乳酸菌细菌素plantaricin JK的异源表达、分离纯化及抗菌活性

doi:10.3969/j.issn.1004-1524.2017.02.21

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (2): 332-337.DOI: 10.3969/j.issn.1004-1524.2017.02.21

乳酸菌细菌素plantaricin JK的异源表达、分离纯化及抗菌活性

蒋晗^{1, 2}, 刘娇¹, 郦萍¹, 顾青^{1, *}

1.浙江工商大学浙江省食品微生物技术研究重点实验室,浙江杭州 310018;
2.中国计量大学浙江省海洋食品品质及危害物控制技术重点实验室,浙江杭州 310018

收稿日期:2016-12-12 出版日期:2017-02-15 发布日期:2017-03-06
通讯作者: 顾青,E-mail:guqing2002@hotmail.com
作者简介:蒋晗(1987—),女,浙江杭州人,博士研究生,研究方向为食品生物技术。E-mail:jianghan825@126.com
基金资助:
国家国际科技合作专项(2013DFA32330); 国家自然科学基金项目(31540044,31271821); 国家高技术研究发展计划(863计划) (2014AA022210-08)

Heterologous expression and purification of plantaricin JK and analysis of its antibacterial activity

JIANG Han^{1, 2}, LIU Jiao¹, LI Ping¹, GU Qing^{1, *}

1. Key Laboratory for Food Microbial Technology of Zhejiang Province, Zhejiang Gongshang University, Hangzhou 310018, China;
2. Key Laboratory of Marine Food Quality and Hazard Controlling Technology of Zhejiang Province, China Jiliang University, Hangzhou 310018, China

Received:2016-12-12 Online:2017-02-15 Published:2017-03-06

摘要/Abstract

摘要： 为获得高产双肽细菌素plantaricin JK,通过PCR扩增获得pln J和pln K基因,构建表达载体pET32a(+)-pln J-BL21(DE3)和pET32a(+)-pln K-BL21(DE3),经IPTG诱导表达后,利用镍亲和层析柱进行分离纯化,重组蛋白经肠激酶酶切纯化后,细菌素pln J和pln K的产量分别为2~3、5~8 mg·L^-1。利用牛津杯平板抑菌法研究抗菌活性,结果显示细菌素pln J和pln K对柠檬色葡萄球菌(Staphylococcus citreus)、大肠埃希菌(Escherichia coli)、藤黄微球菌(Micrococcus luteus)和单核细胞增生李斯特菌(Listeria monocytogenes)都有明显抑制作用,且双肽细菌素plantaricin JK的抑菌圈直径大于单独的细菌素pln J或pln K。

关键词: plantaricin JK, 异源表达, 分离纯化, 抗菌活性

Abstract: In order to obtain the high-yield two-peptide bacteriocin plantaricin JK, pln J and pln K were successfully heterologously expressed in Escherichia coli BL21 (DE3), and were induced by IPTG. Then the two peptides were expressed as His₆-tag fusion proteins and were separated by Ni²⁺ chelating affinity chromatography. The fusion proteins were cleaved by enterokinase and further purified. The yields of pln J and pln K were around 2-3 and 5-8 mg·L^-1. The antibacterial activity of the peptides against 4 indicator strains, i.e. Staphylococcus citreus, Escherichia coli, Micrococcus luteus, Listeria monocytogenes, was analyzed by agar diffusion method. It was shown that pln J and pln K both could inhibit the growth of the 4 indicator strains, but plantaricin JK had larger inhibition zone than pln J or pln K alone.

Key words: plantaricin JK, heterologous expression, expression and purification, antibacterial activity

中图分类号:

Q786

蒋晗, 刘娇, 郦萍, 顾青. 乳酸菌细菌素plantaricin JK的异源表达、分离纯化及抗菌活性[J]. 浙江农业学报, 2017, 29(2): 332-337.

JIANG Han, LIU Jiao, LI Ping, GU Qing. Heterologous expression and purification of plantaricin JK and analysis of its antibacterial activity[J]. , 2017, 29(2): 332-337.

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乳酸菌细菌素plantaricin JK的异源表达、分离纯化及抗菌活性

Heterologous expression and purification of plantaricin JK and analysis of its antibacterial activity

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