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理性设计的l-氨酸转氨酸酶介导系统,以提高佛罗芬尼科尔中间体的生产

Zhiwen Xi¹, Lihong Li¹, Zhiyong Liu¹

¹Lab of Brewing Microbiology and Applied Enzymology, School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, P. R. China.

Journal of agricultural and food chemistry

|December 28, 2023

概括

工程酶有效合成l-threo-p-methylsulfonylphenylserine (化合物1b),这是一个关键的花尼醇中间体. 这项研究提供了一种可持续的方法来生产具有高产量和选择性的1b.

科学领域:

生物催化和酶工程生物催化和酶工程
有机合成有机合成
制药化学制药化学制药化学

背景情况:

佛罗芬尼科尔是一种重要的抗生素,其合成依赖于l-threo-p-methylsulfonylphenylserine (化合物1b) 作为一个关键的中间体.
化合物1b的高效和可持续合成对于制药生产至关重要.

研究的目的:

理性地设计和工程Burkholderia扩散l-threonine转化酶 (BuLTTA) 来增强化合物1b的合成.
开发一种合酶系统,以提高化合物1b生产的产量和选择性.

主要方法:

基于序列结构功能关系的BuLTTA的合理酶设计.
局部导向的突变生成产生M4变体 (Asn35Ser/Thr352Asn).
分子动力学模拟以阐明催化机制.
通过与K. kurtzmanii的酒精脱酶 (KkADH) 和辅因子再生系统的合工程BuLTTA.

主要成果:

M4 BuLTTA突变物获得了35.5毫米的化合物1b,转化率为88.8%,立体选择性为93.8%.
分子动力学揭示了M4活性部位的增强相互作用,改善了催化作用.
结合的酶系统产生了115.2mM的1b,其转换率为96%和95.5%的二选择性.

结论:

设计的BuLTTA (M4变种) 显著增强了化合物1b的合成.
与辅助因子再生相结合的生物催化系统为化合物1b的生产提供了高效和可持续的途径.
这一战略为工业合成花尼科尔中间体提供了一个有前途的方法.

关键词:

副产品清除系统副产品清除系统辅助因子回收系统的系统.l-threo-p-methylsulfonylphenylserine的使用情况一个L-threonine转氨酸酶.分子动力学模拟模拟理性的设计理性的设计.

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