| 猪链球菌7型噬菌体裂解酶Ly7917催化域的裂菌活性 |
| |
| 引用本文： | 吉文汇,孙亮,黄庆庆,王恒安,严亚贤,孙建和.猪链球菌7型噬菌体裂解酶Ly7917催化域的裂菌活性[J].微生物学通报,2016,43(1):156-163. |
| |
| 作者姓名： | 吉文汇 孙亮 黄庆庆 王恒安 严亚贤 孙建和 |
| |
| 作者单位： | 1. 上海交通大学农业与生物学院 上海市兽医生物技术重点实验室 上海 200240,2. 甘肃农业大学动物医学院 甘肃 兰州 730070,1. 上海交通大学农业与生物学院 上海市兽医生物技术重点实验室 上海 200240,1. 上海交通大学农业与生物学院 上海市兽医生物技术重点实验室 上海 200240,1. 上海交通大学农业与生物学院 上海市兽医生物技术重点实验室 上海 200240,1. 上海交通大学农业与生物学院 上海市兽医生物技术重点实验室 上海 200240 |
| |
| 基金项目： | 国家自然科学基金项目(No. 31172381，31372500，31272580)；国家公益性行业(农业)科研专项项目(No. 201303041)；上海市基础研究重点项目(No. 12JC1404700)；上海市科技兴农重点攻关项目(No. 2014-3-1) |
| |
| 摘 要： | 【目的】揭示链球菌噬菌体裂解酶Ly7917催化域CHAP的核心功能域,为进一步改造裂解酶提供理论依据。【方法】通过表达纯化分别从N端截短的Ly7917及从C端截短的Ly CHAP各蛋白,基于平板裂解试验和浊度递减试验,比较各截短蛋白之间的活性差异,以及添加Ca2+后各蛋白的活性变化。【结果】发现催化域蛋白Ly CHAP与Ly7917全酶的活性差异不显著,Ly CHAP的N端序列对其活性影响较大,不宜截短;而C端依次截短后,活性逐渐降低。C端截短20个氨基酸的Ly CHAP1-130,在添加1 mmol/L Ca2+后活性最强。【结论】Ly7917催化域CHAP的核心功能域为1-130 aa,推测其具有Ca2+结合区域,并发现Ly CHAP1-130在Ca2+参与下裂菌活性可媲美Ly7917全酶。预示着Ly CHAP1-130可以替代全酶应用于之后的临床试验。
|
| 关 键 词： | 猪链球菌，噬菌体裂解酶，催化结构域，裂菌活性 |
Catalytic domain of endolysin Ly7917 harbored in lysogenic phage of Streptococcus suis 7 |
| |
| JI Wen-Hui,SUN Liang,HUANG Qing-Qing,WANG Heng-An,YAN Ya-Xian and SUN Jian-He.Catalytic domain of endolysin Ly7917 harbored in lysogenic phage of Streptococcus suis 7[J].Microbiology,2016,43(1):156-163. |
| |
| Authors: | JI Wen-Hui SUN Liang HUANG Qing-Qing WANG Heng-An YAN Ya-Xian and SUN Jian-He |
| |
| Institution: | 1. Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China,2. School of Veterinary medicine, Gansu Agricultural University, Lanzhou, Gansu 730070, China,1. Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China,1. Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China,1. Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China and 1. Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China |
| |
| Abstract: | Objective] We focused on finding out the active area of catalytic domain of endolysin Ly7917 against Streptococcus suis, consequently helpful for Ly7917 modification. Methods] A group peptides truncated from N-terminal of Ly7917 or C-terminal of LyCHAP were expressed and compared with original Ly7917 and LyCHAP of the catalytic activity by plate lysis assay and turbidity decrease assay, even the situation with addition of Ca2+. Results] LyCHAP showed excellent catalytic activity as well as the full-length Ly7917. Truncated Ly7917 from N-terminal showed no catalytic activity. With the assistance of Ca2+, LyCHAP without 20 amino acids at C-terminal (LyCHAP1?130) could achieve the maximum catalytic effect even better than full-length LyCHAP. Conclusion] Ca2+-dependent LyCHAP1?130 could be the potential alternative of Ly7917 for further clinic trials. |
| |
| Keywords: | Streptococcus suis Endolysin catalytic domain bacterial lysis |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《微生物学通报》浏览原始摘要信息 |
| 点击此处可从《微生物学通报》下载免费的PDF全文 |
|
