溶解剂诱导的脂酶自发重新折叠
Cheng Cheng1, Yongqin Su2, Lupeng Cui3
1School of Pharmaceutical Sciences, Nanjing Tech University, No. 30 Puzhu South Road, Nanjing, 211816, China. Cheng2@njtech.edu.cn.
Applied microbiology and biotechnology
|January 6, 2026
概括
这项研究引入了一种用于溶性蛋白质生产和重新折叠的新策略,使用糖醇,DMSO和TMAO等辅溶剂. 甘油显著增强了脂酶活性,为蛋白质重新折叠提供了更有效的方法.
科学领域:
- 生物化学 生物化学
- 蛋白质工程是指蛋白质工程.
- 计算生物学 计算生物学
背景情况:
- 大肠杆菌中的蛋白质过度表达往往导致不活跃的包容体 (IBs),需要劳动密集的溶解和重新折叠.
- 从IB中获得生物活性蛋白质对于各种应用至关重要.
- 开发有效的可溶性蛋白质生产和重新折叠的战略是一个重大挑战.
研究的目的:
- 开发一种用于Burkholderia ambifaria lipase YCJ01.01的可溶性生产和重新折叠的战略.
- 研究辅溶剂对可溶性脂酶YCJ01.01的活性的影响.
- 用分子动力学模拟来阐明辅溶剂诱导的蛋白质重新折叠的机制.
主要方法:
- 融合标签应用在E. coli中丰富的可溶性表达脂酶YCJ01的应用.
- 用各种辅溶剂 (糖醇,DMSO,TMAO) 化可溶性脂酶以评估活性.
- 分子动力学 (MD) 模拟以了解辅溶剂-蛋白质相互作用和重新折叠机制.
主要成果:
- 可溶性脂酶YCJ01表现出部分活性,该活性被辅溶剂显著增强.
- 脂酶活性达到1003U/mL与25%的糖醇,754U/mL与15%的DMSO,和501U/mL与4M的TMAO.
- MD模拟显示,糖醇和DMSO通过与疏水区域相互作用直接稳定脂酶,而TMAO通过加强水与水的相互作用来间接稳定.
结论:
- 辅溶剂,特别是甘油,有效增强可溶性脂酶YCJ01.01的活性.
- 该研究提供了对辅助溶剂诱导的蛋白质稳定直接和间接机制的见解.
- 这种方法为高效的可溶性蛋白质生产和重新折叠提供了一个有希望的策略.
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