食品科学 ›› 2018, Vol. 39 ›› Issue (18): 145-151.doi: 10.7506/spkx1002-6630-201818023

• 生物工程 • 上一篇    下一篇

1 株水开菲尔粒中植物乳杆菌的鉴定及产细菌素基因分析

梁新红1,冉军舰1,*,孙华迪2,赵瑞香1,焦凌霞1,卢艳清1   

  1. (1.河南科技学院食品学院,河南省高校重点实验室培育基地,新乡市农产品加工工程技术研究中心,河南?新乡 453003;2.河南科技学院新科学院,河南?新乡 453003)
  • 出版日期:2018-09-25 发布日期:2018-09-18
  • 基金资助:
    河南省科技计划项目(182102110109);河南省科技厅科技攻关项目(182102410098); 河南省高校重点科研项目(18A550006 );河南省高校科技创新人才支持计划项目(17HASTIT037)

Identification of a Lactobacillus plantarum Strain Isolated from Water Kefir Grains and Sequence Analysis of Its Bacteriocin Genes

LIANG Xinhong1, RAN Junjian1,*, SUN Huadi2, ZHAO Ruixiang1, JIAO Lingxia1, LU Yanqing1   

  1. (1. Xinxiang Engineering Technology Research Center for Agricultural Products Processing, Key Lab of Breeding Base of College ofHenan Province, School of Food Science, Henan Institute of Science and Technology, Xinxiang 453003, China; 2. Xinke College, Henan Institute of Science and Technology, Xinxiang 453003, China)
  • Online:2018-09-25 Published:2018-09-18

摘要: 目的:对从水开菲尔粒中分离得到1 株能产生抑菌物质的菌株QF01进行菌种鉴定,并对其产细菌素进行实验、鉴定和基因序列分析。方法:通过牛津杯法测定抑菌能力,采用聚合酶链式反应(polymerase chain reaction,PCR)扩增16S rDNA和细菌素相关基因并测序,利用ProParam tool、TMHMM 2.0、InterProScan、SOPM和SWISS-MODEL在线软件对基因编码产物进行分析。结果:该菌株产的细菌素对大肠杆菌(Escherichia coli JM109)有明显的抑制作用,通过16S rDNA序列分析初步确定该菌株属于植物乳杆菌(Lactobacillus plantarum)。该菌株含有plnD、plnEF、plnV、plnR四种基因,其中plnV基因编码产物有跨膜螺旋结构,其结构存在信号肽特征。此外,从三级结构的模型预测得到4?种基因的编码产物均存在α-螺旋、β-转角、伸展链和无规则卷曲。结论:从水开菲尔粒分离得到的L. plantarum QF01菌株,含有plnD、plnEF、plnV、plnR细菌素基因,对大肠杆菌有很好的抑制作用。

关键词: 植物乳杆菌, 细菌素, 抑菌活性, 基因克隆, 生物信息学

Abstract: Objective: The bacteriocin-producing strain QF01, isolated from water kefir grains, was identified and the antibacterial activity of bacteriocin from the isolate was evaluated. The gene sequences encoding bacteriocin were analyzed as well. Methods: The antibacterial activity of bacteriocin was determined by the Oxford cup method. The bacteriocin-related genes were amplified by PCR, 16S rDNA analysis was used for bacterial identification and bacteriocin gene analysis was carried out by using online software (ProParam tool, TMHMM 2.0, InterProScan, SOPM and SWISS-MODEL). Results: Bacteriocin produced by strain QF01 demonstrated a strong inhibitory activity against Escherichia coli JM109. This strain was identified as Lactobacillus plantarum based on its 16S rDNA sequence. The four bacteriocin-encoding genes plnD, plnEF, plnV and plnR were found in Lactobacillus plantarum QF01. The plnV gene encoding product had a helical transmembrane structure with signal peptide characteristics at its N-terminal. In addition, each of the four gene encoding products contained α-helix, β-turn, extended chain and random coil structures, as predicted from the tertiary structure model. Conclusion: L. plantarum QF01 contains the bacteriocin-encoding genes plnD, plnEF, plnV and plnR, and has a strong inhibitory activity against E. coli.

Key words: Lactobacillus plantarum, bacteriocin, antibacterial activity, gene cloning, bioinformatics

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