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穿透肽嵌合金黄色葡萄球菌噬菌体裂解酶LysJS25-R9的表达及其活性分析 预览

Expression and antibacterial activity of lysin LysJS25-R9 of cell penetrating peptide chimeric S. aureus phage lysin
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摘要 前期分离得到1株具有较强裂解能力的金黄色葡萄球菌噬菌体vBSauMJS25。测序后发现,噬菌体vBSauMJS25的裂解酶基因LysJS25与裂解酶LysK同源性最高,且可以广谱裂解金黄色葡萄球菌。为提高LysJS25细胞穿透效果,设计了穿透肽嵌合裂解酶LysJS25-R9,经原核表达和纯化,体外评价了LysJS25-R9裂解活性、细胞穿透活性及细胞内杀菌活性。结果表明:LysJS25-R9对葡萄球菌表现出较好的裂解能力,杀菌效率较原裂解酶提高2.1倍。激光共聚焦及细胞内杀菌活性实验证实LysJS25-R9进入MAC-T细胞内杀菌。该研究为穿透肽嵌合裂解酶的设计提供了一个良好的例证,同时有助于裂解酶细胞穿透性改造的设计。 A strain of S. aureus phage vBSauMJS25 with good lysis ability was isolated in previous experiments. We found that the phage LysJS25 gene of vBSauMJS25 showed highly homologous with LysK, and also a broad spectrum of lysis ability against S. aureus. To improve the cell penetrating efficiency of LysJS25, we designed a cell penetrating peptide chimeric S. aureus phage lysin. To explore the lysis activity of chimeric lysin, LysJS25 was expressed in prokaryotic cells and purified. The lysis activity and the cell penetrating activity of LysJS25-R9 were determined. Our results showed that the lysis activity against S. aureus of LysJS25-R9 was 2.1 times higher than LysJS25. The laser confocal and intracellular lysis activity assay also showed that LysJS25-R9 could penetrate bovine mammary epithelial cells to kill intracellular S. aureus. These results not only provide strong evidence for the study cell penetrating peptide chimeric phage lysin, but also contribute to the design of cell penetrating peptide chimeric lysin.
作者 孙利厂 周艳 张莉莉 庞茂达 何涛 包红朵 张辉 王冉 Sun Li-chang, Zhou Yan, Zhang Li-li, Pang Mao-da, He Tao, Bao Hong-duo, Zhang Hui, Wang Ran (Jiangsu Academy of Agricultural Sciences Institute of Food Safety and Nutrition, Key Lab of Food Quality and Safety of Jiangsu Province--State Key Laboratory Breeding Base, Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agricultrue, Key Laboratory of Animal-derived Food safety of Jiangsu Province/Jiangsu Academy of Agricultural Sciences, Nanjing 210014)
出处 《中国抗生素杂志》 CSCD 北大核心 2017年第10期865-870,共6页 Chinese Journal of Antibiotics
基金 江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地自主研究课题(No.3201614) 江苏省基础研究计划(自然科学基金)项目(No.BK20160585) 江苏省农业科技自主创新项目(No.CX(16)1060)
关键词 金黄色葡萄球菌 噬菌体 裂解酶 细胞穿透肽 S. aureus Phage Lysin Cell penetrating peptide
作者简介 孙利厂,男,生于1985年,助理研究员,博士,主要从事食源性病原微生物防控研究,E-mail:sunlichang1860@163.com 通讯作者,E-mail:wangran@jaas.ac.cn
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