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从Enterobacter faecalis SK32.001通过组合性突变发生的过程中增强了 arginine deiminase的热稳定性和催化活性

Mengli Li¹, Yijing Zhang¹, Tao Zhang²

¹State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi, Jiangsu 214122, China.

International journal of biological macromolecules

|November 25, 2024

概括

理性设计增强了Enterococcus faecalis arginine deiminase (ADI),以提高稳定性和活性. 一种5分突变体的特异性活性增加了两倍,点上升了11.62°C,显示出工业应用的潜力.

科学领域:

生物化学生化学
酶工程是什么? 酶工程是什么?
蛋白质工程是指蛋白质工程.

背景情况:

氨酸减轻酶 (ADI) 具有潜在的临床和工业用途,但其热稳定性和催化效率较低.
优化ADI的特性对于扩展其应用至关重要.

研究的目的:

为了合理设计Enterococcus faecalis arginine deiminase (ADI),以提高热稳定性和催化活性.
为了确定在生理条件下改善ADI性能的特定突变.

主要方法:

通过堆叠单点突变来构建ADI的120个组合突变.
评估突变体的热稳定性 (Tm) 和酶活性.
使用AlphaFold 2进行结构预测,以了解性能改进的基础.

主要成果:

突变S604,S700,S601和S606显示出更高的化温度 (Tm).
突变S605,S547,S602,S607,S517和S557表现出增强的酶活性.
五点突变S547表现出显著的pH耐受性 (pH4.5-9.5),两倍高的特异活性 (131.60U/mg),以及Tm (64.04°C) 的11.62°C增加.

结论:

理性设计策略可以有效地优化微生物酶特性.
增强的ADI突变体显示出工业应用的巨大潜力.
结构洞察力表明,增加的刚性,新的键和更多的疏水性残留物有助于提高酶的性能.

关键词:

氨酸减小酶的作用催化活动是催化活动.菌 (Enterococcus faecalis) 是一种菌.酶工程是什么?酶工程是什么?热稳定性热稳定性

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