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1、現(xiàn)代食品科技ModernFoodScienceandTechnology2016,Vol.32,No.12微量α﹣葡萄糖苷酶抑制劑篩選模型及抑制類型的判斷方法朱娟娟,尹忠平,陳繼光,上官新晨,彭大勇,蔣艷(江西農(nóng)業(yè)大學食品科學與工程學院,天然產(chǎn)物與功能食品重點實驗室,江西南昌330045)摘要:α-葡萄糖苷酶抑制劑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質(zhì)。本文基于α-葡萄糖苷酶-PNPG體外反應體系,建立了微量、快速的α-葡萄糖苷酶抑制劑篩選模型,該模型的主要參數(shù)如下:酶濃度為0.05U/m
2、L;底物濃度范圍為0.05~1mM;反應溫度為37℃;反應時間為6min。以該模型檢測了阿卡波糖對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五種方法對該酶促反應的動力學數(shù)據(jù)進行了詳細的分析。通過對數(shù)據(jù)處理的過程和結果的比較發(fā)現(xiàn),該五種方法各有特點,各法所獲得的Vmax、Km和Ki
3、存在一定的差異,Non-linear-RegressionAnalysis法更加簡便、合理及可靠,是酶動力學數(shù)據(jù)處理的首選方法。采用Non-linear-RegressionAnalysis-6法計算,該模型中酶促反應的Vmax為3.91×10mmol/min,Km為0.12mM,阿卡波糖的Ki為90μM。關鍵詞:α-葡萄糖苷酶抑制劑;模型;動力學文章篇號: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