食品科学 ›› 2019, Vol. 40 ›› Issue (24): 258-264.doi: 10.7506/spkx1002-6630-20181112-123

• 工艺技术 • 上一篇    下一篇

辣椒籽抗菌肽提取条件优化及分离纯化

韩玉竹,李平兰,何琴,赵展展,刘雪文,杨叶梅   

  1. (1.西南大学动物科学学院,重庆 402460;2.中国农业大学食品科学与营养工程学院,北京食品营养与人类健康高精尖创新中心,北京 100083;3.重庆市畜牧科学院,重庆 402460)
  • 出版日期:2019-12-25 发布日期:2019-12-24
  • 基金资助:
    “十二五”国家科技支撑计划项目(2015BAD16B01-4);中央高校基本科研业务费专项(XDJK2018B024); 重庆市技术创新与应用示范项目(cstc2018jscx-msyb0430)

Optimization of Extraction Conditions, Separation and Purification of Antifungal Peptides from Hot Pepper Seeds

HAN Yuzhu, LI Pinglan, HE Qin, ZHAO Zhanzhan, LIU Xuewen, YANG Yemei   

  1. (1. College of Animal Science, Southwest University, Chongqing 402460, China; 2. Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China; 3. Chongqing Academy of Animal Sciences, Chongqing 402460, China)
  • Online:2019-12-25 Published:2019-12-24

摘要: 以黄曲霉为指示菌,抑菌圈直径为考察指标,比较不同辣椒籽蛋白提取物的抑菌效果。结果显示,6 种辣椒籽均对黄曲霉生长有抑制作用,其中贵州灯笼椒的抑菌效果最好。以该品种辣椒籽为后续实验原料,对其提取条件进行优化。获得最佳提取条件为:浸提液为乙二胺四乙酸缓冲液,料液比1∶5.05(g/mL)、浸提时间16.13 h、硫酸铵饱和度81.15%。在此优化条件下,辣椒籽抗菌肽抑菌圈直径可达23.89 mm,比优化前(12.34 mm)提高了93.06%。结合透析袋脱盐、10 kDa超滤管超滤以及葡聚糖凝胶层析,对辣椒籽抗菌肽进行分离纯化,纯化后的抗菌肽经Tricine-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测已达到电泳纯,分子质量在7.8 kDa左右。

关键词: 辣椒籽, 抗菌肽, 提取条件, 分离纯化

Abstract: Protein extracts from the seeds of different cultivars of hot pepper were screened for their antifungal activity against Aspergillus flavus in terms of inhibition zone diameter. The results showed that the seeds of all six cultivars investigated had inhibitory effects on the growth of A. flavus, and Guizhou lantern pepper seeds had the strongest antifungal effect of them. The conditions for extraction of antifungal proteins from the Guizhou Lantern cultivar were optimized as follows: EDTA buffer as the extraction solvent, solid-to-solvent ratio 1:5.05 (g/mL), extraction time 16.13 h, and ammonium sulfate saturation 81.15%, leading to an inhibitory zone diameter of 23.89 mm, which was 93.06% higher than that (12.34 mm) before optimization. In addition, the antifungal peptides from hot pepper seeds were separated and purified by dialysis, 10 kDa ultrafiltration and Sephadex gel chromatography. The purified antifungal peptide was found to be homogenous with a molecular mass of around 7.8 kDa by Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

Key words: hot pepper seeds, antimicrobial peptides, extraction conditions, separation and purification

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