四川农业大学学报 ›› 2007, Vol. 25 ›› Issue (03): 271-276.doi: 10.16036/j.issn.1000-2650.2007.03.005

• 研究论文 • 上一篇    下一篇

仙人掌抗菌肽的筛选、分离纯化及性质研究

刘一江, 黄乾明, 杨洲平, 陈平, 杨菲   

  1. 四川农业大学 生命科学与理学院, 四川 雅安 625014
  • 收稿日期:2007-03-26 出版日期:2007-09-30 发布日期:2017-03-04
  • 基金资助:
    四川省科学技术厅应用基础基金(04JY029-032);四川省教育厅重点科研基金(2002A009,2004A026);四川农业大学科技创新基金(00731200)。

Screening of Antibacterial Peptides from Cactuses and Characterization of Antimicrobial Peptide MH-AMP-1 Purified from Mammillaria Haw

LIU Yi-jiang, HUANG Qian-ming, YANG Zhou-ping, CHEN Ping, YANG Fei   

  1. College of Biology and Science, Sichuan Agricultural University, Yaan 625014, Sichuan, China
  • Received:2007-03-26 Online:2007-09-30 Published:2017-03-04

摘要: 目的:研究仙人掌植物是否具有起抑菌作用的抗菌肽,并对其进行分离纯化和部分性质研究。方法:采用梯度盐析的方法提取6种仙人掌不同饱和梯度硫酸铵沉淀的蛋白质组分进行抑菌性试验,筛选蛋白质组分具有抗菌效果的仙人掌品种。用Sephadex G-75和Sephadex G-25柱层析对抗菌效果最为显著的蛋白质组分进一步分离纯化,并对纯化到的具有抗菌活性的多肽物质进行分子量、理化性质和抗菌活性检测。结果:发现4种仙人掌中存在对金黄色葡萄球菌有抗菌作用的小分子量蛋白质成分。其中金手球所含蛋白质组分的抗菌效果最为显著,并从中分离纯化得到具有抗菌活性的多肽物质,命名为MH-AMP-1。MH-AMP-1的分子量(Mr)约为8511 Da,在pH 2.5~7.5的乙酸铵缓冲液中抗菌活性强,且具有一定的热稳定性,对检测的病原菌具有广谱抗菌活性。

关键词: 仙人掌, 抗菌肽, 分离纯化

Abstract: To search the existence of antimicrobial peptides in cactus, the peptide was isolated and purified for characterization study. The proteins of six kinds of cactus were isolated by sulfate precipitation on different saturation ranges and their antimicrobial activities were tested to screen the cactus with antimicrobial peptides. The Sephadex G-75 and Sephadex G-25 were used to further purify the peptide with most effective antimicrobial activity. SDS-PAGE, Proline and Ninhydrin Reaction, Resistance-bacterial test, et al were used to analyze its characterization. The results show ed that four kinds of Cactus had low molecular weight proteins with antimicrobial activities and the proteins extracted from Mammillaria Haw. were most effective on antimicrobial activity. An antimicrobial peptide, named MH-AMP-1, was isolated from the Mammillaria Haw. The molecular weight of MH-AMP-1 was about 8511 Da and its antimicrobial activity remained potent in the buffer with pH 2.5~7.5. MH-AMP-1 was thermostable to some extent and could kill most pathogens.

Key words: Cactus, antimicrobial peptide, isolation and purification

中图分类号: 

  • Q949.759.9