2017 7, (1),, 1.3 [12~14], 1 40,,, 3,200 g 1.1 5min 2mg 1 ml, min,, : 10mL15%, 40 min Mark:18~94ku, Thermo, 4 ml3%, : 800U/mg, Bioreagen

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2017 7 Vol.33,No.7 Jul.2017 DOI:10.13652/j.issn.1003-5788.2017.07.

1 Vol.33,No.7 Jul.2017 DOI: /j.issn Analysisofglycosylatedcaseininwholemilkpowder SUN Jing-li LI Lin-qiang ZHU Li-li TANG Lu WU Yuan (, ) (Colegeof Food Engineering and NutritionalScience,Shaanxi Normal University,Xi'an,Shaanxi710119,China) :, groups.thepercentage(46.13±0.46)% ofcaseinextracted ph bytannicacid wassignificantlyhigherthanthatofother methods, SDS-PAGE,.TheresultsofcaseinSDS-PAGEelectrophoresisboth, showedthesame3bandseitherdiferentacidsordiferentmethods, oncaseinextraction,andtheirmolecularweightswere34,24,and,ph4.6 60ku,respectively,andthe34and24ku wereglycosylatedcasein bythemethodofperiodate-schifalkalidyeing.theresultsofmorse 2% (ph3.58) (40.45± experimentshowedthatmilkpowdercaseinexistglycosylatedcasein. 0.66)%, Thecontentoffreeaminoacidfrom wholemilkpowderwassignifi- (46.13±0.46)%, cantlyhigherthanthatfromfreshmilkafterpapainhydrolysis(p< SDS-PAGE, 0.05).Wholemilkpowderhasglycosylatedcasein,theirmolecular 3, 34,24,60ku weightsare34and24kurespectively,whichareeasytohydrolysis., 34,24ku Keywords:wholemilkpowderreconstitutedmilkcaseinglycosy-, lation,, 34,24ku, : Abstract:Thisstudyinvestigatedglycosylatedcaseinin whole milk powderandexploreditsenzymaticproperty.thecaseinextraction weightpercentageweredeterminedbydiferentph,diferentacids, anddiferentmethods.thecaseinwasanalyzedbysds-pageelec- trophoresis,and the glycosylated casein wereidentified via the methodofperiodate-schifalkalidyeingandtheexperimentofmorse.the caseinwashydrolyzedbypapainandtheaminoacidcontentwasdeter- minedaccordingly.results:thepercentageofcaseinextractedunderph 4.6wassignificantlyhigherthanthatofothergroups.The weightpercentageofcaseinextractedfrom aceticacid (ph 3.58)was (40.45±0.66)%,which wassignificantlyhigherthanthatofother [1],, 50%, [2-3] [4], [5] α-1- (AGP) [6] [7-8] [9-11],,, : ( :2016NY-212) 170 :,,,, : (1971 ),,,, lilinq@snnu.edu.cn : , 6

2 2017 7, (1),, 1.3 [12~14], 1 40,,, 3,200 g 1.1 5min 2mg 1 ml, min,, : 10mL15%, 40 min Mark:18~94ku, Thermo, 4 ml3%, : 800U/mg, Bioreagent, 1h,, :, 10μL, 80V 40min,120V : (Tris) N,N,N,N- (TEMED) (SDS) G-250 α- :, [15-17] ( ) :800B, : :PowerPacTM Universal, 10% Bio-rad :KQ-250DB, 1h, 2 ( 0.5h), Schif 1h(Schif : 1g 400 ml, :HY-2, 12mol/L HCl4.2 ml 3.2g,4, :722, 2g, 1min ), (0.5% 1.2 7% 1 1 ),4, [18], : (g/ml), 5 ml,, 1 ml 1% 1% ph(4.6,4.8,5.0,5.4,5.6,5.8) ( 5mL 0.1% NaCl ), (5% 200 g 10min,,, (1) α-, 95%, )2, W = m1 m 100%, (1) : W,% m1,g m,g (g/ml) ( 0.1% NaCl ), 10 ml,, 5 ml, 1mL 0.1% ( 0.1% NaCl, (0.5%,1%,2%,3%,4%,5%), ), 37 6h [19-20] ph,200 g 10min,, 1.5.2, (1) (0.0,1.0,2.0,5.0,10,15,20μg/mL) 1.0 ml mL, 2 mlph5.5 5mL, 10 mlph4.6 ( ) ph4.6 ( 2.5h, G-250 2h,, ( ) SDS-PAGE 1h, 2 ( 0.5h), 1%, 1.5mL 2, ph4.6, 0.1%,, 2% (2g 100 ml, )2 ml 15 min,, ) 0.1% ( ) ( 570nm, (y =0.0319x + ) 5% ( ), ,R 2 =0.9995), 40,,, 7

3 : 1.6 3, SPSS19.0, ±, LSD 5% ph 4.6, (28.38±2.49)%,, [21-22] ph4.6 3 ph, Figure3 Influenceoflacticaciddiferentadditionsonex-, tractionrateofproteinandphvalueofsolution 1 Figure1 Influenceofaceticacid-sodiumacetatebufer solutiononproteinextractionrate 2.2 2~5, 2%, (40.45±0.66)%, 2%, (39.31±2.76)%, 3%, (37.44±1.04)%, 4%, (31.74±4.20)%,, > > > ph 5 ph > > >, ph Figure5 Influenceofphosphateaciddiferentadditionson,,, extractionrateofproteinandphvalueofsolution [23], 4 ph Figure4 Influenceofhdrochloricaciddiferentadditionson extractionrateofproteinandphvalueofsolution, 2.3 6, (46.13± 0.36)%,, (P< 0.05)[ 3 (P>0.05)], [24~25], 2 ph,, Figure2 Influenceofaceticaciddiferentadditionsonex- tractionrateofproteinandphvalueofsolution,, 8

AnalysisofSDS-PAGEoncaseinfromreconstitu-

4 SDS-PAGE ( 9b), 24,34ku,, 7, 3 9a, 60ku

4 ( 9a), a. G-250 b. 1~ Figure9 AnalysisofSDS-PAGEoncaseinfromreconstitu- Figure6 Influenceofdiferentmethodsonprotein tedmilktreatedbydiferentdyeingmethods extractionrate 24,34,60ku SDS-PAGE ( 9b), 24,34ku,, 7, 3 9a, 60ku Mark, 24,34,60ku,, [29~32] [26~28] 2.6, 10 8,, 3, Mark,, 24,34,60ku,, 2.5,, Figure10 1~4., 7 ph4.6, 2 [33~36] Figure7 AnalysisofSDS-PAGEoncaseinfromreconstituted milktreatedbydiferentacidsatph4.6, [37-38],, 10 Morseexperimentofcasein 2.7, ( ) 1~ Figure8 AnalysisofSDS-PAGEoncaseinfromreconstitu- tedmilktreatedbydiferentextractionmethods 11 Figure11 Influenceofglycosylationonhydrolysisdegree ofcasein 9

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