摘要
为开发一种简单、有效、低成本的α-淀粉酶抑制剂(α-AI)提取纯化工艺及探索其活性测试最佳条件,本研究以白芸豆为原料,采用水提法从中提取α-AI。首先对3,5-二硝基水杨酸(DNS)微孔法测定α-AI活性的条件进行了研究;随后在单因素基础上,通过正交试验确定了α-AI提取的最佳工艺条件,并通过酸沉-醇沉、超滤和透析对其进行了纯化。结果表明,DNS微孔法测定α-AI活性的最佳条件为:测定波长为482 nm、DNS用量为80μL、反应时间为15 min、静置时间为10 min;α-AI提取的最优工艺为:提取温度为45℃、提取时间为2.5 h、提取p H为6.6、液料比为7.5;经纯化后得到了较纯的、活性较高的α-AI,其IC50为0.89±0.04 mg/mL,提取率为(1.92±0.23)%。该提取纯化工艺可为α-AI的低成本工业化生产提供一定的指导意义。
In order to develop a simple, effective and low-cost extraction and purification process of α-amylase inhibitor (α-AI) and explore the optimal conditions for its activity test, white kidney beans were used as raw materials to extract α-AI by water extraction in this study. After studying the conditions for the determination of α-AI activity by 3,5-dinitrosalicylic acid (DNS) micropore method, the optimal process conditions for α-AI extraction were determined by orthogonal test on the basis of single factor and the α-AI crude extract was purified by acid precipitation-alcohol precipitation, ultrafiltration and dialysis. The results indicated that the optimal conditions for the determination of α-AI activity were: determination wavelength of 482 nm, DNS dosage of 80 μL, reaction time of 15 min, and the rest time was 10 min. And the optimum process for α-AI extraction were: Extraction temperature of 45 ℃, extraction time of 2.5 h, extraction pH of 6.6 and the liquid-solid ratio was 7.5. Moreover, the α-AI with high purity and activity was obtained after purification, which with an IC50 of 0.89±0.04 mg/mL and a yield of 1.92±0.23%. The extraction and purification process may be can provide some guiding significance for the low-cost industrial production of α-AI.
作者
钟颖颖
何绮怡
冯俊超
朱秋雁
张春国
赵肃清
ZHONG Ying-ying;HE Qi-yi;FENG Jun-chao;ZHU Qiu-yan;ZHANG Chun-guo;ZHAO Su-qing(School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510006, China)
出处
《现代食品科技》
EI
CAS
北大核心
2019年第8期254-260,225,共8页
Modern Food Science and Technology
基金
广东省自然科学基金资助项目(S2013030013338)
关键词
白芸豆
Α-淀粉酶抑制剂
提取
纯化
3
5-二硝基水杨酸微孔法
white kidney bean
α-amylase inhibitor
extraction
purification
3,5-dinitrosalicylic acid micropore method
作者简介
钟颖颖(1995-),女,研究生,研究方向:生物制药;通讯作者:赵肃清(1969-),男,教授,博导,研究方向:天然产物分离提取,生物制药及免疫分析.
