| 摘要: |
| 试验利用溶钙圈法及牛津杯双层平板法,从国内外优良发酵肉品中分离筛选产细菌素的乳酸菌,通过形态学、生理生化学鉴定,16S rDNA及种间特异性基因PCR扩增将菌株鉴定到种,并对其细菌素相关基因进行分析,初步确定细菌素种类。结果表明:在排除有机酸、过氧化氢等干扰因素后,从分离纯化到的92株乳酸菌中筛选到一株对指示菌仍有明显抑制作用的菌株L-ZS9,该菌株所产抑菌物质对蛋白酶(酸性蛋白酶、蛋白酶K、胰蛋白酶、胃蛋白酶和中性蛋白酶)敏感,而α-淀粉酶对其活性基本无影响,因而确定L-ZS9为细菌素产生菌。经鉴定L-ZS9为类植物乳杆菌,进一步对菌株细菌素相关基因进行PCR扩增表明,L-ZS9含有Ⅱb类细菌素Plantaricin EF,Plantaricin JK,Ⅱc类细菌素Plantaricin A及Plantaricin N基因编码序列,此外plnE、plnF、plnJ和 plnK的序列分析结果显示,plnF和plnK的成熟肽序列与Lactobacillus plantarum C11(X94434)相应的细菌素成熟肽序列完全相同,plnE和plnJ的成熟肽序列仅出现一处氨基酸突变,因而确定类植物乳杆菌L-ZS9为天然的多种细菌素产生菌。 |
| 关键词: 乳酸菌 筛选 细菌素 类植物乳杆菌 |
| DOI：10.11841/j.issn.1007-4333.2013.04.26 |
| 投稿时间：2013-01-15 |
| 基金项目:国家自然科学基金项目(31071591); 产业技术体系北京市创新团队项目(SCGWZJ 20121105-2) |
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| Screening of bacteriocin producing lactic acid bacteria and the analysis of bacteriocin related genes |
|
ZHANG Xu, ZHAO Bin, ZHANG Xiang-mei, LI Ping-lan
|
| (College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China) |
| Abstract: |
| In order to obtain a bacteriocin producing strain,lactic acid bacteria were separated and screened with dissolve calcium circle and oxford cup double-plate method from premium fermentation meat.Morphological,physiological biochemistry method,16S rDNA and specific gene PCR amplification were used to identify the strain to species,and the bacteriocin related gene was analyzed to determine bacteriocin types.The results showed that,after eliminating organic acid,hydrogen peroxide and other interference factors,L-ZS9 still had antibacterial activity among the 92 strains and its antibacterial substances were sensitive to protease (acid protease,protease K,trypsin,pepsin,neutral protease),while insensitive to alpha amylase.Therefore,L-ZS9 was recognized as bacteriocin producing bacteria and identified as Lactobacillus paraplantarum.Further bacteriocin related gene PCR amplification showed that L-ZS9 contained Ⅱb plantaricins EF,JK,Ⅱc plantaricin A and plantaricin N encoding gene.The mature peptide sequences of plnF,plnK were identical with Lactobacillus plantarum C11 (X94434) and only one amino acid mutation occurred in plnE,plnJ mature peptide sequences.Therefore,the Lactobacillus paraplantarum L-ZS9 was a nature multi-bacteriocinogenic strain. |
| Key words: lactic acid bacteria screening bacteriocin Lactobacillus paraplantarum |
