食品科学

• 基础研究 • 上一篇    下一篇

银杏种仁蛋白分离纯化及其抑菌活性

吴海霞,吴彩娥,范龚健,李婷婷,应瑞峰,华 菁   

  1. 1.南京林业大学森林资源与环境学院,江苏 南京 210037;2.运城学院生命科学系,山西 运城 044000;
    3.南京林业大学轻工科学与工程学院,江苏 南京 210037
  • 出版日期:2014-07-15 发布日期:2014-07-18

Purification and Antimicrobial Activity of Protein from Ginkgo biloba Seed Kernels

WU Hai-xia, WU Cai-e, FAN Gong-jian, LI Ting-ting, YING Rui-feng, HUA Jing   

  1. 1. College of Forest Resources and Environment, Nanjing Forestry University, Nanjing 210037, China;
    2. Department of Life Science, Yuncheng University, Yuncheng 044000, China;
    3. College of Light Industry Science and Engineering, Nanjing Forestry University, Nanjing 210037, China
  • Online:2014-07-15 Published:2014-07-18

摘要:

从银杏种仁中分离纯化具有抑菌活性的蛋白质,并分析其抑菌活性。结合抑菌活性分析,采用硫酸铵分级盐析、透析、纤维素DE-52阴离子交换柱层析及Sephadex G-75凝胶过滤柱层析对银杏种仁蛋白(Ginkgo bilobaseed protein,GBSP)进行分离纯化,得到一种抑菌蛋白GBSPⅠ-A。十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(sodiumdodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)检测结果显示,该蛋白呈单一条带,其分子质量约为42.80 kD;Superdex G-75柱层析结果显示GBSPⅠ-A呈单一对称峰,高效凝胶渗透色谱测定其分子质量为39.32 kD;该蛋白对肺炎克雷伯氏菌、金黄色葡萄球菌、戴尔有孢圆酵母及黑曲霉的最小抑菌浓度(minimum inhibitory concentration,MIC)分别为20、20、20、12 mg/mL。

关键词: 银杏种仁蛋白, 分离纯化, 抑菌, 分子质量

Abstract:

An antimicrobial protein was isolated and purified from the seed kernels of Ginkgo biloba and its antimicrobial
activity was analyzed. Ginkgo biloba seed protein (GBSP) was purified by ammonium sulfate precipitation, dialysis, DEAEcellulose
DE52 ion-exchange and Sephadex G-75 gel filtration through bioassay-guided method. A protein fraction named
GBSPⅠ-A possessing antimicrobial activity, was obtained. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
(SDS-PAGE) analysis suggested that the protein was homogeneous and its apparent molecular mass was 42.80 kD. The
results of chromatographic identification by Superdex G-75 and HPGPC showed a single and symmetrical peak with a
molecular weight of 39.32 kD. The protein exhibited potent antimicrobial activity against K. pneumoniae, S. aureus,
T. delbrueckii, and A. niger with minimum inhibitory concentrations (MIC) of 20, 20, 20, and 12 mg/mL, respectively.
This study could provide a theoretical basis for the discovery of a new antibacterial protein and the development of
food preservatives.

Key words: Ginkgo biloba seed protein (GBSP), isolation and purification, antimicrobial activity, molecular weight