Mechanistic model for the synthesis of N-acetylneuraminic acid using N-acetylneuraminate lyase from Escherichia coli K12
Mechanistic model for the synthesis of N-acetylneuraminic acid using N-acetylneuraminate lyase from Escherichia coli K12
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DOI：
10.1016/j.molcatb.2012.05.016
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发表时间：
2012-11-01
影响因子：
--
通讯作者：
Hoelsch, Kathrin
中科院分区：
文献类型：
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作者：
Groher, Anna;Hoelsch, Kathrin
大肠杆菌K12的N-乙酰神经氨酸裂解酶（NAL）是生产N-乙酰神经氨酸（Neu 5Ac）的重要酶，催化N-乙酰-D-甘露糖胺（ManNAc）与丙酮酸的可逆羟醛缩合反应。尽管这种酶的工业重要性，其动力学机制从来没有被阐明。初始速率模式与丙酮酸盐首先结合的快速平衡有序双单机制一致。基于过程曲线分析法，建立了Neu 5Ac合成反应过程的机理模型。该模型在较宽的初始条件范围内准确地再现了实验数据。正确分配的动力学机制是一个关键因素，在优化酶的合成，通过数学模型，这已成为不可或缺的工具，成本效益的生物催化过程的设计。(C)出版社：Elsevier B. V.
N-Acetylneuraminate lyase (NAL) from Escherichia coli K12 is an important enzyme for the production of N-acetylneuraminic acid (Neu5Ac), catalyzing the reversible aldol condensation between N-acetyl-D-mannosamine (ManNAc) and pyruvate. Despite the industrial importance of this enzyme, its kinetic mechanism has never been elucidated before. The initial rate patterns were consistent with a rapid-equilibrium ordered bi uni mechanism with pyruvate binding first. Based on progress curve analysis, a mechanistic model was developed to predict the reaction course of Neu5Ac synthesis. The model accurately reproduced the experimental data in a wide range of initial conditions. The correct assignment of the kinetic mechanism is a critical element in optimizing enzymatic syntheses by means of mathematical models, which have become indispensable tools for the design of cost-effective biocatalytic processes. (C) 2012 Published by Elsevier B.V.