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中国精品科技期刊2020
袁兰云, 王亚璐, 宁喜斌, 刘志东. 南极磷虾中产抗菌活性物质菌株的筛选及系统发育分析[J]. 食品工业科技, 2017, (05): 179-184. DOI: 10.13386/j.issn1002-0306.2017.05.025
引用本文: 袁兰云, 王亚璐, 宁喜斌, 刘志东. 南极磷虾中产抗菌活性物质菌株的筛选及系统发育分析[J]. 食品工业科技, 2017, (05): 179-184. DOI: 10.13386/j.issn1002-0306.2017.05.025
YUAN Lan-yun, WANG Ya-lu, NING Xi-bin, LIU Zhi-dong. Screening and phylogenetic analysis of antibacterial strains isolated from Antarctic Krill[J]. Science and Technology of Food Industry, 2017, (05): 179-184. DOI: 10.13386/j.issn1002-0306.2017.05.025
Citation: YUAN Lan-yun, WANG Ya-lu, NING Xi-bin, LIU Zhi-dong. Screening and phylogenetic analysis of antibacterial strains isolated from Antarctic Krill[J]. Science and Technology of Food Industry, 2017, (05): 179-184. DOI: 10.13386/j.issn1002-0306.2017.05.025

南极磷虾中产抗菌活性物质菌株的筛选及系统发育分析

Screening and phylogenetic analysis of antibacterial strains isolated from Antarctic Krill

  • 摘要: 分别以金黄色葡萄球菌Staphylococcus aureus ATCC 27661(Sa 27661)和大肠杆菌Escherichia coli ATCC 35218(Ec 35218)为指示菌,以南极磷虾中分离出的59株菌为测试菌,利用牛津杯法进行抗菌活性菌株的初筛及复筛,同时用比浊测定法和牛津杯法对活性菌株的部分特性包括生长曲线、抗菌曲线、最佳温度、最佳p H、最佳盐度进行了测定,以Mega 5邻接法对其16S r RNA基因序列及系统发育地位进行探究。结果发现,初筛获得13株菌对Sa 27661具有抗菌活性,6株菌对Ec 35218具有抗菌活性。复筛得到对Sa 27661和Ec 35218均具有明显抗菌活性的菌株3株,阳性率为5.08%,其编号分别为1A3、A4、1A5。且3株菌分别培养16、20、28 h进入生长对数期,培养36、48、60 h达到稳定期。分别培养56、64、72 h时抗菌活性达到最高,之后基本达到稳定。最佳温度分别为15、20、20℃,最佳p H分别为7、6、6,最佳盐度分别为1%、3%、2%。系统发育分析结果表明,菌株A4、1A5,均属于嗜冷杆菌属Psychrobacter,菌株1A5与已注册的菌株(Psychrobacter sp.OK1)基因序列相似性可达100%。而菌株1A3与不可培养嗜冷杆菌属(Uncultured Psychrobacter)同源性为97%。三株菌均具有较好的耐酸性、耐碱性和耐盐性的能力,同时具有较好的抗菌活性。 

     

    Abstract: Staphylococcus aureus ATCC 27661 and Escherichia coli 35218 were used as indicator strains respectively.Screening of the antibacterial activity strains from the 59 strains isolated from Antarctic Krill was by the method of Oxford cup diffusion.The partial characteristics including growth curve, antibacterial curve, optimum temperature, optimum p H and optimum salinity of the antibacterial strains were also studied by the method of nephelometry and Oxford cup diffusion.And with the method of Mega 5 adjacency, the 16S rRNA gene sequence of the antibacteria strains were blasted and then the phylogenetic tree were constructed.The results showed that thirteen strains of Sa 27661 antibacteria activity and six strains of Ec 35218 antibacteria activity were screened in the preliminary screening.Also three antibacterial strains named as 1A3, A4, 1A5 both of Sa 27661and Ec 35218 antibacteria activities were screened in the dual-frequency screening.The positive rate was 5.08%.And they reach the logarithmic phase when they were cultured 16, 20, 28 hours and reach the stationary phase when they were cultured 36, 48, 60hours respectively.They have the highest antibacterial activities when they were cultured 56, 64, 72 hours respectively.The optimum temperature of the antibacterial activity strains were 15℃, 20℃, 20℃, optimum pH values were 7, 6, 6 and optimum salinity were 1%, 3%, 2%respectively.Phylogenetic analysis indicated that strains of A4, 1A5 belong to genera of Psychrobacter.And the gene sequence similarity of strain 1A5 was up to 100%of the registered Psychrobacter sp.OK1 (AB302184.1) .But the homology of strain 1A3 was only 97%to Uncultured Psychrobacter which need further study.Three strains all have the ability of acid resistance, alkali resistance, salt tolerance, also the antibacteria activity which need further study.

     

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