论文部分内容阅读
目的:对44株中国东海来源的亚硫酸杆菌进行16SrDNA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16SrDNA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16SrDNA序列运用ClustalX1.8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。
OBJECTIVE: To determine the sequence of 16S rDNA and the biological activity of sulphites in 44 strains of East China Sea. Methods: The bacterial 16S rDNA sequence was amplified by PCR and searched for similarity in GenBank. Multiplex alignments of all 16SrDNA sequences of sulfinobacillus were performed using the ClustalX 1.8 software and the phylogenetic tree was constructed using MEGA 4.1. Pyricularia oryzae P-2b, Bacillus subtilis and Escherichia coli were used as indicator bacteria to test the antibacterial activity of the fermentation broth. Using HL-60, HepG2 cells as cytotoxic activity indicator cell line detection of strain fermentation broth cytotoxic activity. ABTS and DPPH antioxidant model was used to detect the free radical scavenging capacity of the strain fermentation broth. Results: Compared with the GenBank database, 44 strains were highly similar to the sulfatobacilli (97% -100%). All strains of broth showed no antibacterial activity against the indicator bacteria. Strain fermentation broth with varying degrees of cytotoxic activity and the ability to scavenge free radicals. The fermentation broth of 19 strains could inhibit HepG2 cells with the inhibitory rate of 6.8% ~ 42.8%. The fermentation broth of 18 strains had inhibitory activity of 6.9% ~ 43.6% on HL-60 cells. The fermentation broth of 12 strains could scavenge ABTS free radicals with the clearance rate of 4.49% -23.08%. The eight strains of fermentation broth scavenging DPPH free radicals with the clearance rate of 1.23% -30.76%. CONCLUSIONS: 44 strains of sulfatobacillus sulphites from the East China Sea show remarkable cytotoxicity and anti-oxidative activity with potential medicinal value.