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工程化甲酸2-O-硫转移酶的高效表达,具有改善的催化性能

Wuxia Chen^1,2,3,4, Daoan Wang^1,2,3,4, Xintong Xi^1,2,3,4

¹The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China.

Journal of agricultural and food chemistry

|May 14, 2025

概括

研究人员设计了Gallus gallus heparan sulfate 2-O-sulfotransferase (Ga2OST) 酶,显著提高了其表达和催化活性. 这一突破提升了有效的肝素生物制造的潜力.

科学领域:

生物化学生物化学
酵素工程是什么? 酶工程是什么
葡萄糖科学 (Glycoscience) 是一种科学.

背景情况:

肝素的关键生理作用需要高效的生产方法.
肝素硫酸二氧化硫转移酶 (2OST) 是肝素生物合成的关键,但面临表达和活性限制.

研究的目的:

为了提高Gallus gallus衍生的2OST (Ga2OST) 的表达和催化性能,以改善肝素的产生.
为了克服低酶表达和低活性在2OST应用中的挑战.

主要方法:

使用SUMO聚变标签用于活性Ga2OST表达.
利用N-终端同义码子优化来增加表达水平.
构建了一个组合突变物 (SUMO-Ga2OST A98K/Y145F),平衡了催化活性和稳定性.

主要成果:

使用SUMO聚变标签实现了Ga2OST的活性表达.
组合突变SUMO-Ga2OST A98K/Y145F的催化活性增加了2.32倍,半衰期延长了7.80倍.
在5L发酵器中最大化酶活性为5720个单位/毫升,增加了14.79倍,是迄今为止报告的最高活性.

结论:

设计的Ga2OST显示了显著增强的活性表达和催化性能.
开发的突变为可扩展的肝素生物制造提供了坚实的基础.
这项工作为更高效的氨酸相关化合物的工业生产铺平了道路.

关键词:

催化性能催化性能催化性能酶工程是指酶工程的工程.表达水平表达水平的表达水平肝二硫酸二-O-硫酸转移酶的肝二硫酸肝素是一种肝素.硫化硫化剂的使用方法

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