Dramatically Improved Performance of an Esterase for Cilastatin Synthesis by Cap Domain Engineering
| العنوان: | Dramatically Improved Performance of an Esterase for Cilastatin Synthesis by Cap Domain Engineering |
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| المؤلفون: | Hui-Lei Yu, Zheng-Jiao Luan, Qi Chen, Jian-He Xu, Bao-Di Ma, Yi-Ke Qi |
| المصدر: | Industrial & Engineering Chemistry Research. 55:12167-12172 |
| بيانات النشر: | American Chemical Society (ACS), 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, chemistry.chemical_classification, Cilastatin, 010405 organic chemistry, General Chemical Engineering, Mutant, Wild type, Substrate (chemistry), General Chemistry, 01 natural sciences, Esterase, Industrial and Manufacturing Engineering, 0104 chemical sciences, 03 medical and health sciences, 030104 developmental biology, Enzyme, chemistry, Biochemistry, Hydrolase, medicine, Specific activity, medicine.drug |
| الوصف: | Whole-protein random mutation and substrate tunnel evolution have recently been applied to the pharmaceutically relevant esterase RhEst1 for the synthesis of a cilastatin precursor. The mutant RhEst1M1 (=RhEst1A147I/V148F/G254A) was identified from a large library consisting of 1.5 × 104 variants. Though the activity of this mutant was improved 5-fold, the enantioselectivity for biohydrolysis decreased at the same time. Herein a smart library (3.0 × 103) focused on the cap domain of RhEst1 was constructed to improve its catalytic performance comprehensively. As a result, a variant designated as RhEst1M2 (=RhEst1M1-A143T), showed a 6-fold increase in specific activity compared with the wild type. Meanwhile, the decreased enantioselectivity for enzymatic resolution was recovered to the native enzyme level. The melting temperature of RhEst1M2 was nearly 11 °C higher than that of the wild type. This work provides detailed insight into the vital role of α/β hydrolase cap domains in influencing all aspects of e... |
| تدمد: | 1520-5045 0888-5885 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::9a7a0d38703e00eae646c71b8bba17fe https://doi.org/10.1021/acs.iecr.6b02440 |
| رقم الأكسشن: | edsair.doi...........9a7a0d38703e00eae646c71b8bba17fe |
| قاعدة البيانات: | OpenAIRE |
Find this issue from the American Chemical Society | تدمد: | 15205045 08885885 |
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