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中国精品科技期刊2020
刘英丽, 丁立, 谢良需, 王静, 张慧娟. 产细菌素乳酸菌的筛选与鉴定[J]. 食品工业科技, 2017, (17): 146-151. DOI: 10.13386/j.issn1002-0306.2017.17.028
引用本文: 刘英丽, 丁立, 谢良需, 王静, 张慧娟. 产细菌素乳酸菌的筛选与鉴定[J]. 食品工业科技, 2017, (17): 146-151. DOI: 10.13386/j.issn1002-0306.2017.17.028
LIU Ying-li, DING Li, XIE Liang-xu, WANG Jing, ZHANG Hui-juan. Screening and identification of bacteriocin produced by lactic acid bacteria[J]. Science and Technology of Food Industry, 2017, (17): 146-151. DOI: 10.13386/j.issn1002-0306.2017.17.028
Citation: LIU Ying-li, DING Li, XIE Liang-xu, WANG Jing, ZHANG Hui-juan. Screening and identification of bacteriocin produced by lactic acid bacteria[J]. Science and Technology of Food Industry, 2017, (17): 146-151. DOI: 10.13386/j.issn1002-0306.2017.17.028

产细菌素乳酸菌的筛选与鉴定

Screening and identification of bacteriocin produced by lactic acid bacteria

  • 摘要: 利用溶钙圈法及牛津杯双层平板法,从欧洲萨拉米香肠、新疆奶酪、自发酵泡菜和韩式辣白菜中分离筛选产细菌素的乳酸菌,通过形态学、生理生化鉴定、16S rDNA将菌株鉴定到种,并对其细菌素基因进行克隆,确定细菌素种类。结果表明:从分离纯化到的206株乳酸菌中筛选出抑菌效果较好的菌株SA102、SA104、N105、N107、N108,在排除酸、过氧化氢等干扰因素后,菌株所产抑菌物质对蛋白酶敏感,初步判定起抑菌作用的是细菌素。16S r DNA结果表明这5株产细菌素的菌株2株为乳酸乳杆菌,3株为植物乳杆菌,通过设计引物对菌株细菌素基因进行PCR快速筛选,结果表明菌株N107、N108含有IIb类细菌素Plantaricin KJ基因编码序列,生理生化指标测试证实这些产细菌素菌株不产气、不产氨、不产硫化氢、不产粘、不产色素,具有较好的生物安全性。最后对发酵液中细菌素粗提物的酸碱稳定性和温度稳定性进行了初步探究,以期为将来细菌素的进一步实际应用提供一定的理论借鉴。 

     

    Abstract: In order to obtain bacteriocin producing strains, lactic acid bacterias were isolated from European salami sausages, Xinjiang cheeses, self fermented pickles, Korean cabbages in chili sauce by using calcium dissolving ring and Oxford cup double plate methods. The strains were identified to species on the basis of morphological, physiological, biochemical characteristics and 16 S r DNA sequence analysis, and the bacteriocin related gene was analyzed to determine bacteriocin types.The results showed that 5 strains ( SA102, SA104, N105, N107, N108) had better antibacterial activity among the 206 strains.The antimicrobial role of this bacteriocin was confirmed by eliminating the interference of organic acid, hydrogen peroxide and by conducting protease sensitivity. Two of these 5 strains were Lactobacillus casei and three were Lactobacillus plantarum.The rapid screening of PCR amplification showed that N107, N108 contained IIb plantaricins JK gene.Physiological and biochemical tests confirmed that 5 strains with good biological safety produced no gas, ammonia, hydrogen sulfide, viscosity and pigment.p H stability and temperature stability of the crude extract of fermentation were explored to provide some information and resources for the further study of bacteriocin.

     

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