天然产物研究与开发 ›› 2011, Vol. 23 ›› Issue (4): 612-619.

• 研究论文 • 上一篇    下一篇

苦瓜籽蛋白的PEG/(NH4)2SO4双水相分离及抑菌作用的研究

蔡 马1,于 群2,朱新产2?   

  1. 1仲恺农业工程学院 生命科学学院,广州 510225;2青岛农业大学 生命科学学院,青岛 266109
  • 收稿日期:2010-10-08 出版日期:2011-08-25 发布日期:2011-09-13
  • 基金资助:

    青岛农大-仲恺农工学院合作研发基金(620325)

Isolation of Momordica charantia Seed Proteins Using Polyethylene Glycol/Ammonium Sulfate Aqueoustwo-phase System and Its Antimicrobial Activity

CAI Ma1, YU Qun2, ZHU Xin-chan2*   

  1. 1College of Life Sciences, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China; 2College of Life Science Qingdao Agricultural University, Qingdao 266109, China
  • Received:2010-10-08 Online:2011-08-25 Published:2011-09-13

摘要: 优化试验条件影响因素,建立PEG/(NH4)2SO4 双水相系统,确定出分离苦瓜籽蛋白的最适PEG6000和(NH4)2SO4 质量分数分别是22%、25%,分配系数为0.089,回收率为96%。双水相分离的苦瓜籽蛋白经SDS-PAGE凝胶电泳显示,主要蛋白质是50 KD、43 KD和10 KD亚基结合形成的多聚体12S(327 KD)球蛋白,还原条件下出现诸多小于35 kD的蛋白组分,表明蛋白质亚基是通过二硫键结合。抑菌试验显示苦瓜籽蛋白对受试细菌菌和真菌产生明显的抑菌活性,其MIC远低于苯甲酸钠,而且在不同盐离子浓度、温度、pH及紫外线等因素处理时,均具有一定的稳定性。此外试验还表明苦瓜籽蛋白也具有较强的清除羟自由基能力和总抗氧化能力。

Abstract: The Momordica charantia seed proteins were extracted and separation by polyethylene glycol/ammonium sulfate aqueoustwo-phase system. The factors affecting system distribution coefficients, recycling rate, and extracting agents were investigated and the optimal conditions of M. charantia seed proteins extraction were confirmed. Under the conditions of mass fraction of PEG6000 as 22% and (NH4)2SO4 as 25%, the smallest distribution coefficient obtained as 0.089 and the recovery was 96%. The results of SDS-PAGE showed that M. charantia seed protein were polymer globulin of 12S (327 KD) with major subunits 50 KD, 43 KD, and 10 KD, which were connected with disulfide bonds by low molecular weight protein subunits (<35kD). The results of antimicrobial activities analysis showed that the proteins had strong inhibition to testing bacteria and fungi, whose MIC were far below the value of sodium benzoate, and which were very stable under the environment of low temperature, acid, ultraviolet, and inorganic salt. In addition, the test also showed total antioxidative activity and clearance on •OH free radical of the proteins were obvious. Therefore, the M. charantia seed proteins had the significant antioxidant capacity and broad-spectrum antimicrobial activity.

Key words: Momordica charantia seed protein, aqueous two-phase system, extraction and separation, antimicrobial activities, antioxidant activity.