蛋白质水解度的简易测定方法

a:10083464(2002)02011303袁斌1,吕桂善2,刘小玲2(1,518019;2,530004):,pHSTAT,,,,:;;pHSTAT:TQ936.1:AThesimplemethodofdeterminingforthedegreeofhydrolysisofproteinsYUANGBin1,LUGuishan2,LIUXiaoling2(1ShenzhenChengyeFreezeCorporationLimited,Shenzhen518019,China;2BiotechnologyandSugarEngineeringCollege,GuangxiUniv.,Nanning530004,China)Abstract:Asimpleandworkablemethod,whichwasbasedonpHSTATmethod,fordeter-miningthedegreeofhydrolysisofprotein,wasintroducedinthepresentpaper.Thismethodcanbeusedinlabandfoodindustrywithgoodresultsandlittlereagentsconsumptions.Keywords:protein;degreeofhydrolysis;pHSTATMethod,,,,,,,,,(Degreeofhydrolysis,DH),:DH%=h/htot100%,h(mmol/g),htot,,,,,,htot8.2mmol/g212Vol.21,No.2JournalofGuangxiAgric.andBiol.Science20026June,2002a:20010818:(X0102019):(1973),,,;,,100%;,0-NH2-COOH,,,-NH2-COOHh,(TNBS)[1],,[2],pHSTAT[3],,,;,,[4],TNBS,,[2,4],pHSTATpHSTAT,,,pHSTAT11.1:1kg,6mol/LHClpH4.6,,4000r/min15min,,pH,,,1.2(Trypsin,Gibco,1250),20mmol/L(pH7.10),0.5mol/LNaOH1.31.0000g,25mL20mmol/L(pH7.10),pH7.0,100mg,,50,,,100,5min,,1.4pHSTATpHSTAT[3],pHpH,pH1.5(DH%)pHSTAT[3],,pHSTAT,pH,pH7.0,:,pH7.0,pH,0.5mol/LNaOHpHpH,,[3]:DH%=B(Mb)(1/A)(1/MP)(1/htot)100:BNaOH(mL);MbNaOH(mol/L);pH7.0,50,,1/A2.26;MP(g);htot,,8.22(1),,pHSTAT,11,pHSTAT(2),,,,2,11421pH,,,pHSTAT,,1h,(2),(3)(1624h),pH,pH,,,pHSTAT1Table1ThedeterminationofthedegreeofhydrolysisofcaseinMethodpHBeginningpHvaluepHTerminationpHvalue0.5molL-1NaOH/LLVolumeof0.5molL-1NaOH/%DegreeofhydrolysispHSTATpHSTATmethod7.07.0105014.47Simplewayinthepresentpaper7.06.20104014.33:240minNote:Thetotalhydrolysistimewas240min.2Table2TheparametersofcaseinhydrolysisprocessItemMethodTime/min306090120150180210240pHpHvalueA7.007.007.007.007.007.007.007.00B6.576.346.326.326.326.326.336.34/LLA950.00Volumeof0.5molL-1NaOHB1000.00/%A13.09DegreeofhydrolysisB13.78:ApHSTAT,B30minpH,pH7.0Note:AdenotesthetypicalpHSTATmethod;Bdenotesthesimplewayusedinthepresentpaper.3pHSTAT,,pH,pHpH,pH,pH,pHSTATDH%:[1]ADLERNISSENJ.DeterminationofthedegreeofhydrolysisoffoodproteinhydrolysatesbyTrinitroben-zenesulfonicAcid[J].JAgricFoodChem,1979,27:12561262.[2],.[J].,1994,11:6567.[3]ADLERNISSENJ.EnzymaticHydrolysisofFoodProteins[M].London:ElsevierAppliedSciencePublishers,1986.1214.[4].[J].,2000,25(6):176177.()1152: