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在产品释放裂中工程聚糖化酸,以改变其产品配置

Meixing Wang¹, Yufan Xian¹, Zhenghui Lu²

¹College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.

International journal of biological macromolecules

|November 29, 2023

概括

在neopullulase Amy117中改变单个氨基酸残留物改变了其酶活性,使得pullulan的特定水解成为panose. 这一发现为处理多糖的酶工程提供了洞察力.

科学领域:

酶学是一种酶学.
生物化学生物化学
结构生物学结构生物学

背景情况:

多糖化酶是生产工业应用的寡糖的关键酶.
控制酶化物组成的精确机制尚未完全理解.
Neopullulase Amy117独特地将pullulan水解成panose,与产生混合产品的同源酶不同.

研究的目的:

通过新酶Amy117.7研究pullulan的特异性水解的结构基础.
了解为什么高度同源的酶会产生不同的水解化合物组成.
通过基于结构的修改来设计酶活性.

主要方法:

艾米117和马尔托氨酶ThMA的比较序列和结构分析.
通过改变一个关键残留物 (Amy117中的Thr299) 来进行基于结构的合理设计.
分子对接模拟以分析基质-酶相互作用和产品释放.
长链普卢兰 (G8) 形状的计算建模.

主要成果:

一个单一的残留物,Thr299在Amy117中 (His294在ThMA中),被确定为对产品特异性的关键.
突变分析成功切换了Amy117和ThMA之间的产品配置文件.
分子对接显示,关键的残留物影响了板释放率.
建模表明,pullulan G8在酶的裂中经历了构造变化,促进了特定的α-1,6-糖化合物键的识别.

结论:

特定的残留物Thr299决定了神经 Amy117.7 的独特的水解特异性.
通过改变关键残留物,酶工程可以精确地控制寡糖类产品概况.
了解酶活性部位内的基质构造变化对于酶设计至关重要.

关键词:

聚糖化酸盐的多糖化酸盐.产品简介产品简介一个单点突变的突变.

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