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通过以结构为基础的烯合并来提高脂酶C的热稳定性,以提高其脱性能

Yiling Tang¹, Zhiwei Lin¹, Lei Ren¹

¹School of Food Science and Engineering, South China University of Technology, Guangzhou, 510640, People's Republic of China.

Applied biochemistry and biotechnology

|May 15, 2025

概括

将普罗林引入真菌脂酶C (TiPLC) 显著提高了其热稳定性和活性. 这种蛋白质工程方法提高了工业应用的酶性能,例如油脱皮.

科学领域:

酶学是一种酶学.
蛋白质工程是指蛋白质工程.
生物技术是生物技术.

背景情况:

来自Talaromyces islandicus的真菌脂酶C (TiPLC) 具有较差的热稳定性,限制了其工业应用.
蛋白质的热稳定性对于在各种反应条件下酶的性能至关重要.

研究的目的:

为了提高来自Talaromyces islandicus的真菌脂酶C (TiPLC) 的热稳定性.
为了研究proline合并对TiPLC在油脱过程中的稳定性,活性和性能的影响.

主要方法:

在Pichia pastoris中TiPLC的异质表达和生物化学表征.
基于结构的普罗林纳入策略,产生单位 (E92P,A375P) 和双位 (E92P-A375P) 突变物.
酶活性测定,热稳定性测量 (t1/2) 和油脱实验.
分子动力学模拟以了解增强热稳定性的结构基础.

主要成果:

单位突变物E92P和A375P表现出更好的热稳定性 (分别在40°C时长达1.62倍和1.27倍) 和20%的活性增加.
与野生类型相比,双重突变E92P-A375P的热稳定性增加了2.43倍.
双重突变显著提高了油脂脱效率,将残留减少到78ppm (与野生类型的131ppm相比).
分子模拟证实,proline的加入增加了柔性区域的刚性,有助于增强热稳定性.

结论:

结构导向的普罗林结合是一种有效的策略,可以提高真菌脂酶C的热稳定性和性能.
增强的TiPLC突变体显示出需要高温酶活性的工业应用的潜力,例如油脱.
这项研究提供了通过氨酸替代蛋白质热稳定背后的分子机制的见解.

关键词:

塔拉罗米切斯 (Talaromyces) 是一个岛屿动物.酵素脱皮是一种酶性脱皮.脂酶C是一种脂酶.素的结合物素的结合热稳定性热稳定性

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