南方紅豆杉細胞中紫杉烷生物合成途徑及其誘導(dǎo)調(diào)控研究.pdf

1、天津大學(xué)博士學(xué)位論文南方紅豆杉細胞中紫杉烷生物合成途徑及其誘導(dǎo)調(diào)控研究姓名：王艷東申請學(xué)位級別：博士專業(yè)：生物化工指導(dǎo)教師：元英進20021201ABSTRACTThemetabolismpathwaysofisopentenylpyrophosphate(IPP)andtheimportantfunctionofthecgocllromep450monooxygenasesfortaxanebiosynthesesinsusDensi

2、onculturesofTaxuschinensisvarmairei(T)wereinvestigatedusingsomemetabolicinhibitorsThedetailstudiesweredonefromtheaspectsasfollowsAtfirst，somemetabolicinlfibitors(mevastatin，chlorocholinechloride，sodiumpyrophosphateandD，L

3、glyceraldehyde)wereaddedintosuspensionculturesofTaxuscellsinordertounderstandthemetabolismpathwaysofIPPfortaxanebiosynthesesTheresultsforthefirsttimeshowthatIPPmetabolismpathwaysweredifferentatthedifierentgrowthphasesofT

4、axuscellsThenonmevalonatepmhwaywasinvolvedinIPPbiosynthesisfortaxanebiosynthesesattheearlygrowthphaserO14day)ofTaxuscellsAtthelategrowthphase(afterday14)，themevalonatepathwayinthecytoplasmandIPPtranslocationwereinvolvedi

5、ntaxanebiosynthesesTheactionmechanismofmethyljasmonate(MJ)onTaxuscellsmetabolismwassystemicallyinvestigatedbyanalysisofcellmorphologyandthesolutionofparametersininductionmodelandSOonMoreoverbyusingthemethodsofmetabolicin

6、hibition，theinductionofMJonIPPmetabolismwasstudiedbyanalyzingtheinhibitionratioandinductionratioTheresultssuggestthatMJstronglyinducedthenonmevalonatepathwayin10一DAB(10一deacetylbaccatinIII)andbaccatinIIIbiosynthesesatthe

7、lategrowthphaseInaddition，MJinducedthemevalonatepathwayintaxolbiosynthesisMJstimulatedIPPtranslocationfromthecytoplasmtotheplastidsinbaccatinIIIbiosynthesisatthelategrowthphaseFurthermore，itisfoundthatMJdidnotinduceIPPtr

8、anslocationfor10DABandtaxolbiosynthesisUsingthemethodofinhibitorblockingmetabolicpathways，themetabolismpathwaysofIPPundertreatmentwithsalicylicacid(SA)wereinvestigatedcomparedwiththatofMJTheresultsshowthatSAobviouslyindu

9、cedthenon—mevalonatepathwayintheplastidsfor10一DAB，taxolandcephalomanninebiosynthesesandIPPtranslocationfortaxolbiosynthesisatthelategrowthphaseofTaxuscellsTheinductionofMJandSAoncNocllromeP450monooxygenasesintaxanebiosyn

10、theseswasinvestigatedbyaddingthespecificP450monooxygenaseinhibitor，1一aminobenzotriazole(ABT)，intosuspensionculturesofTaxuscellsTheresultsindicatethattheinductionofMJonP450enzymeswasstrongerattheearlygrowthphaseofTaxuscel