天然产物研究与开发 ›› 2014, Vol. 26 ›› Issue (12): 1921-1925.

• 研究论文 • 上一篇    下一篇

条斑紫菜蛋白酶解产物中抗菌肽的纯化及特征

刘冬冬,周锋,周楠迪,田亚平*   

  1. 江南大学工业生物技术教育部重点实验室,无锡 214122
  • 出版日期:2014-12-31 发布日期:2015-01-09

Purification and Characterization of Antimicrobial Peptides from the Proteolytic Product of Laver Protein

LIU Dong-dong, ZHOU Feng, ZHOU Nan-di, TIAN Ya-ping*   

  1. Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China
  • Online:2014-12-31 Published:2015-01-09

摘要: 以酶解产物对金黄色葡萄球菌的抑菌能力为指标,筛选出木瓜蛋白酶并优化确定酶解条件,酶解液中分子量在1000 Da以下的小分子肽类占96.68%;采用DEAE-52纤维素阴离子交换层析、Sephadex G-25凝胶层析得到纯化的紫菜蛋白抗菌肽(LPAP);液质联用分析该纯化抗菌肽中主要的多肽序列为FFDD(Phe-Phe-Asp-Asp);该LPAP对G+菌和G-菌都有抑制作用,具有相对广谱性,对金黄色葡萄球菌的最小抑菌浓度(Minimum inhibitory concentration,MIC)为0.25 mg/mL,抗菌肽作用金黄色葡萄球菌的电镜照片表明其抑菌机制可能是对细胞膜的破坏作用。

关键词: 紫菜, 蛋白酶, 水解产物, 抑菌肽, 分离纯化

Abstract: In this study,papain was chosen as the hydrolyzing enzyme and the enzymatic hydrolysis conditions were optimized with the inhibitory effect of protein hydrolysate against Staphylococcus aureus as index.The relatively molecular weight distribution of laver enzymolysis liquid showed that laver protein with molecular weight below 1000 Da accounted for 96.68%.The laver protein antimicrobial peptides(LPAP)was purified through DEAE-52 cellulose anion exchange chromatography and Sephadex G-25 gel filtration chromatography,which was identified as a tetrapeptide FFDD(Phe-Phe-Asp-Asp)by Q-TOF-MS.This LPAP possessed a relatively broad spectrum of antibacterial activity,and showed significant growth inhibition against gram-positive bacterials and gram-negative bacterials.Minimum inhibitory concentration of LPAP against S.aureus was 0.25 mg/mL.The electron microscope photos of S.aureus impacted by antibacterial peptides revealed its antibacterial mechanism was mainly due to the destruction of cell membrane.

Key words: laver, protease, hydrolyzate, antimicrobial peptides, purification

中图分类号: 

Q946.1 TS254.9