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1、现代食品科技ModernFoodScienceandTechnology2016,Vol.32,No.12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朱娟娟,尹忠平,陈继光,上官新晨,彭大勇,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)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0.05U/m
2、L;底物浓度范围为0.05~1mM;反应温度为37℃;反应时间为6min。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-BurkPlots、Eadie-HofsteePlots、Hanes-WolffPlots、Eisenthal-Cornish-BowdenDirectPlots、Non-linearRegressionAnalysis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
3、存在一定的差异,Non-linear-RegressionAnalysis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-RegressionAnalysis-6法计算,该模型中酶促反应的Vmax为3.91×10mmol/min,Km为0.12mM,阿卡波糖的Ki为90μM。关键词:α-葡萄糖苷酶抑制剂;模型;动力学文章篇号:1673-9078(2016)12-164-170DOI:10.13982/j.mfst.1673-9078.2016.12.026AModelforS
4、creeningTraceAmountsofα-GlucosidaseInhibitorsandMethodsforJudgingInhibitionTypeZHUJuan-juan,YINZhong-ping,CHENJi-guang,SHANGGUANXin-chen,PENGDa-yong,JIANGYan(JiangxiKeyLaboratoryofNaturalProductandFunctionalFood,CollegeofFoodScienceandEngineering,JiangxiAg
5、riculturalUniversity,Nanchang,330045,China)Abstract:Alpha-glucosidaseinhibitorscaninhibitcarbohydratehydrolysisandarecommonlyusedtodecreasepostprandialbloodglucoseinindividualswithhyperglycemia.Amodelfortherapidscreeningoftraceamountsofalpha-glucosidaseinh
6、ibitorswasestablishedbasedonthealpha-glucosidase-pNPGinvitroreactionsystem.Theoptimizedparametersofthismodelwereasfollows:theenzymeconcentration,substrateconcentration,reactiontemperature,andreactiontimewere0.05U/mL,0.05~1mM,37℃,and6min,respectively.Theinh
7、ibitoryeffectofacarbosewasdeterminedusingthismodel,andtheenzymaticreactionkineticsdatawereanalyzedusingfivemethods(Lineweaver-Burkplots,Eadie-Hofsteeplots,Hanes-Wolffplots,Eisenthal-Cornish-Bowdendirectplots,andNon-linearregressionanalysis).Dataprocessinga
8、ndcomparisonofresultsshowedthatthesefivemethodshaddistinctcharacteristicsastheKi,Km,andVmaxcalculatedbythesefivemethodsdifferedslightly.Non-linearRegressionAnalysiswasmoreconvenient,reasonable,andreliable,and