抗生素的产生精讲.ppt

第九章：抗生素的生产,抗生素的生产,概述抗生素的分类抗生素的应用半合成抗生素青霉素的生产和提取工艺,抗生素的生产,定义：是生物在其生命过程中产生的，并在低浓度下有选择性地抑制或杀灭其它微生物或肿瘤细胞的有机物质。MicrobialProducts,effectiveatlowconcentrations,whichhavetheabilitytokillorinhibitthegrowthofcertainmicro-organisms.History:19世纪70年代，法国Pasteur,发现有些微生物可以抑制炭疽杆菌，提出了利用微生物抑制另一种微生物的现象来治疗一些感染性疾病。1928年，,概述,In1928ScottishbacteriologistAlexanderFlemingreportedtheantibacterialactionofculturesofaPenicilliumspeciesHeobservedthatPenicilliumnotatumkilledsomeofhisStaph.aureusculturedishesDiscoverybyaccident?CulturedishescontaminatedwhenleftoutfortwoweekswhileFlemingwasonholiday,insteadofbeingincubated,FlemingnoticedthatthesubstancesecretedwasaninhibitorofmanybacterialspeciesHepublishedapaperontheactiveingredientwhichhecalledpenicillin.Thiswasthefirstdemonstrationthatasubstanceproducedbymicro-orgscouldinhibitorkillothermicrobesFlemingcouldnotisolatepenicillin.10yrslaterscientistsFloreyandChainsuccessfullyisolatedpenicillin.1943-1945，scale-upproductionofpencillium.By1945,enoughpenicillinwasproducedtotreat7millionperyear.,DiscovererofpenicillinBornintoafarmingfamilyinScotland,movedtoLondonat13AwardedScholarshiptostudymedicine,AlexanderFleming,ZoneofInhibition,Thispictureillustratestheeffectofpenicillinonafungus.Notetheclearringofinhibition.ItwasthisobservationwhichinitiatedFlemingsfurtherstudies.,HowardFlorey,Australian-bornStudiedScience&MedicineinAdelaideAwardedScholarshipatOxfordUniversityReturnedtoEnglandtocompleteaPhD,ErnstChain,OfRussian-GermandescentGraduatedinChemistryandPhysiologyLeftBerlinwhenNaziscametopowerHiredbyFloreytoworkonpenicillinproject,到目前为止，有8000多种抗生素，2/3是由放线菌（strepomyces）产生的，并通过化学结构的改造，总共有3万多种半合成抗生素，它们的分子量为100-1200，普遍应用的包括半合成和盐类的抗生素有350多种，其中以青霉素、头孢霉素、四环素类、氨基糖苷类和大环内脂类,主要抗生素的使用量（Harvey,1998),SomeClinicallyImportantAntibiotics,抗生素的分类,根据抗生素的来源分放线菌产生的，占目前发现的抗生素的2/3。真菌产生的，主要包括青霉菌和头孢菌素。细菌产生的，多粘杆菌，枯草杆菌，芽孢杆菌产生的粘多菌素。动植物产生的，蒜中的蒜素，动物脏器中的鱼素（ekmolin).根据作用谱分:广谱抗生素，如氨苄青霉素，可抑制G-&G+细菌抗G+菌的抗生素：如青霉素。抗G-菌的抗生素：如链霉素抗真菌的抗生素：如制霉菌素抗病毒的抗生素：如四环类抗生素对立克次体积较大病毒有一定的作用。抗癌抗生素：如阿霉素（Adriamycin)目前在抗病毒、抗癌、等方面还没有理想的抗生素。根据抗生素的结构分类以-内酰胺类抗生素：包括其青霉素、头孢菌素类等。它们含有一个四元内酰胺环，为目前最受重视的一类。,PenicillinStructure,B-内酰胺是抗生素具有抗菌活性所必需的，而R基团则赋予抗生素的稳定性、抗菌谱等的变化。,氨基糖苷类抗生素：包括链霉素、庆大霉素含有氨基糖苷类或氨基环醇四环类抗生素：四环素类、土霉素以四个苯为母核。多肽类抗生素：多粘菌素、杆菌肽由产孢子杆菌产生。蒽环类抗生素：阿霉素、柔红霉素属于抗癌类抗生素。喹诺酮类抗生素，如环丙沙星、诺氟沙星。按照作用机制分类抑制细胞壁合成的抗生素影响细胞膜功能的抗生素抑制病原菌蛋白质合成的抗生素，如四环素抑制核酸合成的抗生素如丝裂霉素C抑制生物功能作用的抗生素，,根据抗生素的生物合成途径分类氨基酸、肽类衍生物，如青霉素、头孢霉素等寡肽抗生素糖类衍生物，如链霉素等糖苷类抗生素以乙酸、丙酸为单位的衍生物，如红霉素等丙酸衍。生物,抗生素的应用,对抗生素的评价：有较大的差异毒力在体内发挥其抗生效能，而不被血液、脑肾液等破坏，不大量与血清蛋白质产生不可逆结合。给药后吸收快，并在被感染器官或组织中分布致病菌对抗生素不易产生耐药性不引起过敏反应理化性质稳定，便于提取、制剂和贮藏。抗生素的计量单位：效价单位：青霉素效价单位为能在50ml肉汤培养基中完全抑制金黄色葡萄球菌菌株发育的最小计量。,一个链霉菌效价单位为能在1ml肉汤培养基中完全抑制大肠杆菌标准菌株所需的最小链霉菌素计量。