通过使用循环替换,改善来自Bacillus RN.1的酸酶的最佳pH值和特异活性
Piwu Li1,2, Xiaofeng Wei2, Yun Wang2
1State Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology, Jinan, Shandong, China.
Frontiers in bioengineering and biotechnology
|July 20, 2023
概括
工程性乳酸酶 (BspPel-th) 显示了增强的活性和抗性,使其适合工业应用. 这种改进的酶在广泛的pH范围内表现出显著的稳定性.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 蛋白质工程是指蛋白质的工程.
背景情况:
- 性乳酸酶对于造纸和废水处理等行业至关重要.
- 目前的局限性包括低活性和低抗性,阻碍工业用途.
- 研究了Bacillus RN.1的乳酸酶 (BspPel),以寻找潜在的改善.
研究的目的:
- 为了增强性乳酸酶 (BspPel) 的活性和抗性.
- 探索特定突变,如R260在酶特性中的作用.
- 评估修改酶的工业适用性.
主要方法:
- 在大肠杆菌BL21 (DE3) 中BspPel的异质表达.
- 通过循环替换进行酶工程,以提高活性和抗性.
- 生物化学测试和分子动力学模拟以表征酶.
主要成果:
- 工程BspPel-th在60°C和pH值11.0时表现出最佳的活性.
- 与野生类型相比,特定酶活性增加了4.4倍,达到139.4U/mg.
- BspPel-th在pH值3.0-11.0之间表现出显著的稳定性,并改善了循环的灵活性.
结论:
- 修改后的BspPel-th显示了显著增强的性能特征.
- 它的高活性和性pH的稳定性表明它具有强大的工业潜力.
- 这种工程酶是高pH工业过程的有希望的候选者.
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