2-(2-Oxopyrrolidin-1-yl)-acetic acid(2-OAA)is an important precursor for the synthesis of piracetam and plays a significant role in the field of pharmaceutical synthesis.However,there is currently a lack of biological methods and reaction mechanism analysis for synthesizing it.To address this research gap,based on the principle of similarity reaction,the β-lactam synthetase(SfAsnA)from Streptomyces fulvorobeus was identified.This enzyme has the ability to synthesize 2-OAA in the aqueous phase,exhibiting an enzyme activity of 109.8 U/g and a total conversion number(TTN)of up to 79.02.Subsequently,the protein model construction of the SfAsnA was analyzed with the substrate to determine the combination of the substrate in the enzyme activity center.Next,the reaction mechanism of the non-enzymatic reaction for synthesizing 2-OAA was analyzed in detail to verify the chemical synthesis mechanism and relative energy profile of the target reaction.Finally,the enzymatic response mechanism is studied,and mutation verification is used to catalyze dual-combined bodies(Y254 and E288)to further determine the SfAsnA catalytic 2-OAA synthesis mechanism and the key role of the dual-connected body in the target response.It provides a solid theoretical foundation for enzyme synthesis of 2-OAA.
关键词
2-(氧代吡咯烷-1基)-乙酸/生物催化/内酰胺化/分子模拟/反应能垒/计算化学
Key words
2-(2-oxopyrrolidin-1-yl)-acetic acid/biocatalysis/lactamization/molecular simulation/reaction energy barrier/computational chemistry
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