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通过定向进化增强两个微生物尿酶的催化效率

Xiaoyuan Tang¹, Liling Qin¹, Yuze Xia¹

¹Department of Biological Medicines & Shanghai Engineering Research Center of Immunotherapeutics, Fudan University School of Pharmacy, 826 Zhangheng Road, Pudong, Shanghai 201203, PR China.

International journal of biological macromolecules

|February 1, 2025

概括

研究人员从Aspergillus flavus和Candida utilis中设计了新型尿酶突变体,实现了潜在的高尿血和痛风治疗的创纪录的高酶活性.

科学领域:

生物化学生物化学
酶学是一种酶学.
代谢障碍代谢障碍代谢障碍

背景情况:

尿酶在氨酸代谢中至关重要,它将尿酸转化为阿兰托因.
它用于治疗高尿血和痛风治疗.
增强尿酶活性对于改善治疗疗效至关重要.

研究的目的:

为了产生和描述具有显著增强酶活性的尿酶突变体.
为了确定负责增加尿酶功能的关键突变.
为了评估工程尿的体内治疗潜力.

主要方法:

利用易出错的PCR和高通量查用于突变基因的产生.
采用位点定向突变发生法来确认关键的氨基酸替代.
进行了体内实验以评估临床适用性.

主要成果:

开发了Aspergillus flavus尿酶突变 (af-UA^M) 的46.21U/mg活动.
开发出Candida utilis uricase突变 (cu-UA^M) 具有31.43U/mg的活性.
确定了Thr231Ala和Val234Met的替代物,这些替代物对于增强活性至关重要.

结论:

通过蛋白质工程实现了迄今为止报告的最高尿酶活性.
建立了结构-功能关系,以改进尿病例设计.
在治疗高尿血症方面显示出显著的治疗潜力.

关键词:

定向进化是指导进化的.尿酸是什么尿酸是什么乌里卡赛斯 (Uricase) 是一种

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