-art-9a91a4ea25bb4d10ac06685084a2c755 |
| institution | Kabale University |
| issn | 2072-6651 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj-art-9a91a4ea25bb4d10ac06685084a2c7552025-01-24T13:51:13ZengMDPI AGToxins2072-66512025-01-011712110.3390/toxins17010021Characterization of the Phosphotransferase from <i>Bacillus subtilis</i> 1101 That Is Responsible for the Biotransformation of ZearalenoneYuzhuo Wu0Qiuyu Zhou1Junqiang Hu2Yunfan Shan3Jinyue Liu4Gang Wang5Yin-Won Lee6Jianrong Shi7Jianhong Xu8School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaJiangsu 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, ChinaJiangsu 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, ChinaJiangsu 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, ChinaJiangsu 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, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaJiangsu 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, ChinaJiangsu 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, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China<i>Bacillus</i> microorganisms play an important role in the zearalenone (ZEA) biotransformation process in natural environments. The phosphotransferase pathway in <i>Bacillus</i> 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 <i>Bacillus subtilis</i> 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 (<i>V</i><sub>max</sub>), Michaelis constant (<i>K</i><sub>m</sub>), and catalytic constant (<i>k</i><sub>cat</sub>) were calculated through fitting to be 16.40 μM·s<sup>−1</sup>·mg<sup>−1</sup>, 18.18 μM, and 54.69 s<sup>−1</sup>, respectively. Furthermore, adding 1 mmol/L Fe<sup>2+</sup> or Fe<sup>3+</sup> 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<sup>2+</sup>, suggesting that low concentrations of Fe<sup>2+</sup> or Fe<sup>3+</sup> 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.https://www.mdpi.com/2072-6651/17/1/21zearalenonebiotransformation<i>Bacillus subtilis</i>phosphotransferase |
| spellingShingle | Yuzhuo Wu Qiuyu Zhou Junqiang Hu Yunfan Shan Jinyue Liu Gang Wang Yin-Won Lee Jianrong Shi Jianhong Xu Characterization of the Phosphotransferase from <i>Bacillus subtilis</i> 1101 That Is Responsible for the Biotransformation of Zearalenone Toxins zearalenone biotransformation <i>Bacillus subtilis</i> phosphotransferase |
| title | Characterization of the Phosphotransferase from <i>Bacillus subtilis</i> 1101 That Is Responsible for the Biotransformation of Zearalenone |
| title_full | Characterization of the Phosphotransferase from <i>Bacillus subtilis</i> 1101 That Is Responsible for the Biotransformation of Zearalenone |
| title_fullStr | Characterization of the Phosphotransferase from <i>Bacillus subtilis</i> 1101 That Is Responsible for the Biotransformation of Zearalenone |
| title_full_unstemmed | Characterization of the Phosphotransferase from <i>Bacillus subtilis</i> 1101 That Is Responsible for the Biotransformation of Zearalenone |
| title_short | Characterization of the Phosphotransferase from <i>Bacillus subtilis</i> 1101 That Is Responsible for the Biotransformation of Zearalenone |
| title_sort | characterization of the phosphotransferase from i bacillus subtilis i 1101 that is responsible for the biotransformation of zearalenone |
| topic | zearalenone biotransformation <i>Bacillus subtilis</i> phosphotransferase |
| url | https://www.mdpi.com/2072-6651/17/1/21 |
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