Enhanced α-Zearalenol Hydrolyzing Activity of a Mycoestrogen-Detoxifying Lactonase by Structure-Based Engineering
| Autor: | Huazhong Li, Weidong Liu, Jian Jin, Chen Yun, Tzu-Ping Ko, Guizhi Liu, Wei Peng, Rey-Ting Guo, Ya-Shan Cheng, Qian Li, Yingying Zheng, Wenting Liu, Zhongxia Xu, Jian-Wen Huang, Chun-Chi Chen |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification 030102 biochemistry & molecular biology biology Chemistry Stereochemistry Rational design Substrate (chemistry) Mycoestrogen General Chemistry Catalysis 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology Enzyme Biochemistry Catalytic triad Hydrolase Lactonase biology.protein Specific activity |
| Zdroj: | ACS Catalysis. 6:7657-7663 |
| ISSN: | 2155-5435 |
| DOI: | 10.1021/acscatal.6b01826 |
| Popis: | The enzyme ZHD101 from Clonostachys rosea hydrolyzes and deactivates the mycotoxin zearalenone (ZEN) and its zearalenol (ZOL) derivatives. ZHD101 prefers ZEN to ZOL as its substrate, but ZOL, especially the α-form, shows higher estrogenic toxicity than ZEN. To enhance α-ZOL selectivity, we solved the complex structures of ZHD101 with both ZOLs and modified several lactone-surrounding residues. Among the mutants, V153H maintained activity for ZEN but showed a 3.7-fold increase in specific activity against α-ZOL, with an ∼2.7-fold reduction in substrate affinity but a 5.2-fold higher turnover rate. We then determined two V153H/ZOL complex structures. Here, the α-ZOL lactone ring is hydrogen-bonded to the H153 side chain, yielding a larger space for H242 to reconstitute the catalytic triad. In conclusion, structure-based engineering was successfully employed to improve the ZHD101 activity toward the more toxic α-ZOL, with great potential in further industrial applications. |
| Databáze: | OpenAIRE |
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