植物乳杆菌KLDS1.0391所产细菌素的纯化

食品科学 ›› 2013, Vol. 34 ›› Issue (3): 59-63.

植物乳杆菌KLDS1.0391所产细菌素的纯化

王辉¹,孟祥晨²

1. 东北农业大学乳品科学教育部重点实验室
2. 东北农业大学食品学院

收稿日期:2011-10-20 修回日期:2013-01-07 出版日期:2013-02-15 发布日期:2017-12-29
通讯作者: 孟祥晨 E-mail:xchmeng@163.com
基金资助:
黑龙江省留学归国科学基金项目

Purification of the bacteriocin produced by Lactobacillus plantarum KLDS1.0391

Received:2011-10-20 Revised:2013-01-07 Online:2013-02-15 Published:2017-12-29
Contact: [中]孟祥晨 [英]MENG Xiang-Chen E-mail:xchmeng@163.com

摘要/Abstract

摘要： 目的是为了分离纯化植物乳杆菌KLDS1.0391代谢产生的细菌素。采用蛋白质层析技术分离细菌素，采用质谱测定纯化后细菌素的分子量。经过实验比对，最终确定的分离纯化路线为：植物乳杆菌KLDS1.0391的无细胞上清液首先经过饱和度为70%的硫酸铵沉淀，收集的沉淀溶于1/40体积0.02 mol/L的乙酸钠缓冲液(pH 6.5)，复溶后的样品采用SP Sepharose fast flow阳离子填料和COLUMN XK 16/20层析柱进行阳离子纯化，之后采用SOURCE 15RPC ST 4.6/150进行反相纯化。对收集到的具有抑菌活性的样品经高效液相色谱检测，判断其纯度达到质谱分析要求，经Tricine-SDS-PAGE检测，初步判定其分子量约为2000 kDa。采用质谱MALDI-TOF测定该细菌素的分子量为1770 kDa。

Abstract: The purpose of this study is to purify the bacteriocin produced by Lb. plantarum KLDS1.0391. The bacteriocin was separated by chromatography techniques and the molecular mass of purified bacteriocin was determined by mass spectrometric analysis. The optimal method of purification was finally determined as follow: Cell-free supernatants of Lb. plantarum KLDS1.0391 was precipitated by 70% ammonium sulphate. Then the precipitates were dissolved in 1/40 volume of 0.05 M sodium acetate (pH 6.5), followed by cation exchange chromatography (using COLUMN XK 16/20 chromatographic column filled with SP Sepharose fast flow), and reverse-phase chromatography (using SOURCE 15RPC ST 4.6/100 further purify the solution for two times). The purity of the purified samples was determined by high-performance liquid chromatography (HPLC), and the molecular mass of samples was estimated on Tricine-SDS-PAGE, which concluded that the molecular mass was approximately 2000 kDa. Matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) MS analysis of purified bacteriocin revealed that the molecular mass was approximately 1770 kDa.

中图分类号:

TS252.1

王辉孟祥晨. 植物乳杆菌KLDS1.0391所产细菌素的纯化[J]. 食品科学, 2013, 34(3): 59-63.

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