基因工程菌全细胞催化制备葡甘低聚糖

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1、万方数据ABSTRACTGlucomannan—oligosaccharidesarewater-solublepolysaccharidepolymers，asfunctionaloligosaccharides，whichcanimproveanimalintestinalmicroflora，improveimmunefunctionandoxidationresistance，widelyusedinfood，medicineandfeedindustry．Theirspecialphysio

2、logicalfunctionsandbroadapplicationprospectsgreatlyarousedtheinterestofscholars．Theglucomanno-oligosaccharideswerepreparedwithrecombinantengineeredbacteriacontainingbetamannasegenebywhole-cellcatalysis；TheseenzymatichydrolysatesweredetectedbyTLCandHPLC，

3、bleachedandprocessedtOremovemonosaccharides．Theglucomanno-oligosaccharideswerepreparedwiththeRecombinantengineeredbacteriaRossate—pET32a—CH3containingbetamannasegenebywhole-cellcatalysis．Bysinglefactoroptimizationanalysis，theinfluencesofdifferentdigesti

4、ontemperature，digestiontime，pH，substrateconcen仃ationandenzymeamountontherateofhydrolysiswereexplored．Theresultsshowedthatenzymatichydrolysisreactionofoptimumtemperatureof50℃，thebesttimeof12h，thebestpHvalueof7．0，thebestsubstrateconcentrationwas3％，thebest

5、enzymeamountwas1000U．Becausetheinfluenceoftimeandenzymeonhydrolysisratewasnotobvious，reactiontemperature(A)，substrateconcentration(B)，pH(C)wereselectedasthethreevariables，hydrolysisrateYasresponsevalue．AccordingtotheprincipleofBox-Behnken，testplanwerede

6、signedbytheDesign·Expertsoftware．Finally，testresultsdeterminestheoptimumcondition：reactiontemperaturewas50．6℃，thesubstrateconcentrationwas3．4％，pHvalueof6．8，makinganeventualkonjacpowderhydrolysisratereached41．34％，inconformitywiththepredictivevalueof41．54

7、1％．Becauseofthethinlayerchromatography(TLC)withlowcost，fastdetection，theadvantagesofsimpleoperation，therefore，glucomanno-oligosaccharideswerepreparedandenzymeproductsweredetectedbythinlayerchromatography．Enzymolysisliquidandoligosaccharidesstandardconta

8、iningmannose，mannobiosemanninotriose，mannotetrosewerespotonSilica60silicagelplate，toanalyzebythinlayerchromatography．Theanalysisresultsshowedthemaincomponentsofenzymatichydrolysisproductweremannobiosemanninotriose，mannotetrose，di