土壤中芽孢杆菌提取蛋白对致病菌的生物活性研究

H. A. Ibrahim, N. H. Zaki
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摘要

从巴格达市的不同地区收集了40个土壤样本。52株分离物中,芽孢杆菌属38株(73.1%),经培养、镜检、生化及VITEK 2系统检测,蜡样芽孢杆菌最多21株(55.3%),其次为枯草芽孢杆菌11株(28.9%),解淀粉芽孢杆菌6株(15.8%)。蜡样芽孢杆菌(B1)和枯草芽孢杆菌(B16)菌株的抑菌活性最高。提取细胞外蛋白,硫酸铵(80%)饱和纯化,经Sepharose-6B凝胶过滤柱。用sds -聚丙烯酰胺凝胶电泳法测定蛋白质分子量,蜡样芽孢杆菌和枯草芽孢杆菌各有一条条带,分子量分别为68和50 KDa。结果表明,对革兰氏阳性菌有较高的抑菌活性,对革兰氏阴性菌除铜绿假单胞菌外,抑菌活性一般,无效果。该提取物对脓毒杆菌的抗菌效果较好,对金黄色葡萄球菌、大肠杆菌、粘质葡萄球菌、K. spp均有中等效果,对铜绿假单胞菌无效果。细胞毒性实验结果显示,纯化提取物在1000µg/ml浓度下对乳腺癌MCF7细胞具有较高的细胞毒作用,而粗提取物在100µg/ml浓度下对乳腺癌MCF7细胞具有较高的细胞毒作用。
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The Biological Activity of Some Extracted Proteins from Bacillus spp. Isolated from Soil Against Pathogenic Bacteria
Forty soil samples were collected from different regions in Baghdad city. Among 52 isolate, only 38 isolates (73.1 %) belong to Bacillus spp. All bacterial isolates have been submitted to the cultural, microscopical, biochemical examination and VITEK 2 System, Bacillus cereus was predominantly found 21 (55.3%) followed by Bacillus subtilis 11 (28.9%), then Bacillus amyloliquefaciens was 6 (15.8%). Bacillus cereus (B1) and Bacillus subtilis (B16) isolates were with the highest level with the antibacterial activity when selected. Extracellular proteins were extracted and purified with ammonium sulfate (80%) saturation, then submitted to Sepharose-6B gel filtration column. SDS-polyacrylamide gel electrophoresis was used to determine the molecular weights of proteins, and one band appeared to each of Bacillus cereus and Bacillus subtilis with molecular weight 68 and 50 KDa respectively. The results revealed that higher effect of antibacterial activity on Gram positive bacteria, and moderate on Gram negative bacteria except P.aeruginosa with no effect. The antibiofilm effect of the extract found higher effective against S.pyogens, moderate effect against each of (S.aureus, E.coli, S. marcescens, K. spp) and no effect against P.aeruginosa. The results of cytotoxicity showed that the purified extract was a higher cytotoxic effect on breast cancer MCF7 cells at 1000µg/ml concentration, while the crude extract had a higher cytotoxic effect on breast cancer MCF7 cells at 100µg/ml concentration.
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