通过高通量计算工程增强双域氨基酶的冷适应
Ning Ding1, Yaoyukun Jiang1, Robbie Ge1
1Department of Chemistry, Vanderbilt University, Nashville, TN, 37235, USA.
Angewandte Chemie (International ed. in English)
|April 24, 2025
概括
适应寒冷的酶可以提高工业的可持续性. 研究人员确定了域分离是粉酶适应寒冷的关键,使其在低温下增强活性.
科学领域:
- 生物化学 生物化学
- 酶工程是什么? 酶工程是什么?
- 结构生物学 结构生物学
背景情况:
- 适应寒冷的酶通过减少能源使用和排放,为工业可持续性带来了好处.
- 酶冷适应的基础分子机制尚不清楚,这限制了合理的设计.
- 双域酶,像粉酶一样,为设计冷活性变体提供了机会.
研究的目的:
- 阐明双域酶冷适应的分子基础.
- 开发一种工程冷适应性粉酶的策略.
- 为了研究在低温下酶功能中域间分离的作用.
主要方法:
- 使用分子动力学 (MD) 模拟,对适应寒冷的Saccharophagus降解氨基酶 (sdA) 和 mesophilic Pseudomonas saccharophila氨基酶 (psA) 的比较分析.
- 生物化学测试以验证低温下酶活性.
- 使用新型域分离指数的新型酶变体的in silico选.
主要成果:
- 与psA相比,冷适应的sdA在低温下显示其催化域 (CD) 和碳水化合物结合模块 (CBM) 之间的域间分离更大.
- 开发了一种新的指标,即域分离指数,以指导酶变异查.
- 改造后的psA变种psA121在0°C时相对活性增加了3倍.
- MD模拟表明,psA121中的螺旋连接器促进域间分离和动态化,以增强低温催化.
结论:
- 域间分离是一个关键因素,有助于双域氨基酶的寒冷适应.
- 域分离指数为冷适应酶的in silico选和工程提供了有价值的工具.
- 这项研究为引入其他酶系统的冷适应提供了一个框架,推进可持续的工业过程.
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