摘要
为促进江蓠内生真菌NSS1抗菌蛋白的应用研究,利用硫酸铵沉淀法制备抗菌蛋白,最佳硫酸铵饱和度为65%。采用滤纸片法检测其对细菌指示菌的抑菌活性,抗菌粗蛋白均能抑制五种细菌指示菌的生长,当浓度达到750μg/mL时,抑制效果最好。采用邻苯三酚自氧化法测定蛋白对超氧阴离子自由基(O-2)的清除作用,MTT法检测蛋白对肿瘤细胞的影响。抗菌蛋白在100℃以下,pH中性时抑菌活性稳定,对紫外线照射不敏感,丙三醇、甲醇和胰蛋白酶对抗菌蛋白的抑菌活性没有影响。不同浓度的蛋白液对超氧阴离子均有清除效果。当蛋白浓度达到360μg/mL时对肿瘤细胞的抑瘤作用最强。上述结果显示抗菌蛋白具有较强的抑菌活性、抗氧化活性和抗肿瘤活性。
Inorder to study antimicrobail activity of protein from Gracilaria endophytic fungi NSS1, the antimicrobial protein was prepared by ammonium sulfate precipitation, the best ammonium sulfate saturation was 65%. Antibacterial activity to baeterial indicator was detected by filter paper method, the results showed that the antibacterial crude protein could inhibit the growth of five indieator bacteria, and the best inhibitory effect was obtained when the protein concentration reached 750 μg/mL. The scavenging effect of superoxide anion free radical （ O2^- ） from protein was deteeted by the pyrogallol autoxidation method. The effect of protein on tumor cells was detected by the MTT method. The antibacterial activity was stable under the condition of below 100℃ and neutral pH. The activity of protein was not sensitive to UV irradiation, glycerol, methanol and trypsin. Different concentrations of protein solution all had effect on removing superoxide anion. It had the strongest inhibitory effect on tumor ceils, when the protein concentration was 360 μg,/mL. Above results indicated that the antibacterial protein had strong antibacterial activity, antioxidant activity and antitumor activity.
出处
《生物技术进展》
2014年第2期124-128,共5页
Current Biotechnology
基金
辽宁省大连市科学技术基金项目(2010J21DW017)
大连海洋大学2012年校列科研项目(2012HYDX15)资助
关键词
内生真菌
抗菌蛋白
抑菌活性
抗氧化活性
抗肿瘤活性
endophytic fungi
antimicrobial protein
antimicrobial activity
antioxidant activity
antitumor activity
