摩氏摩根菌噬菌体MmP1内溶素重组蛋白活性的初步鉴定
Identification of endolysin encoded by bacteriophage MmP1 against Morganella morganii
朱晓艳 1胡福泉 2谭银玲2
作者信息
- 1. 400038重庆,第三军医大学基础医学部微生物学教研室,重庆市微生物工程实验室;610083成都,成都军区总医院全军中心实验室
- 2. 400038重庆,第三军医大学基础医学部微生物学教研室,重庆市微生物工程实验室
- 折叠
摘要
目的 初步鉴定摩氏摩根菌噬菌体MmP1内溶素基因重组蛋白的生物学活性.方法 提取摩氏摩根菌细胞肽聚糖成分,采用酶谱电泳法、扩散法检测重组融合蛋白对细胞壁肽聚糖层的降解活性；采用纸片法检测重组蛋白对金黄色葡萄球的抑菌作用；测定重组蛋白与抗生素的协同抑菌作用.结果 成功提取了摩氏摩根菌细胞壁肽聚糖,酶谱法、扩散法结果表明重组蛋白对摩氏摩根菌肽聚糖具有降解作用,纸片法结果表明重组蛋白可抑制金黄色葡萄球菌的生长；当与新青霉素Ⅱ、氨苄青霉素(Amp)、羧苄青霉素(Car)3种抗生素共同作用时有协同抑菌作用,3种抗生素的MIC值分别由1.56、1.56、3.125 μg/ml降为0.39、0.78、0.195 μg/ml.结论 重组融合蛋白可降解肽聚糖、抑制金黄色葡萄球菌生长活性,且与抗生素协同作用时可降低抗生素MIC值.
Abstract
Objective To identify the biological activity of recombinant endolysin encoded by bacte-riophage MmPl against Morganella morganii. Methods The peptidoglycan of Morganella morganii was extracted. Enzymatic activity of the recombinant endolysin was detected by zymography assay and gel diffusion assay, respectively. The inhibitory activity of recombinant protein against S. Aureus culture was detected by paper disk method. The synergistic effect of recombinant protein and antibiotics was determined by MIC. Results Zymography assay and gel diffusion assay showed that Morganella morganii peptidoglycan can be hydrolyzed by the recombinant endolysin. MmPl endolysin could inhibit growth of S. Aureus. The synergistic effect was observed when MmPl endolysin was combined with antibiotics including Amp II , Amp and Car, and the MIC of Amp II , Amp and Car reduced from 0.39 μg/ml, 0.78 μg/ml and 0.195 μg/ml to 1.56 μg/ml, 1.56 μg/ml and 3. 125 μg/ml, respectively. Conclusion The recombinant endolysin can hydrolyze Morganella morganii peptidoglycan, inhibit growth of S. Aureus, and enhance bacteriostasis activity of antibiotics.
关键词
噬菌体/内溶素/摩氏摩根菌/细胞壁肽聚糖/抑菌活性Key words
bacteriophage/endolysin/Morganella morganii/peptidoglycan/antibacterial activity引用本文复制引用
出版年
2011
国家科技期刊平台
NSTL
维普
万方数据