朱娟娟,尹忠平,陈继光,上官新晨,彭大勇,蒋艳.微量α﹣葡萄糖苷酶抑制剂筛选模型及抑制类型的判断方法[J].,2016,32(12):164-170.
微量α﹣葡萄糖苷酶抑制剂筛选模型及抑制类型的判断方法
A Model for Screening Trace Amounts of α-Glucosidase Inhibitors and Methods for Judging Inhibition Type
投稿时间:2015-12-24  
DOI:10.13982/j.mfst.1673-9078.2016.12.026
中文关键词:  α-葡萄糖苷酶抑制剂  模型  动力学
英文关键词:alpha-glucosidase inhibitor  model  kinetics
作者简介:朱娟娟(1989-),女,硕士研究生,研究方向:天然产物与功能性食品 通讯作者:尹忠平(1971-),男,博士,副教授,研究方向:天然产物与功能性食品
基金项目:国家自然科学基金资助项目(31460436,31260368);江西省自然科学基金项目(20132BAB204004)
作者单位
朱娟娟 (江西农业大学食品科学与工程学院,天然产物与功能食品重点实验室,江西南昌 330045) 
尹忠平 (江西农业大学食品科学与工程学院,天然产物与功能食品重点实验室,江西南昌 330045) 
陈继光 (江西农业大学食品科学与工程学院,天然产物与功能食品重点实验室,江西南昌 330045) 
上官新晨 (江西农业大学食品科学与工程学院,天然产物与功能食品重点实验室,江西南昌 330045) 
彭大勇 (江西农业大学食品科学与工程学院,天然产物与功能食品重点实验室,江西南昌 330045) 
蒋艳 (江西农业大学食品科学与工程学院,天然产物与功能食品重点实验室,江西南昌 330045) 
AuthorInstitution
ZHU Juan-juan (Jiangxi Key Laboratory of Natural Product and Functional Food, College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang, 330045, China) 
YIN Zhong-ping (Jiangxi Key Laboratory of Natural Product and Functional Food, College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang, 330045, China) 
CHEN Ji-guang (Jiangxi Key Laboratory of Natural Product and Functional Food, College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang, 330045, China) 
SHANG GUAN Xin-chen (Jiangxi Key Laboratory of Natural Product and Functional Food, College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang, 330045, China) 
PENG Da-yong (Jiangxi Key Laboratory of Natural Product and Functional Food, College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang, 330045, China) 
JIANG Yan (Jiangxi Key Laboratory of Natural Product and Functional Food, College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang, 330045, China) 
摘要点击次数: 798
全文下载次数: 1766
中文摘要:
      α-葡萄糖苷酶抑制剂能抑制碳水化合物水解,是高血糖人群降低餐后血糖的常用物质。本文基于α-葡萄糖苷酶-PNPG体外反应体系,建立了微量、快速的α-葡萄糖苷酶抑制剂筛选模型,该模型的主要参数如下:酶浓度为0.05 U/mL;底物浓度范围为0.05~1 mM;反应温度为37 ℃;反应时间为6 min。以该模型检测了阿卡波糖对α-葡萄糖苷酶的抑制作用,并采用Lineweaver-Burk Plots、Eadie-Hofstee Plots、Hanes-Wolff Plots、Eisenthal-Cornish-Bowden Direct Plots、Non-linear Regression Analysis五种方法对该酶促反应的动力学数据进行了详细的分析。通过对数据处理的过程和结果的比较发现,该五种方法各有特点,各法所获得的Vmax、Km和Ki存在一定的差异,Non-linear-Regression Analysis法更加简便、合理及可靠,是酶动力学数据处理的首选方法。采用Non-linear-Regression Analysis法计算,该模型中酶促反应的Vmax为3.91×10-6 mmol/min,Km为0.12 mM,阿卡波糖的Ki为90 μM。
英文摘要:
      Alpha-glucosidase inhibitors can inhibit carbohydrate hydrolysis and are commonly used to decrease postprandial blood glucose in individuals with hyperglycemia. A model for the rapid screening of trace amounts of alpha-glucosidase inhibitors was established based on the alpha-glucosidase-pNPG in vitro reaction system. The optimized parameters of this model were as follows: the enzyme concentration, substrate concentration, reaction temperature, and reaction time were 0.05 U/mL, 0.05~1 mM, 37 ℃, and 6 min, respectively. The inhibitory effect of acarbose was determined using this model, and the enzymatic reaction kinetics data were analyzed using five methods (Lineweaver-Burk plots, Eadie-Hofstee plots, Hanes-Wolff plots, Eisenthal-Cornish-Bowden direct plots, and Non-linear regression analysis). Data processing and comparison of results showed that these five methods had distinct characteristics as the Ki, Km, and Vmax calculated by these five methods differed slightly. Non-linear Regression Analysis was more convenient, reasonable, and reliable, and therefore was the first choice for processing the data of enzyme kinetics. By this model, the Vmax (?10-6 mmol/min) and Km (mM) of enzymatic reaction were calculated to be 3.91 and 0.12, respectively and the Ki (μM) of acarbose was 90.
查看全文  查看/发表评论  下载PDF阅读器
关闭