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金属β-乳酸酶SMB-1在美罗烯的选择性压力下演变为更高效的酶

Xiaoting Dong¹, Wenli Liu¹, Yuxuan Dong¹

¹College of Life Science, Northwest University, Xi'an 710069, China.

Journal of inorganic biochemistry

|July 21, 2023

概括

金属β-乳酸酶 (MβLs) 驱动卡巴胺抗生素耐药性. 研究SMB-1进化发现了增强酶灵活性,稳定性和活性的突变,导致药物耐药性增加.

科学领域:

微生物学微生物学
分子生物学分子生物学
生物化学生物化学

背景情况:

金属β-乳酸酶 (MβLs) 是一个重大威胁,它对卡巴胺抗生素产生了耐药性.
了解MBL的演变对于对抗抗生素耐药性至关重要.

研究的目的:

为了研究来自Serratia marcescens的SMB-1金属β-乳糖酶的突变和演变.
为了阐明基因和结构变化背后的碳烯耐药性.

主要方法:

使用不连续的美罗胺度梯度的微生物进化实验.
在SMB-1中分析点突变,双位点突变和移突变.
确定最小抑制度 (MIC) 和催化效率 (kcat/Km).
活跃站点变化的结构分析和循环灵活性.

主要成果:

在SMB-1.1的C端发现了关键突变 (A242G,S257R,C240G/E258G,M5,M12,M13).
观察到美罗MIC (8-120倍) 和催化效率 (365-647%) 的显著增加.
检测到的结构变化包括活性部位障碍和增强的循环1灵活性,增加腔体积和Zn2+部位暴露.

结论:

基因突变推动了SMB-1的进化,增强了它的灵活性,稳定性和水解活性.
这些适应性有助于持续的耐药性和对卡巴胺抗生素的耐药性.
这项研究提供了对抗生素耐药性进化的分子机制的见解.

关键词:

抗生素耐药性抗生素耐药性规范性变化规范性变化距离突变距离突变进化进化进化进化进化进化进化金属β-乳酸酶SMB-1的使用突变效应的影响.

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