Separation,PurificationandCharacterizationofThreeEndo-polygalacturonasesfromaNewlyIsolatedPenicillumoxalicum

Threeendo-polygalacturonases(endoPGs)fromanewlyisolatedPenicillumoxalicum(CGMCC0907)capableofutilizingwastebiomassasgrowthsubstratewereseparatedandpurifiedtohomogeneitybyultra-filtration,affinityadsorptionchromatography,CM-cellulosecolumnchromatography,andSephadexG-100gelfiltrationchromatographywiththeoverallyieldof64.5%fromthecrudeenzyme.ThespecificactivitiesandrecoveryratesofendoPG-1,endoPG-2andendoPG-3were1120U/mgand21.6%,1350U/mgand25.9%,and1560U/mgand17.0%,respectively.ThethreepurifiedendoPGshadaclosemolecularweightto41kDaasestimatedbySDS-PAGE.TheoptimumtemperatureandpHforthefunctionofthemwere65℃and5.0,55℃and5.0,50℃and5.5,respectively.TheirpIandKmvalueswere5.9and0.78mg/mL,6.0and1.2mg/mL,and6.1and2.0mg/mL,respectively.