噬菌体裂解酶结构域重构及其裂解功能分析

doi:10.7668/hbnxb.2017.06.020

华北农学报 ›› 2017, Vol. 32 ›› Issue (6): 134-138. doi: 10.7668/hbnxb.2017.06.020

噬菌体裂解酶结构域重构及其裂解功能分析

张辉¹, 周艳¹, 包红朵¹, 杨振泉³, 朱如意³, 周晨露³, 张莉莉¹, 王冉^1,2

1. 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, 江苏省农业科学院, 江苏南京 210014;
2. 江苏省肉类生产与加工质量安全控制协同创新中心, 江苏南京 210095;
3. 扬州大学食品科学与工程学院, 江苏扬州 225009

收稿日期:2017-08-09 出版日期:2017-12-28
通讯作者: 王冉(1973-),女,河北石家庄人,研究员,博士,主要从事食品污染物监测及防控研究。
作者简介:张辉(1978-),女,新疆吐鲁番人,副研究员,博士,主要从事食源性病原菌监测及生物防控研究。
基金资助:
国家自然科学基金项目（31671955）；江苏省农业科技自主创新资金项目（cx（13）3086）；江苏省自然科学基金项目（BK20161373）

Domain Reconstruction and Lysis Activity of Phage Lysin

ZHANG Hui¹, ZHOU Yan¹, BAO Hongduo¹, YANG Zhenquan³, ZHU Ruyi³, ZHOU Chenlu³, ZHANG Lili¹, WANG Ran^1,2

1. Jiangsu Key Laboratory of Food Quality and Safety-State Key Laboratory Cultivation Base of MOST, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;
2. Jiangsu Collaborative Innovation Center of Meat Production and Processing, Quality and Safety Control, Nanjing 210095, China;
3. College of Food Science and Engineering, Yangzhou University, Yangzhou 225009, China

Received:2017-08-09 Published:2017-12-28

摘要/Abstract

摘要： 为了获得广谱噬菌体裂解酶，根据裂解酶结构及种属特异性，将金黄色葡萄球菌噬菌体裂解酶Ply187 CHAP及酶K的SH3b重新合成，并同时将末端引入单核细胞增生性李斯特菌噬菌体裂解酶LysZ5结合结构域，构建具有能够裂解金黄色葡萄球菌及李斯特菌的多功能裂解酶。SDS-PAGE结果表明，融合结构域的重构酶蛋白Ply187KS-Z5大小约为58 kDa，其在金黄色葡萄球菌及李斯特菌平板表面能够形成透明裂解圈。分析对金黄色葡萄球菌及李斯特菌在生肉表面的裂解活性表明，嵌合酶蛋白Ply187-KS能够有效裂解金黄色葡萄球菌，作用1 h后菌落数降至2.16 log，而重构酶Ply187KS-Z5不仅裂解金黄色葡萄球菌并同时能够裂解李斯特菌，且重构酶针对金黄色葡萄球菌噬菌体的裂解活性与Ply187-KS裂解效果相当，从而表明裂解酶结合结构域重组将是获得高效广谱裂解酶的新途径。

关键词: 裂解酶, 金黄色葡萄球菌, 单核细胞增生性李斯特菌

Abstract: In order to get wide host-range phage lysin,the cell-binding domains of Staphylococcus aureus Ply187 CHAP and SH3b (lysin K) were synthesized based on the sequence,according to the specificity and structure of the phage lysin.The cell-binding domain was amplified from Listeria monocytogenes phage lysin LysZ5.All the domain were reconstructed and the lysis function were analyzed.From the SDS-PAGE,reconstruction lysin Ply187KS-Z5,containing two cell-binding domains,showed the specific protein band at 58 kDa.Clear zone observed after Lysin Ply187KS-Z5 added on the plate of both S.aureus and L.monocytogenes. Lysin Ply187-KS could reduce number of S. aureus to 2.16 log in 1 hour,but the reconstruction lysin Ply187KS-Z5 was able to inhibit the growth of both S.aureus and L.monocytogenes.Therefore,domain reconstruction should be a new approach to develop wide-host-range lysin.

Key words: Lysin, Staphylococcus aureus, Listeria monocytogenes

中图分类号:

S432.4

张辉, 周艳, 包红朵, 杨振泉, 朱如意, 周晨露, 张莉莉, 王冉. 噬菌体裂解酶结构域重构及其裂解功能分析[J]. 华北农学报, 2017, 32(6): 134-138. doi: 10.7668/hbnxb.2017.06.020.

ZHANG Hui, ZHOU Yan, BAO Hongduo, YANG Zhenquan, ZHU Ruyi, ZHOU Chenlu, ZHANG Lili, WANG Ran. Domain Reconstruction and Lysis Activity of Phage Lysin[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(6): 134-138. doi: 10.7668/hbnxb.2017.06.020.

http://www.hbnxb.net/CN/Y2017/V32/I6/134

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