[1]蔡幸哲,彭林,刘珂芮,等.沙门氏菌噬菌体裂解酶LysLorf22的制备及溶菌活性分析[J].江苏农业学报,2020,(01):212-218.[doi:doi:10.3969/j.issn.1000-4440.2020.01.030]
　CAI Xing-zhe,PENG Lin,LIU Ke-rui,et al.Preparation and activity analysis of endolysin LysLorf22 derived from Salmonella bacteriophage[J].,2020,(01):212-218.[doi:doi:10.3969/j.issn.1000-4440.2020.01.030]
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沙门氏菌噬菌体裂解酶LysLorf22的制备及溶菌活性分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2020年01期
页码:
212-218
栏目:
加工贮藏·质量安全
出版日期:
2020-02-29

文章信息/Info

Title:
Preparation and activity analysis of endolysin LysLorf22 derived from Salmonella bacteriophage
作者:
蔡幸哲; 彭林; 刘珂芮; 刘书亮; 李诚; 刘爱平
（四川农业大学食品学院，四川雅安625014）
Author(s):
CAI Xing-zhe; PENG Lin; LIU Ke-rui; LIU Shu-liang; LI Cheng; LIU Ai-ping
(College of Food Science, Sichuan Agricultural University, Ya′an 625014, China)
关键词:
沙门氏菌; 噬菌体; 裂解酶; 外膜渗透剂
Keywords:
Salmonella; bacteriophage; endolysin; outer membrane permeabilizing agent
分类号:
TS207.7
DOI:
doi:10.3969/j.issn.1000-4440.2020.01.030
文献标志码:
A
摘要:
根据沙门氏菌噬菌体Lumpael的全基因组序列，预测该噬菌体裂解酶基因并对其进行生物信息学分析；优化其在大肠杆菌表达系统中的表达，并分析噬菌体裂解酶LysLorf22与外膜渗透剂最优组合的溶菌效果。结果表明，裂解酶LysLorf22的相对分子质量为1.76×104，等电点为9.14，其较优表达条件为：表达菌株Escherichia coli BL21，37 ℃诱导4 h。LysLorf22裂解谱较宽，最佳工作浓度为375 nmol/L，在30 min内可使氯仿处理的Salmonella Enteritidis ATCC 13076菌悬液OD600下降0.72；375 nmol/L的LysLorf22与0.5 mmol/L乙二胺四乙酸二钠联用对Salmonella Enteritidis ATCC 13076溶菌效果最佳，在2 h内可使活菌数从1.62×109CFU/ml下降至4.31×108CFU/ml。
Abstract:
In this study, the gene encoding endolysin was predicted and analyzed by bioinformatic software based on the genomic sequence of Salmonella bacteriophage Lumpael. Besides, the expression of LysLorf22 in Escherichia coli expression system was optimized, and the bactericidal effect of LysLorf22 combined with outer membrane permeabilizing agents was analyzed. The results showed that the molecular weight of LysLorf22 was 1.76×104, and the isoelectric point was 9.14. The optimal expression condition of LysLorf22 was using expression strain Escherichia coli BL21, induced at 37 ℃ for four hours. LysLorf22 had a broad lytic spectrum, and its optimal working concentration was 375 nmol/L, in which the OD600 of chloroform-treated Salmonella Enteritidis ATCC 13076 could be reduced by 0.72 in 30 minutes. The combination of 375 nmol/L LysLorf22 and 0.5 mmol/L disodium ethylenediamine tetraacetate showed the best lytic effect on Salmonella Enteritidis ATCC 13076, which reduced the viable cells from 1.62×109CFU/ml to 4.31×108CFU/ml in two hours.

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备注/Memo

备注/Memo:
收稿日期：2019-06-22基金项目：国家自然科学基金项目（31801629）；四川农业大学大学生创新训练计划项目（201910626138）作者简介：蔡幸哲（1998-），女，浙江乐清人，本科生，主要从事食品微生物研究。(E-mail)908301008@qq.com通讯作者：刘爱平，(E-mail)aipliu@outlook.com
更新日期/Last Update: 2020-03-13