| Autor: |
Xi, Zhiwen, Li, Lihong, Liu, Zhiyong, Wu, Xiaolong, Xu, Yan, Zhang, Rongzhen |
| Zdroj: |
Journal of Agricultural and Food Chemistry; January 2024, Vol. 72 Issue: 1 p461-474, 14p |
| Abstrakt: |
l-threo-p-methylsulfonylphenylserine (compound 1b) is the main intermediate of florfenicol, and its efficient synthesis has been the subject of current research. Herein, Burkholderia diffusal-threonine transaldolase (BuLTTA) was rationally designed based on the sequence–structure–function relationship. A mutant M4 (Asn35Ser/Thr352Asn) could produce 35.5 mM 1bwith 88.8% conversion and 93.8% diastereoselectivity, 314 and 129% of the values observed for wild-type BuLTTA. Molecular dynamics simulations indicated that the shortened distance between key active site residues and the transition state (PLP-1b) and the improved hydrogen bond force enhanced the catalytic performance of the M4 variant. Then, the mutant M4 was combined with K. kurtzmaniialcohol dehydrogenase (KkADH) to eliminate the BuLTTA-inhibiting byproduct acetaldehyde, and a cosubstrate was added to regenerate the ADH cofactor NADH. Under optimized conditions, the yield of 1breached 115.2 mM with a conversion of 96% and a diastereoselectivity of 95.5%. This work provides a new strategy for the efficient and sustainable production of 1b. |
| Databáze: |
Supplemental Index |
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