生物技术进展 ›› 2014, Vol. 4 ›› Issue (2): 124-128.DOI: 10.3969/j.issn.2095-2341.2014.02.09

• 研究论文 • 上一篇    下一篇

江蓠内生真菌NSS1蛋白活性的初步研究

王维才,张付云*,刘晔,卢航,李振,赵宇   

  1. 大连海洋大学食品科学与工程学院, 辽宁 大连 116023
  • 收稿日期:2014-02-19 出版日期:2014-03-25 发布日期:2014-03-13
  • 通讯作者: 张付云,副教授,主要从事海洋微生物资源利用研究。E-mail:zhangfuyun2002@aliyun.com.cn
  • 作者简介:王维才,硕士研究生,研究方向为海洋微生物资源综合利用。E-mail: weicaiwang012@163.com。
  • 基金资助:

    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。

Antimicrobial Activity of Protein from Gracilaria Endophytic Fungi NSS1

WANG Wei-cai, ZHANG Fu-yun, LIU Ye, LU Hang, LI Zhen, ZHAO Yu   

  1. College of Food Science and Engineering, Dalian Ocean University, Liaoning  Dalian 116023, China
  • Received:2014-02-19 Online:2014-03-25 Published:2014-03-13

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750 μg/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360 μg/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。

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

Abstract: 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 bacterial indicator was detected by filter paper method, the results showed that the antibacterial crude protein could inhibit the growth of five indicator bacteria, and the best inhibitory effect was obtained when the protein concentration reached 750 μg/mL. The scavenging effect of superoxide anion free radical (O-2) from protein was detected 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 cells, when the protein concentration was 360 μg/mL. Above results indicated that the antibacterial protein had strong antibacterial activity, antioxidant activity and antitumor activity.

Key words: endophytic fungi, antimicrobial protein, antimicrobial activity, antioxidant activity, antitumor activity