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中国东海亚硫酸杆菌属微生物的鉴定及生物学活性分析
龙聪,卢小玲,刘军华,高云,焦炳华*,刘小宇*
0
(第二军医大学基础部生物化学与分子生物学教研室,上海 200433)
摘要:
目的:对44株中国东海来源的亚硫酸杆菌进行16S rDNA的序列测定和生物学活性的初步分析。方法:用PCR方法对细菌的16S rDNA序列进行扩增,并在GenBank上进行相似性搜索。将全部亚硫酸杆菌的16S rDNA序列运用Clustal X1.8软件进行多序列比对并用MEGA4.1绘制进化树。利用稻瘟霉菌(Pyricularia oryzae) P-2b、枯草芽孢杆菌 (Bacillus subtilis) 和大肠埃希菌 (Escherichia coli) 作为指示菌检测菌株发酵液的抗菌活性。以HL-60、HepG2细胞作为细胞毒活性指示细胞株检测菌株发酵液的细胞毒活性。采用ABTS和DPPH抗氧化模型检测菌株发酵液的自由基清除能力。结果:经与GenBank数据库比对,44株菌均与亚硫酸杆菌属有很高的相似性(97%~100%)。所有菌株发酵液对指示菌未显示出抗菌活性。菌株发酵液有不同程度的细胞毒活性和清除自由基的能力。19株菌的发酵液对HepG2细胞有抑制作用,抑制率为6.8%~42.8%,18株菌的发酵液对HL-60细胞有抑制活性,抑制率为6.9%~43.6%。12株菌的发酵液对ABTS自由基有清除能力,清除率为4.49%~23.08%,8株菌发酵液对DPPH自由基有清除能力,清除率为1.23%~30.76%。结论:44株中国东海来源的亚硫酸杆菌表现出显著的细胞毒活性及抗氧化活性,具有潜在的药用价值。
关键词:  亚硫酸杆菌  鉴定  细胞毒性  自由基清除
DOI:10.3724/SP.J.1008.2009.01106
投稿时间:2009-07-20修订日期:2009-09-21
基金项目:国家高技术研究发展计划 (“863” 计划,2006AA09Z416,2006AA09Z425,2007AA091501).
Identification and biological activity screening of Sulfitobacter sp.from East China Sea
LONG Cong,LU Xiao-ling,LIU Jun-hua,GAO Yun,JIAO Bing-hua*,LIU Xiao-yu*
(Department of Biochemistry and Molecular Biology,College of Basic Medical Sciences,Second Military Medical University,Shanghai 200433,China)
Abstract:
Objective:To determine the 16S rDNA sequences of 44 strains of Sulfitobacter sp. obtained from the East China Sea and to study their biological activities. Methods:PCR was used to amplify the 16S rDNA sequences of the 44 strains,and the obtained sequences were subjected to GenBank similarity search.The multiple sequence alignment of the 44 strains was done by Clustal X1.8 and MEGA4.1.The antibacterial activities of Sulfitobacter sp. fermentation broths were tested using the following indicator bacteria:Pyricularia oryzae,Bacillus subtills,and Escherichia coli.The cytotoxic activities of the fermentation broths were tested using HL-60 and HepG2 as the indicator cells.ABTS and DPPH antioxidant models were applied to test the free radical scavenging efficacy of the fermentation broth.Results:Analysis of the 16S rDNA genes of the 44 strains showed high similarity (97%-100%) to Sulfitobacter sp..No fermentation broths of the 44 strains showed evident anti-bacterial activities,and they showed different degrees of cytotoxic activities and free radical scavenging capacities.Nineteen strains inhibited the growth of HepG2 cells,with the inhibition rate ranging 6.8%-42.8%; 18 strains inhibited the growth of HL-60 cells,with the inhibition rate ranging 6.9%-43.6%; 12 strains showed ABTS radical scavenging capacity,with the elimination rate ranging 4.49%-23.08%; and 8 strains showed DPPH radical scavenging capacity,with the elimination rate ranging 1.23%-30.76%.Conclusion:The 44 Sulfitobacter sp.strains possess evident cytotoxic activities and free radical scavenging capacities,making them potential for medicinal application and laying a foundation for further development of these bacteria.
Key words:  Sulfitobacter sp.  identification  cytotoxicity  free radical elimination