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从适应寒冷的Variovorax sp.中探索新型特雷哈拉酶. PAMC28711:功能性特征的描述

Prasansah Shrestha¹, Jayram Karmacharya², Ki-Hwa Kim¹

¹Department of Life Science and Biochemical Engineering, Graduate School, SunMoon University, Asan 31460, South Korea; Genome-based Bio-IT Convergence Institute, Asan 31460, South Korea.

International journal of biological macromolecules

|May 20, 2024

概括

这项研究确定了Variovorax sp.中的两个新型三酶 (GH15和GH37). PAMC28711,详细介绍了它们的不同活动,催化位点和在乙醇增强和抑制剂查中的潜在应用.

科学领域:

微生物学微生物学
酶学是一种酶学.
生物化学生物化学

背景情况:

三酶酶对于细菌中的三糖代谢至关重要.
在单个细菌菌株内存在多个三酶基因是罕见的,以前没有报告过.
这种植物名为Variovorax sp. PAMC28711是一种适应寒冷的细菌,它包含两个不同的特雷酶家族:GH15和GH37.

研究的目的:

描述来自Variovorax sp.的两个新型三酶的特征. 这就是PAMC28711.11.
阐明它们的基质特异性,催化机制和生物化学性质.
探索这些特雷哈拉酶的潜在应用.

主要方法:

GH15和GH37三酶的基因鉴定和特征.
使用α,α-三糖进行基质特异性测定.
位点定向的突变发生,以确定关键的催化残留物.
在各种条件下 (pH,乙醇) 进行酶活性测定和稳定性测试.
用validoxylamine A.进行抑制活性测定.

主要成果:

两种GH37 (Vtre37) 和GH15 (Vtre15) 三酶都对α,α-三糖表现出水解活性.
确定了关键的催化部位:Vtre37中的D280和E469;Vtre15中的E389和E554.
与Vtre15 (408.39 μmolmg-1) 相比,Vtre37表现出明显更高的酶活性 (1306.33 μmolmg-1).
Vtre37在酸性条件下表现出显著的稳定性和对10%乙醇的耐药性.
瓦利多克西胺A强烈抑制了Vtre37三酶,其Ki值为16.85nM.

结论:

对Vtre37和Vtre15的表征增强了对细菌三糖代谢的理解,以及多个三糖基因的生物学意义.
Vtre37显示了作为乙醇增强剂的使用潜力.
在三醇降解途径内,Vtre37可以成为设计抑制剂的目标.

关键词:

催化机制是一种催化机制.适应寒冷的适应者树木气息 (Trehalase) 是一种树木气息.

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