通过平衡子单元的表达方式,显著增强酸酸酶的催化活性
Yi Guo1,2, Jiaxin Li1,2, Song Zhang3
1School of Chemical Engineering, Laboratory of Advanced Materials and Catalytic Engineering, Dalian University of Technology, Dalian, 116024, China.
Chembiochem : a European journal of chemical biology
|December 1, 2024
概括
在大肠杆菌 (E. coli) 中优化mRNA结构改善了ReNHase表达和活性. 融合标签增强了可溶性表达,但影响了亚单元平衡和酶功能.
科学领域:
- 生物技术是生物技术.
- 蛋白质工程是指蛋白质工程.
- 微生物发酵 微生物发酵
背景情况:
- 大肠杆菌 (E. coli) 是重组蛋白质生产的一个关键系统.
- 之前的研究已经确定了具有催化活性的重组Rhodococcus erythropolis CCM2595化酶 (ReNHase).
- 不平衡的子单元表达阻碍了异构的ReNHase的功能组合.
研究的目的:
- 为了提高ReNHase的翻译效率和子单位表达平衡.
- 使用聚变标签来改善ReNHase的可溶性表达.
- 调查聚变标签对ReNHase子单元平衡和活动的影响.
主要方法:
- 优化了ReNHaseβ子单元的核糖体结合部位中的mRNA二次结构.
- 在大肠杆菌中构建并表达了ReNHase与优化的mRNA和各种融合标签.
- 评估了酶活性,可溶性表达水平和子单位表达平衡.
主要成果:
- 优化的mRNA翻译效率导致ReNHase特定活性增加了12倍.
- 融合标签改善了可溶性ReNHase的表达,但破坏了亚单元平衡和减少了酶活性.
- 二级结构优化策略成功地平衡了α和β子单元表达.
结论:
- 优化mRNA二级结构是一种有效的策略,可以改善异质多子单元蛋白质的表达.
- 融合标签可以增强可溶性表达,但需要仔细选择,以避免对酶功能的负面影响.
- 这项研究为优化复杂的多子单元酶的生产提供了有价值的方法.
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