產(chǎn)紫杉醇菌種的改良及若干種前體物質(zhì)作用的研究.pdf

1、江南大學碩士學位論文產(chǎn)紫杉醇菌種的改良及若干種前體物質(zhì)作用的研究姓名：代文亮申請學位級別：碩士專業(yè)：發(fā)酵工程指導教師：陶文沂20080401AbstractAbstractTaxol，ahighlyderivatizedditerpenoid，wasfirstlyfoundfromsomespeciesofTaxusDHetoitsuniqueanticancermechanismandtheremarkablecurativeeffe

2、ctforvariousmalignanttumors，ithasbeenthefocustowhichpeoplepayattentionallthetimeTaxolproductionbyplantendophyticfungifermentationhasbeentheresearchhotspotinrecentyearsItisimportanttoimprovetheTaxolproductionbyfungiinord9

3、rtomeettheproductionrequirementBreedingofhighyieldstrainsandoptimalcultivationconditionswillbetheeffectivewaystoincreasetheyieldofTax01ThefurtherresearchbasedtheFusariummairefK178from虹1eprotoplastmutationwasinvestigatedb

4、yoptimizationofbiosynthesisprecursorbreedingmethodandfermentationtechnologytoincreaseTaxolyieldThemainmethodsandresultswereasfollows：1TheconcentrationofprecursorsfortheproductionofTaxolwithFusariummairefK178wasoptimizedb

5、yresponsesurfacemethodologyFirstlyPlackett—BurmandesignwasundertakentoevaluatetheeffectsofeightfactorsWithstatisticregressionanalysis，thesignificantfactorsaffectingTaxolproductionweredeterminedasfollows：phenylalanine，sod

6、iumacetateandsodiumbenzoateBoxBehnkendesignwasusedtooptimizethethreecriticalinternalfactorsmentionedaboveandwefoundout也eoptimumconcentrationlevelsandtherelationshipsamongthesefactorsByquadraticregressionmodelequation、析mD

7、esign—Expertstatisticmethods，theoptimalconcentrationofthevariablesweredeterminedas：phenylalanine28mg／L，sodiumacetate33g幾，sodiumbenzoate318mgmUndersuchconditions，theTaxolproductionwasincreasedto2426pg／L，whichwas156％higher

8、t11啦themaximumvalueinthesinglefactortestsTheexperimentvaluesundertheoptimalconditionsagreed謝t11也epredictedvalueswhichindicatedthatthemodelwasproperandeffectiveAndthemechanismoftheprecursorssynthesisofTaxolwerealsostudied

9、2SporesofTax01producingfungus只裕ariummaireiK178weretreatedwithUVandmtritethemutantswereselectedintheplatewith40pg／mLnystatinTheresultsshowedthat5mutantsexhibitedanincreaseinTaxolproductionamongtheobtained3lnystatinresista

10、ntmutantsTheTaxolproductionofmutantF協(xié)ariummaireiUH23was259。8“g／L，whichwas178％more也anthatoftheStartingstrain3飛hefermentationconditionsofendophyticfungusFuSariummairemH23wereinvestigatedforTaxolproductionTheresultsshowedth

11、ataninoculationvolumeof5％intothe60mL／250mLErlenmeyerflaskattheconditionof26℃andinitialpH70couldreachthemaximumTaxolproductionafterincubationof12daysGlucoseandammoniulnnitratewereselectedasthebestcarbonandnitrogensourcein

12、thefurtherstudyTheresultsshowedthattheoptimalconcentrationswere(g／L)：glucose60，NH4N036，MgS0403，KHaP0405，ZnS041x10。j，Cu(N03)2110一，F(xiàn)eCl3210一，VBx510一，phenylalanine2810一，sodiumbenzoate318X10’1，sodiumacetate33，LTyrosine5X10一T

13、heTaxolyieldreached3059pg／LundertheoptimizedconditionsTheexpressionofnitratenitrogenandammoniumnitrogenaresynergisticeffectsonthebiosynthesisofTax01Undertheconditionofaddingsucroselessthanorequalto10g／Lcarlpromotethecell