利用里氏木霉生物转化制备黄姜薯蓣皂甙元的清洁新工艺

46420107ActaScientiarumNaturaliumUniversitatisPekinensisVol.46No.4July2010“”2006BAB04A142009-06-012009-08-1712111.1008712.610041[e-mail]。。98.7%98.0%99.4%COD100%SO2－4100%。24-2-。17β--5--3-。。X506AGreenExtractionProcessofDiosgeninfromDioscoreazingiberensisTuberswithTrichodermareeseiZHUYuling1HUANGWen2LIUWei1NIJinren11.KeyLaboratoryofWaterandSedimentSciencesMinistryofEducationDepartmentofEnvironmentalEngineeringPekingUniversityBeijing1008712.StateKeyLaboratoryofBiotherapyofHumanDiseasesCancerCenterWestChinaHospitalWestChinaMedicalSchoolSichuanUniversityChengdu610041[e-mail]ofresourceefficiencyandcleanerproductionagreenextractionprocessofdiosgeninfromDioscoreazingiberensistuberswasproposed.ThismethodcontainedofexcludingstarchfromrawmaterialandbioconvertingthesaponinsintheresiduebyTrichodermareesei.Comparedtoacidhydrolysis98.7%ofdiosgenincouldbeextractedfromplanttubersthroughthenewprocesswith98.0%ofstarchrecoveredintheformofsugar.Consequently99.4%ofCOD100%ofSO2－4and100%ofacidwerereducedinprocessingwastewater.Inadditionacidhydrolysisby-productsweregreatlylimitede.g.twoprioritypollutantsBis2-ethylhexylphthalateandDi-n-butylphthalatewhicharespecifiedbyUnitedStatesEnvironmentalProtectionAgencytypicallydetectedinDioscoreazingiberensiswastewaterfromconventionalacidhydrolysisprocessdisappearedautomaticallyintheproposedcleanerprocess.KeywordsdiosgeninsaponinsTrichodermareeseiDioscoreazingiberensis、、1-2。60%3-4。166465-6。D.zingiberensis。26-3β-1。7-8。9-10。COD50%～70%CODSO2－4H+。、。。Huang11+β-+95%12Aspergillussp.304295%。。30%～50%13-16。90%。。Huang1112。。11.1。T.reesei。50%4℃PDA。。α-2000IU/g10000IU/g。1.260℃500g3000mL5.0L1hpH6.510g70℃1hpH4.015g60℃8h。4000rpm15min60℃。100。1.31.3.130g/L3g/LNaNO31g/LK2HPO40.5g/LMgSO40.01g/LFeSO40.1g/LTween-80。w/V9.77%2.67%0.29%K2HPO40.73%Tween80pH5.8。1.3.275mL0.01%Tween80100mL250mL。30℃200rpm。10mL5±1g/L100mL500mL30℃200rpm168h。12h10000rpm4℃15min。αβ-。1Fig.1Hydrolyzingpathwayofsaponinstodiosgenin2664300mL3.0L5.0L0.8m3/m3·min300rpm30℃156h。10000rpm4℃15min。。1.41.4.117。1.4.2DNS18。1.4.3β-1mL50℃10min50℃10minpNPG-β-D-0.5mmol/LpH6.00.02mol/L50℃250rpm10min。1mL1mol/LNa2CO35mL。420nm。1minpNPG1μmol1IU/mL。1.4.430min。HPLC。HPLC210nm10μLZOR-BAX5B-C1815cm×4.6mm5μm40℃∶=1∶9。1mL/min。Y=Dm－D0Da－D0×100%。Y%DmDaD0。31mol/LH2SO4121℃3h4000rpm15min。CHCl3。1.4.5CODTOC/SO2－4BaCl219。1mLGC-MS。DB-3530.0m×0.25mm×0.25μm280℃40℃3min5℃/min100℃5min5℃/min150℃5min5℃/min290℃5min1mL/min1.0μL。EM1494V。55～550m/z。NIST0.5。22.1137.5%3.03%。2.11%9.25%。1g0.36g。98.0%9.90%。1Table1ChemicalanalysisofD.zingiberensistubersandpretreatedD.zingiberensistubers%8.37±0.157.91±0.1037.5±0.982.11±0.875.74±0.161.08±0.0745.3±3.7550.9±5.273.03±0.219.25±0.27。20。。2.22.2.1β-246。2。T.reesei。48。843664696β-。15690.6%。T.reesei156h90%。3CHCl3HPLC-UV210nm。3abc、CHCl3。9.55min3d。。2、、β-Fig.2Timecourseofreducingsugarbiomassβ-glucosidasediosgenininshakeflaskcultivationofpretreatedDioscoreazingiberensistubersbyT.reesei3abcdCHCl3HPLC-UV210nmFig.3TheHPLC-UV210nmchromatogramsoftheCHCl3extractfromuntreatedamicrobialhydrolyzedbandacidichydrolyzedcD.zingiberensisresiduedrepresentstheHPLC-UV210nmchromatogramofstandarddiosgenin46642.2.2300rpm0.8m3/m3·min。156h90.2%。2.3100g2.97g0.41gCOD100g3.01g65.8gCOD10.2gSO2－40.18gH+。99.4%COD100%SO2－4100%。2GC/MSTable2OrganiccompoundsinacidprocessingandnewprocesswastewateranalyzedbyGC/MS√－√－√－√－*√－√－√－√－1-√－√－√－1--√－√－√－√－√－-35-11--4-√－√－-2-*√－25--14-√－√－510--2378--1H6H-12-a1′2′-d√－17β--5--3-√√－√√－。*EPA。、、COD。。GC/MS2。239142。-2-EPA21。17β--5--3-。GC/MS。31。2T.reesei90%。98.7%98.0%99.4%COD100%SO2－4100%。24、-2-。3。。1..198314-1112..1994656646193-973..200214389-904..200528395-975..200839443-466..2008392165-1677ZhangCaixiangWangYanxinYangZhihuaetal.Chlorineemissionanddechlorinationinco-firingcoalandtheresiduefromhydrochloricacidhydrolysisofDiscoreazingiberensis.Fuel20068514/152034-20408..2008311136-1389ZhaoHuazhangChengPengZhaoBinetal.Yellowgingerprocessingwastewatertreatmentbyahybridbiologicalprocess.ProcessBiochem200843121427-143110..200438456-5811HuangWenZhaoHuazhangNiJinrenetal.ThebestutilizationofD.zingiberensisC.H.Wrightbyaneco-friendlyprocess.BioresourTechno200899157407-741112.3.200928294-9713..2007308905-91014..2007288269-27315QiShanshanDongYueshengZhaoYangkunetal.IdentificationandquantificationofsteroidalsaponinsandaglyconeduringbiotransformationofDioscoreazingiberensisbyAspergillusoryzae.JBiotechnol2008136S1356-40116QiShanshanDongYueshengZhaoYangkunetal.QualitativeandquantitativeanalysisofmicrobialtransformationofsteroidalsaponinsinDioscoreazingiberensis.Chromatographia2009699/10865-87017ZhangQLoCMJuLK.FactorsaffectingfoamingbehaviorincellulasefermentationbyTrichodermareeseiRutC-30.BioresourTechnol2007984753-76018MillerGL.Useofdinitrosalicylicacidreagentfordeterminationofreducingsugar.AnalChem195931426-42819DongLihongLiWenfeng.Rapiddeterminationofsulphateinwater.IndWaterTr200020334-3520WangYanxinLiuHuiBaoJianguoetal.Thesaccharification-membraneretrieval-hydrolysisSMRHprocessanovelapproachforclean