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  

Study on the anti-microbial and anti-tumor activities of the secondary metabolites produced by a symbiotic or epiphyte Bacillus subtilis 110 fromSinonovacula Constricta(Lamarck)

作　　者：张付云[1] 王玟苏 钟键[1] 孙秋颖[1] 赵宇[1] 祁艳霞[1] 

机构地区：[1]大连海洋大学食品科学与工程学院,辽宁大连116023

出　　处：《中国海洋药物》2014年第6期15-20,共6页Chinese Journal of Marine Drugs

基　　金：大连市科学技术基金项目(2010J21DW017);辽宁省大学生创新创业训练项目(201310158204);大连海洋大学2012年校列科研项目(2012HYDX15)资助

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0.22μm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25kDa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20mm。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200μg/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。Objective To detect the anti-microbial active component of the secondary metabolites produced by symbiotic or epiphyte bacterium 110 from Sinonovacula Constricta （Lamarck）, and preliminarily study the anti-tumor effects of the anti-microbial active component. Methods The thalli of symbiotic or epiphyte bacterium 110 were separated from its fermentation broth by the 0. 22 /~m 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, were 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. Results The antibacterial activity of fermentation broth from 110 was much higher than the antibacterial activity of thalli. The Sediment of fermentation broth precipitated by am- monium sulfate showed a strong antibacterial activity, and the Sediment was mainly composed of 60, 45 and 25 kDa three kinds of protein. The etude protein 110 demonstrated inhibitory activity to 8 kinds of fungi, and the diameter of inhibition zone to Ralstonia solanacearurn, Botrytis cinerea and Fusariurn o.rysporurn Schlecht was more than 20 ram. The crude protein also exhibited inhibition activity to leu- kemia cell K562, hepatoma cell HepG-2 and breast cancer cell MDA-MB-231. The inhibition rate to hu- man hepatoma cell HepG-2 reached up to 74. 01% when the concentration of this etude protein was 200 /~g / mL. Conclusion The extracellular protein of symbiotic or epiphyte bacterium 110 from Sinonovacu- Za Constricta （Lamarck） has potential development value for its anti-tumor activity and broad-spectrum antimicrobial activity.

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 

分 类 号：R931[医药卫生—生药学]