张付云,王玟苏,钟键,等.螠蛏共附枯草芽孢杆菌110次级代谢产物的抑菌抗肿瘤活性研究Δ[J].中国海洋药物,2014,33(6):15-20. |
螠蛏共附枯草芽孢杆菌110次级代谢产物的抑菌抗肿瘤活性研究Δ |
Study on the anti-microbial and anti-tumor activity of the secondary metabolites produced by a symbiotic or epiphyte Bacillus subtilis 110 from Sinonovacula Constricta (Lamarck) |
投稿时间:2014-07-30 修订日期:2014-09-04 |
DOI: |
中文关键词: 螠蛏 共附生细菌 次级代谢产物 抑菌活性 抗肿瘤活性 |
English Keywords:Sinonovacula Constricta (Lamarck) Symbiotic or epiphyte bacterium Secondary metabolites Anti-microbial activity anti-tumor activity |
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中文摘要: |
摘要:目的 检测螠蛏共附枯草芽孢杆菌110次级代谢产物的抑菌活性组分,并探讨抑菌活性组分的抗肿瘤作用。方法 用0.22 μm滤膜过滤将螠蛏共附枯草芽孢杆菌110的菌体和发酵液分开,用硫酸铵沉淀发酵液的蛋白组分并用SDS-PAGE检测,用滤纸片法分别检测菌株发酵液、菌体及胞外蛋白的抑菌活性,琼脂块法检测抑菌活性组分的抑真菌活性,用海虾毒性法和MTT法检测抑菌活性组分的抗肿瘤活性。结果 螠蛏共附枯草芽孢杆菌110的发酵液抑菌活性远远高于菌体的抑菌活性,发酵液的硫酸铵沉淀物具有很强的抑菌活性,硫酸铵沉淀的110活性粗蛋白主要由分子量约为60、45和25 kDa的3条带组成。110活性粗蛋白对8种真菌指示菌具有抑制作用,其中对辣椒青枯、番茄灰霉和瓜类枯萎的抑菌圈直径超过20 mm。该活性粗蛋白还对白血病细胞K562、肝癌细胞HepG-2和人乳腺癌细胞MDA-MB-231具有抑制作用,其中200 μg/mL的蛋白对人肝癌细胞HepG-2抑制率达74.01%。结论 螠蛏共附枯草芽孢杆菌110胞外蛋白有抗肿瘤活性和广谱抑菌活性,具有潜在的开发价值。 |
English Summary: |
Abstract: Objective To detect the antimicrobial active component of the secondary metabolites produced by symbiotic or epiphyte bacterium 110 from Sinonovacula Constricta (Lamarck), and Preliminary study the antitumor effects of the antimicrobial active component. Methods The thalli of symbiotic or epiphyte bacterium 110 was separated from its fermentation broth by the 0.22 μm filter membrane. The protein in the fermentation broth was precipitated by ammonium sulfate and analyzed by SDS-PAGE. The antibacterial activity of fermentation broth, thalli and extracellular proteins, was detected by disc diffusion method. The antifungal activity of antimicrobial active component was assayed by Agar block technique. The anti-tumor activity of antimicrobial active component was tested by shrimp toxicity and MTT assay. Result The antibacterial activity of fermentation broth from 110 is much higher than the antibacterial activity of thalli. The Sediment of fermentation broth precipitated by ammonium sulfate showed a strong antibacterial activity, and the Sediment was mainly composed of 60, 45 and 25 kDa three kinds protein. The crude protein 110 demonstrated inhibit activity to 8 kinds of fungi, and the diameter of inhibition zone to Ralstonia solanacearum, Botrytis cinerea and Fusarium oxysporum Schlecht is more than 20 mm. The crude protein also exhibit inhibition activity to leukemia cell K562, hepatoma cell HepG-2 and breast cancer cell MDA-MB-231. The inhibition rate to human hepatoma cell HepG-2 reached up to 74.01% when the Concentration of this crude protein was 200 μg / mL. Conclusion The extracellular protein of symbiotic or epiphyte bacterium 110 from Sinonovacula Constricta (Lamarck) has potential development value for its anti-tumor activity and broad-spectrum antimicrobial activity. |
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