武汉轻工大学学报2024,Vol.43Issue(5) :10-17.DOI:10.3969/j.issn.2095-7386.2024.05.002

副溶血弧菌噬菌体裂解酶LysF23s2和LysH256D1的表达及活性分析

Characterisation of Vibrio parahaemolyticus phage lyase LysF23s2 and LysH256D1

石甜 史聪聪 衡冰冰 夏海 周敏
武汉轻工大学学报2024,Vol.43Issue(5) :10-17.DOI:10.3969/j.issn.2095-7386.2024.05.002

副溶血弧菌噬菌体裂解酶LysF23s2和LysH256D1的表达及活性分析

Characterisation of Vibrio parahaemolyticus phage lyase LysF23s2 and LysH256D1

石甜 1史聪聪 1衡冰冰 1夏海 1周敏1
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作者信息

  • 1. 武汉轻工大学食品科学与工程学院,武汉 430023
  • 折叠

摘要

由于副溶血弧菌对食品安全构成了巨大的威胁,抗生素的滥用导致耐药问题日趋严重.噬菌体及其裂解酶能够迅速、高效杀灭细菌,不易导致细菌耐药性,成为可替代抗生素的新型抑菌剂.该文将两种副溶血弧菌噬菌体裂解酶通过BL21和T7表达体系在16 ℃、37 ℃下进行表达,成功表达并纯化裂解酶后,分析了裂解酶对副溶血弧菌的裂解能力、酶的热稳定性以及金属离子对其活性的影响.结果表明,LysF23s2和LysH256D1在BL21中低温诱导时,有更高浓度的表达.LysF23s2和LysH256D1在浓度为10 μmol/L时对宿主细菌的裂解率分别达到了75.24%和71.00%.在90 ℃处理30 min后,LysF23s2和LysH256D1的裂解率降至10.00%和 8.82%,仍保持了一定活性.Na2+、Ca2+和 Zn2+对 LysF23s2 和 LysH256D1 的活性有一定抑制作用.该研究通过大肠杆菌体系表达能够裂解副溶血弧菌的噬菌体裂解酶LysF23s2和LysH256D1,得到了两种具有较好的裂解活性和耐热性的裂解酶.

Abstract

Vibrio parahaemolyticus poses a great threat to food safety,while the misuse of antibiotics has led to the increase of drug-resistant bacteria.In this paper,two Vibrio parahaemolyticus phage lytic enzymes were expressed using the BL21 and T7 expression systems at 16 ℃ and 37 ℃.After successful expression and purification of the lyase enzyme,the lyase enzyme was analyzed for its lysing ability and thermal stability against Vibrio parahaemolytic-us,as well as the effect of metal ions on its activity.The results showed that LysF23s2 and LysH256D1 were ex-pressed at higher concentrations when induced by low temperature in BL21.The lysis rates of LysF23s2 and Ly-sH256D1 reached 75.24%and 71%,respectively,against the host bacteria at a concentration of 10 μmol/L.The lysis rates of LysF23s2 and LysH256D1 were reduced to 10%and 8.82%after being incubated at 90 ℃ for 30 min,but still maintained some activity.The activity of LysF23s2 and LysH256D1 was inhibited by Na2+,Ca2+and Zn2+,In this study,the phage lytic enzymes LysF23s2 and LysH256D1,which are capable of lysing Vibrio parahae-molyticus,were expressed in the E.coli system,and two lytic enzymes with good lysing activity and heat resistance were obtained.

关键词

副溶血弧菌/噬菌体裂解酶/酶学表征

Key words

Vibrio parahaemolyticus/phage lyase/enzymatic characterization

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基金项目

湖北省教育厅科学技术研究项目(D20221611)

湖北省教育厅科学技术研究项目(Q20211606)

武汉轻工大学校级科研项目(2021Y02)

出版年

2024
武汉轻工大学学报
武汉工业学院

武汉轻工大学学报

影响因子:0.356
ISSN:1009-4881
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