食品科学

• 工艺技术 • 上一篇    下一篇

响应面法优化姜黄素提取工艺及其对类淀粉样蛋白聚集形成的抑制

张 良,杨 松,何剑为,邹志远,贾丽娇,蒋 坤,孟宪军,张慧丽   

  1. 1.辽宁大学生命科学院,辽宁 沈阳 110036;2.沈阳农业大学生物技术学院,辽宁 沈阳 110866;
    3.沈阳军区联勤部药品仪器检验所,辽宁 沈阳 110026;4.沈阳农业大学食品学院,辽宁 沈阳 110866
  • 出版日期:2013-10-25 发布日期:2013-09-28

Optimization of Curcumin Extraction by Response Surface Methodology and Its Inhibitory Effect on Amyloid Aggregation

ZHANG Liang,YANG Song,HE Jian-wei,ZOU Zhi-yuan,JIA Li-jiao,JIANG Kun,MENG Xian-jun,ZHANG Hui-li   

  1. 1. College of Life Science, Liaoning University, Shenyang 110036, China;
    2. College of Biotechnology, Shenyang Agricultural University, Shenyang 110866, China;
    3. Shenyang Joint Logistics Department of Military Institute for Drug and Instrument Control, Shenyang 110026, China;
    4. College of Food Science, Shenyang Agricultural University, Shenyang 110866, China
  • Online:2013-10-25 Published:2013-09-28

摘要:

目的:半胱氨酸酶抑制剂(cC)L68Q突变后能够以二聚体开始进一步产生类淀粉样聚集。研究姜黄素对基因重组突变cC的毕赤酵母Pichia pastoris X-33表达菌的生长和分泌的影响,以及抑制突变cC的类淀粉样聚集的作用。方法:采用响应面法优化姜黄素的提取工艺,通过液体比浊法探究姜黄素对基因重组菌株生长抑制的作用,通过SDS-PAGE研究不同浓度的姜黄素对cC的分泌的作用,通过刚果红染色法研究姜黄素对cC蛋白聚集的抑制作用。结果:由Design-Expert响应面分析软件得到姜黄素最佳提取条件为:液固比30:1(mL/g)、超声功率300W、提取时间15min,此条件下提取率达3.97mg/g。姜黄素对重组菌体生长的抑制作用的相对稳定,但随着姜黄素添加量的增加,其对蛋白分泌量和聚集沉淀量均抑制明显。结论:姜黄素对基因重组cC的生长、分泌和聚集有抑制作用。

关键词: 姜黄素, 半胱氨酸酶抑制剂, 响应面法优化

Abstract:

Objective: The L68Q mutation in the cystatin C (cC) gene can form amyloid through dimers at the initial stage. The
curcumin was used to explore the growth of yeast and the quantity of secretory cC in mutated recombinant yeast Pichia pastoris
as well as its inhibitory effect on amyloid accumulation of cC. Methods: The extraction of curcumin was optimized using response
surface methodology. The inhibitory effect of curcumin on the growth of recombinant cC yeast was measured. The quantity of cC,
secreted by yeast in media with curcumin at different concentrations, was evaluated by SDS-PAGE. The degree of cC amyloid in
the presence of curcumin was explored by Congo red staining. Results: The optimal extraction conditions for solid/solvent ratio,
ultrasonic power and time were 30:1 (mL/g), 300 W and 15 min, respectively. Under these conditions, the yield of curcumin was 3.97
mg/g. The inhibitor effect of curcumin on recombinant yeast growth was relatively mild, but increased curcumin concentration
could more effectively inhibit the secretion of cC and cC amyloid. Conclusion: Curcumin can inhibit the growth, protein secretion
of recombinant yeast and amyloidogenic characteristics of recombinant cystatin.

Key words: curcumin, recombinant cystatin, response surface methodology