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食品与发酵工业  2021, Vol. 47 Issue (9): 107-113    DOI: 10.13995/j.cnki.11-1802/ts.026139
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
大肠杆菌O157∶H7噬菌体EC-p9的内溶酶和穿孔素的特性预测及克隆表达
解天慧, 石慧*
(西南大学 食品科学学院,重庆,400715)
Characteristics prediction and expression of endolysin and holin from E.coli O157∶H7 phage EC-p9
XIE Tianhui, SHI Hui*
(College of Food Science,Southwest University,Chongqing 400715,China)
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摘要 大肠杆菌O157∶H7是一种常见的能严重威胁公共卫生安全的食源性致病菌,噬菌体及其内溶酶、穿孔素是新型的抑菌剂。该文利用十聚体寡核苷酸随机引物初步鉴定了噬菌体EC-p9;应用 PCR技术获得该噬菌体的内溶酶Lys 9和穿孔素Hol 9的基因;利用生物学信息预测了这2种蛋白的特性;通过大肠杆菌表达体系表达了这2种蛋白。结果表明,噬菌体EC-p9是dsDNA肌尾科噬菌体,具有“穿孔素-内溶酶”裂解体系。内溶酶Lys 9是一种不含跨膜结构的肽聚糖水解酶。穿孔素Hol 9是Ⅲ型穿孔素,仅有1个跨膜结构。内溶酶Lys 9和穿孔素Hol 9都能通过大肠杆菌表达体系成功表达,但穿孔素Hol 9的表达会对大肠杆菌BL21(DE3)产生毒害作用,导致表达量较少。最终获得了质量浓度为2.49 g/L的内溶酶Lys 9和0.95 g/L的穿孔素Hol 9。该研究为开展内溶酶Lys 9和穿孔素Hol 9的抑菌能力以及噬菌体EC-p9的裂解体系的研究奠定了基础。
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解天慧
石慧
关键词:  大肠杆菌O157∶H7  噬菌体EC-p9  内溶酶Lys9  穿孔素Hol9    
Abstract: Escherichia coli O157∶H7 is a common foodborne pathogen that severely threatens public health. Phage, endolysin and holin are new bacteriostatic agents. In this study, phage EC-p9 was identified by random primer for decammeric oligonucleotide, genes of endolysin Lys 9 and holin Hol 9 of the phage were obtained through PCR, properties of the two proteins were predicted through in-silico analysis, and the two proteins were expressed by E.coli. The results indicated that phage EC-p9 was dsDNA phage with a “holin-endolysin” lysis system. Endolysin Lys 9 was a peptidoglycan hydrolase without a transmembrane structure. Holin Hol 9 was a type III holin with only one transmembrane structure. Endolysin Lys 9 and holin Hol 9 were successfully expressed in E.coli BL21 (DE3), with yield of 2.49 and 0.95 g/L, respectively. Only limited amount of holin Hol 9 was obtained due to its toxic effect on E.coli. The results laid a foundation for research of the bacteriostatic ability of endolysin Lys 9 and holin Hol 9, and also for investigation of the lysis system of phage EC-p9.
Key words:  E.coli O157∶H7    phage EC-p9    endolysin Lys 9    holin Hol 9
收稿日期:  2020-11-10      修回日期:  2020-12-01           出版日期:  2021-05-15      发布日期:  2021-06-03      期的出版日期:  2021-05-15
基金资助: 国家自然科学基金面上项目(32072322);重庆市技术创新与应用发展项目面上项目(cstc2019jscx-msxmX0348);重庆市留学人员创业创新计划优秀项目(cx2019017);中央高校基本科研业务费重点项目(XDJK2020B043)
作者简介:  硕士研究生(石慧副教授为通讯作者,E-mail:shi_hui_1986@163.com)
引用本文:    
解天慧,石慧. 大肠杆菌O157∶H7噬菌体EC-p9的内溶酶和穿孔素的特性预测及克隆表达[J]. 食品与发酵工业, 2021, 47(9): 107-113.
XIE Tianhui,SHI Hui. Characteristics prediction and expression of endolysin and holin from E.coli O157∶H7 phage EC-p9[J]. Food and Fermentation Industries, 2021, 47(9): 107-113.
链接本文:  
http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.026139  或          http://sf1970.cnif.cn/CN/Y2021/V47/I9/107
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