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ProductionIofIsesquiterpenesIbyIbiocatalyticItransformationIofIsyntheticIfarnesylIdiphosphate

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Acknowledgements

Literature

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TerpenesvarevavlargevandvstructuallyvdiversevclassvofvsecondaryvmetabolitesEvwhichvarevmainlyvproducedvbyvplantsvasvthevmajorvcomponentsvinvresinsvandvessentialvoilsPvZspeciallyvmonoxvandvsesquiterpenesvexhibitvoftenvavstrongvsmellv andvarev thusv fequentlyvusedvasv fragrancevcompoundsv invperfumesvandvvariousv scentedvhouseholdvgoodsPvThesevdaysvthevmajorityvofvterpenevproductionvisvstillvbasedvonvthevisolationvfromvplantvderivedvrawvmaterialsPvDnvincreasingvconsumervdemandEvcombinedvwithvcropvfailuresvandvpoliticalvrisksvinvthevproducingvcountriesvisvleadingvtov higherv pricesv forv highv valuablev essentialv oilsv suchv asv patchouliv oilPv Thereforev newEv climaxindipendentv andvreliablevprocessesvforvthevproductionvofvterpenesvarevrequiredPvWevherebyvpresentvavbioprocessvbasedvonvthevlargexscalev productionvofv farnesylv diphosphatev LXPPbv asv substratev forv av recombinantlyv expressedvpatchoulolv synthasevLseevalsovpostervDvqjbv[°]P

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Thevbiocatalyticvproductionvofv sesquiterpenesvwasv performedv inv av twoxphasev stirredxtankxreactorPvDnvaqueousvMOPSOvbuffervcontainingvvthev patchoulolv synthaseEv XPPv andv MgOlKv wasvusedvforvthevenzymaticvconversionPv–nvordervtovisolatev thev veryv hydrophobicv sesquiterpenesvthev solutionv wasv coveredv withv av layerv ofvorganicv solventv [(]Pv Xorv thatv purposev severalvsolventsv havev beenv testedv inv regardv tovextractionv propertiesEv enzymev compatibilityvandv safev andv easyv handlingPv isoxoctanev andvhexamethyldisiloxanev L9MMSObv havev beenvfoundvtovbevoptimalvsolventsvinvthisvprocessP

Dfterv thev reactionEv thev organicv layerv wasvseparatedv inv av settlerPv –nv orderv tov ensurevcompletleyv productv recoveryEv thev aqueousvphasevwasv extractedvwithv organicv solventPvMuev tov thev highv proteinv contentv thevextractionvwasvsupportedvbyvcentrifugationvtov obtainv phasev separationPv SubsequentlyEvthev solventv wasv removedv carefullyv undervvacuumv resultingv inv av yellowv oilPv ThevsuccessfulvformationvofvsesquiterpenesvwasvconfirmedvbyvJOvandvJOxMSPvXorvthevexactvproductvquantificationvavmethodvbasedvonvShimadzuv JOxKN°Nv usingv av ZRxWDXplusvcolumnvwasvdevelopedPv

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ThisvworkvisvfundedvbyvthevZuropeanvRegionalvMevelopmentvXundvLZXRZb2v–nnovationvNetworkv“Refinmentvofvplantvresources“vLZWv[x[N°(N])NbWevwouldvlikevtovthankvProfPvRergervandvMrPv“ringsvL–nstitutevofvXoodvOhemistryEvLeibnizvUniversityv9anoverbvforvassistancevwithvterpenevanalyticsPWevwouldvlikevtovthankvSymrisevDJvforvthevprovisionvofvsubstratesPv

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ConclusionIandIOutlook

phase boundary