天然产物研究与开发 ›› 2019, Vol. 31 ›› Issue (10): 1799-1806.doi: 10.16333/j.1001-6880.2019.10.019

• 开发研究 • 上一篇    下一篇

曼氏无针乌贼缠卵腺抗菌糖蛋白提取工艺优化

陈淑敏,张玉琦,龚芳芳,王晔,刘英丹,孟天宇,娄永江*,李勇勇*   

  1. 宁波大学食品与药学学院,宁波 315211
  • 出版日期:2019-10-28 发布日期:2019-11-11
  • 基金资助:

    国家海洋经济示范项目(NBHY-2017-S2)

Optimization of the extraction technology of antibacterial glycoproteins from the nidamental gland of Sepiella maindroni

CHEN Shu-min,ZHANG Yu-qi,GONG Fang-fang,WANG Ye,LIU Ying-dan,MENG Tian-yu,LOU Yong-jiang*,LI Yong-yong*   

  1. College of food and medicine,Ningbo University,Ningbo 315211,China
  • Online:2019-10-28 Published:2019-11-11

摘要: 为优化曼氏无针乌贼缠卵腺抗菌糖蛋白的提取工艺,探索糖蛋白的抗菌活性。本研究在单因素试验基础上,通过响应面试验对糖蛋白的提取工艺进行优化,同时对粗糖蛋白采用阴离子交换柱(DEAE-52)层析进行分离纯化,并进行电泳鉴定。结果表明,糖蛋白的提取优化参数为:NaOH浓度为0.43 mol/L、提取时间2.8 h、提取温度27 ℃,液料比30∶1,在此条件下,得出抗菌粗糖蛋白实际得率为8.31%±0.18%。粗糖蛋白经阴离子交换柱(DEAE-52)层析有效分离获得组分2和组分3等2个糖蛋白组分,经SDS-PAGE电泳表明2个糖蛋白中的活性蛋白成分均为单一蛋白。结果同时表明,组分2与组分3对大肠杆菌与金黄色葡萄球菌具有非常显著的抑菌效果(P<0.01),且抗菌活性远强于同浓度山梨酸钾(P<0.01)。本研究可为曼氏无针乌贼缠卵腺糖蛋白在生物防腐剂方面的开发利用提供理论基础。

关键词: 曼氏无针乌贼, 缠卵腺, 糖蛋白, 抗菌活性

Abstract: To optimize the extraction condition for the antibacterial glycoproteins from the nidamental gland of the Sepiella maindroni and explore the antibacterial activity of the glycoproteins,this paper optimized the extraction process of glycoprotein by response surface test on the basis of single factor test.At the same time,the crude glycoprotein was separated and purified by DEAE-52 Cellulose ion exchange column chromatography and identified by electrophoresis.The results showed that the optimized parameters were 0.43 mol/L of NaOH,extraction times of 2.8 h,extraction temperature of 27 ℃,and liquid to material ratio was 30∶1.In this condition,the actual antibacterial crude glycoprotein yield was 8.31%±0.18%.Two glycoprotein components,component 2 and component 3,were efficiently separated by DEAE-52 Cellulose ion exchange column chromatography.The SDS-PAGE electrophoresis showed that the active protein components of the two glycoproteins were single proteins.The results also showed that component 2 and component 3 had very significant antibacterial effect against Escherichia coli and Staphylococcus aureus(P<0.01),and the antibacterial activity was much stronger than the same concentration of potassium sorbate (P<0.01).This study can provide a theoretical basis for the development and utilization of glycoproteins from nidamental gland of the Sepiella maindroni in biological preservatives.

Key words: Sepiella maindroni, nidamental gland, glycoproteins, antibacterial activity

中图分类号: 

R284.2