宁夏地区牛源金黄色葡萄球菌β-内酰胺酶的分析及作用方式的研究

doi:10.11843/j.issn.0366-6964.2020.05.025

摘要/Abstract

摘要： 旨在深入了解宁夏地区牛源金黄色葡萄球菌（Staphylococcus aureus，SA）所产生β-内酰胺酶的进化及结构特征并揭示其作用方式。分别以药敏试验和PCR方法检测5种β-内酰胺类药物耐药情况和耐药基因；通过生物信息学手段对检出的β-内酰胺酶BlaZ作进化分析及结构功能预测；利用分子对接和动力学模拟，分析BlaZ的作用方式。结果表明，青霉素类药物的耐药率明显高于头孢菌素；blaZ基因检出率为82.37%，并检出SA中较少报道的blaZ_TEM-1基因，检出率为26.05%；进化分析可知，BlaZ属于A型β-内酰胺酶，包含63个重要且承受进化压力的踪迹残基；分子对接发现，BlaZ易与青霉素类药物及阿维巴坦等抑制剂结合并形成稳定复合物；动力学模拟显示，BlaZ的结构柔性小于同型酶TEM-1和TEM-52，其中Ω-loop区域柔性存在差异（P<0.05）且能影响与药物的作用，即该区域柔性与结合活性呈正相关；BlaZ分别与氨苄西林和头孢噻呋结合时结构稳定性差异显著（P<0.05）。宁夏地区牛源SA产生的β-内酰胺酶BlaZ属A型，并从菌株中检出blaZ_TEM-1基因；BlaZ对青霉素类药物的结合活性高于头孢菌素且易于结合阿维巴坦等抑制剂，其中Ω-loop区域的结构柔性是BlaZ与药物结合活性的重要影响因素。

关键词: 金黄色葡萄球菌, β-内酰胺酶, 作用方式, 生物信息学, 分子模拟

Abstract: This study aimed to study the evolution and structural characteristics of β-lactamase produced by Staphylococcus aureus (SA) and its action mode in Ningxia province. Antimicrobial susceptibility test and PCR were used to detect the resistance and drug resistance genes of five β-lactams respectively; the evolution analysis and structure function prediction of β-lactamase BlaZ were made by bioinformatics; the mode of action of BlaZ was analyzed by molecular docking and dynamic simulation. The results showed that the resistance rate of Penicillins was significantly higher than Cephalosporin's; the detection rate of blaZ gene was 82.37% and the detection rate of blaZ_TEM-1 gene, which was rarely reported in SA, was 26.05%. According to evolutionary analysis, BlaZ belongs to A-type β-lactamase and contains 63 important trace residues under evolutionary pressure. Meanwhile, molecular docking showed that BlaZ is easy to combine with Penicillins, Avibatan, what's more, formed stable. The results of molecular dynamics simulation showed that the structural flexibility of BlaZ was less than that of TEM-1 and TEM-52, in which there was a difference in the flexibility of Ω-loop region (P<0.05) and it could affect the antibiotic binding. Thus, there was a positive correlation between the flexibility of the region and the binding activity; the structural stability of BlaZ was significantly different (P<0.05) when it was combined with Ampicillin and Ceftiofur, respectively. The BlaZ produced by SA in Ningxia is a A-type β-lactamase. In addition, blaZ_TEM-1 gene was detected from those strains. The bind-ing activity of BlaZ to Penicillins and Avibactam is higher than that of Cephalosporin, and the structural flexibility of Ω-loop region is an important factor affecting the binding activity.

Key words: Staphylococcus aureus, β-lactamase, action mode, bioinformatics, molecular simulation

中图分类号:

S852.611

马强, 杨蕊, 万佳宏, 常佳伟, 魏彦琴, 王桂琴. 宁夏地区牛源金黄色葡萄球菌β-内酰胺酶的分析及作用方式的研究[J]. 畜牧兽医学报, 2020, 51(5): 1138-1148.

MA Qiang, YANG Rui, WAN Jiahong, CHANG Jiawei, WEI Yanqin, WANG Guiqin. Study on β-lactamase and Its Action Mode of Staphylococcus aureus Isolated from Cows in Ningxia[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 1138-1148.

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