食品科学 ›› 2017, Vol. 38 ›› Issue (8): 74-79.doi: 10.7506/spkx1002-6630-201708013

• 生物工程 • 上一篇    下一篇

南海海域动物肠道中抗菌活性乳酸菌的分离及多样性分析

焦冬冬,任文彬,赵鸭美,侯小琴,刘 颖,孙力军,王雅玲   

  1. 1.广东海洋大学食品科技学院,广东省水产品加工与安全重点实验室,水产品深加工广东普通高等学校重点实验室, 广东 湛江 524088;2.仲恺农业工程学院轻工食品学院,广东 广州 510230;3.湖州环境科技创新中心,浙江 湖州 313000
  • 出版日期:2017-04-25 发布日期:2017-04-24
  • 基金资助:
    广东省科技计划项目(2014A020217018;2016A020222014); 广东省水产品加工与安全重点实验室开放基金课题(GDPKLAPPS1503);国家自然科学基金面上项目(31371777;31171634)

Separation and Diversity of Lactic Acid Bacteria Possessing Antimicrobial Activity Isolated from the Intestine of Marine Animals in South China Sea

JIAO Dongdong, REN Wenbin, ZHAO Yamei, HOU Xiaoqin, LIU Ying, SUN Lijun, WANG Yaling   

  1. 1. Key Laboratory of Aquatic Product Processing and Safety of Guangdong, Key Laboratory of Advanced Processing of Aquatic Products of Guangdong Higher Education Institution, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, China; 2. College of Food Science and Technology, Zhongkai University of Agriculture and Engineering, Guangzhou 510230, China; 3. Huzhou Environmental Science and Technology Innovation Center, Huzhou 313000, China
  • Online:2017-04-25 Published:2017-04-24

摘要: 以湛江南海海域常见的10 种海洋动物为研究对象,采用倾注平板法分离肠道乳酸菌,以单核细胞增生李斯特氏菌(Listeria monocytogenes)等5 株有害菌为指示菌,采用双层牛津杯扩散法测定其抗菌活性,通过建立16S rRNA基因序列的系统发育树对具有抗菌活性的乳酸菌进行多样性分析。结果表明:根据形态观察和部分生理生化实验去重复后,共分离出43 株有溶钙圈、革兰氏染色呈阳性、过氧化氢酶反应为阴性的乳酸菌,并且均不同程度地具有抗菌活性。对其中的13 株抗菌能力强、抗菌谱广的乳酸菌进行16S rRNA分子生物学鉴定与分析,发现它们分属于厚壁菌门(Firmicutes)的4 个科6 个属,且除菌株LY-87与模式菌株Lactococcus lactis subsp. lactis的16S rRNA基因序列相似性为100%外,其他菌株与其系统发育关系最密切的典型菌株之间都存在着一定的遗传差异(相似性为95.7%~99.9%),其中菌株PQ-26与模式菌株Pediococcus pentosaceus的相似性仅为95.7%,可能是Pediococcus属中潜在的新种。研究表明湛江南海海域动物肠道中存在较为丰富的抗菌活性乳酸菌,并可能蕴藏着乳酸菌新种,其抗菌物质资源有待进一步研究与开发。

关键词: 海洋动物肠道, 抗菌活性, 乳酸菌, 16S rRNA基因, 多样性

Abstract: Lactic acid bacteria (LAB) were isolated from the intestine of 10 common kinds of marine animals collected from the South China Sea by the pour plate method. The antimicrobial activity of the LAB isolates was determined by Oxford cup diffusion method against five pathogenic indicator strains including E. coli, V. parahaemolytics, S. aureus, B. subtilis, and L. monocytogenes. The diversity analysis of these LAB strains was conducted by establishing phylogenetic tree based on 16S rRNA gene sequences. The results showed that 43 isolates, which presented soluble calcium circle and were positive by Gram staining and negative in catalase reaction, were identified as LAB according to morphological observation and partial physiological and biochemical experiments, and they all possessed antimicrobial activity in different degrees. Out of these, 13 strains were found to have stronger bacteriostatic ability and a wider antimicrobial spectrum, and they were further identified based on phyogenetical analysis of 16S rRNA sequence. The results indicated that they belonged to 6 genera in 4 families of Firmicutes. Although strain LY-87 and the type strain of Lactococcus lactis subsp. lactis shared 100% 16S rRNA gene sequence similarity, there were genetic differences between other LAB strains and the typical strains showing the closest phylogenetic relationship with them (similarity ranging from 95.7% to 99.9%). The similarity between strain PQ- 26 and the type strain of Pediococcus pentosaceus was only 95.7% suggesting that it is likely to be a potential new species of Pediococcus. This research indicates that LAB with antimicrobial activity are diverse and there may exist potential new species in the intestine of animals from the South China Sea in Zhanjiang. The antimicrobial substances produced by these bacteria need to be further studied and developed.

Key words: marine animal intestine, antimicrobial activity, lactic acid bacteria, 16S rRNA gene, diversity

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