除了用效价单位之外，当物质被制成纯净的化学物质时，就可以用质量表示，而且与效价单位之间有一定的换算关系如1mg青霉素钠盐相当于1667个单位1mg链霉菌素碱相当于1000个单位,Industrialevolutionofpenicillinproduction,Commonantibioticsandtheirsources,BacitracinCephalosporin(s)（头孢菌素）Chloramphenicol（氯霉素）Cycloheximide（放线菌酮）HygromycinPenicillinStreptomycinTetracycline(s)Vancomycin,BacillussubtilisCephalosporiumsp.S.venezuelaeS.griseusS.hygromycesP.chrysogenumS.griseusS.aurofaciensS.orientalis,Beta-LactamStructure,半合成抗生素,用化学或生物化学的方法改变一直抗生素的化学结构或引入特定的功能基团而获得的新抗生素品种或其衍生物的总称。其目的：减低毒副作用，增强抗菌力，扩大抗菌谱，对耐药菌有效，便于口服，改善药理性质，提高生物有效性。,STRUTUREOFPENICILLIN,Whatisasemi-syntheticvariantofpenicillin?,DiscoveredthatnaturalpenicillinscanbechemicallymodifiedbyremovingtheRgroupandaddinganewonethatconfersnewpropertiesItisthevariationintheRgroupthatleadstothesemi-syntheticformsofpenicillin.Over20,000semi-syntheticvariants3mainvariantsofindustrialimportance,MainVariants,Ampicillin（氨卡青霉素）MethicillinAmoxycillin,Methicillin(甲氧霉素)1986（13%，1998，26%）,OriginallyintroducedtocombatS.AureusB-Lactamaseresistant1960sasemi-syntheticbroadspectrumcompoundproducedOverproductionofbeta-lactamaseenzymehasbeenshowntocontributetomethicillinresistance.,Ampicillin（氨卡青霉素）,Discoveredin1963OneofonlyfewpenicillinsthatcanbeadministeredorallyHasimprovedacidstabilityBroadspectrum.,Amoxycillin（羟氨卡青霉素）,Theprefferedchoiceofdrugfororaluse.ThehighestabsorbancelevelsinthebodyFirstmarketedin1998bySmitklineBeechams,Benefitsofsemi-syntheticvariants,CanbeadministeredorallyastheyareresistanttostomachacidTheyhaveabroaderspectrumofactivityThesemi-syntheticvariantofpenicillinischemicallymorestablethanthevolatilityoftheB-Lactamring,inthepenicillinG,6-APA（6-氨基青霉烷酸）,Firstdiscoveredin1959FirstindustrialprocessofenzymeengineeringConversionofPenGto6-APAUsedinpenicillinrecoveryprocess,Actionof6-APA,PGA（青霉素酰化酶）isthenaturalsubstrateforPen.GPenicillinGishydrolysedinareversiblereactionto6-APAandphenylaceticacid.Vitalintermediatefortheproductionofsemi-syntheicpenicillins,AcylGroup,NatureofacylgroupconferscertainpropertiesonthepenicllinanalogueSelectivehydrolysisoccursbyhydrolysisbylactamase-producingculture,lactamase,ThesearethemostcommoncauseofbacterialresistancetobetalactamantibioticsMostimportantlactamasewasproducedbyS.aureus.Thiswasresponsibleforriseinresistancetopenicillininthe1950sBetaLactamringiscrucialforpenicillintoremainactive,Actionoflactamase,lactamasedestroyspenicillinbyhydrolyzingabondinthisring,SomeStatistics,10,985tonnesof6-APAwereproducedin1998Ofthe10,985,48%wasusedforampicilllinproduction,and27%foramoxicillin.,Resistance,LinkedtooveruseofaspecificagentortotheintroductiobofanewclassoflactamBythelate1970sfewerthan10%ofStaphlococcusinfectionscouldbecuredbypenicillin，到1999年，WHO报道95%的金黄色葡萄球菌抗青霉素，100%大肠杆菌抗链霉素,Conclusion,Semi-syntheticvariantsarenowthefavouredchoiceofantibiticforcombatingbacterialresistanceCanbeorallyadministeredTheyhaveabroadenedrangeofspecifityMorechemicallystablethantheirnativecounterpart.,抗生素的生产和提取工艺,菌种,孢子制备,发酵,种子制备,发酵液的预处理,提取和精制,成品包装,现代抗生素工业生产工艺,Configuration&Production,ReactorsmustbecorrectlydesignedifthefermentingorganismspotentialistobeachievedAnunderstandingoftheimpactofconfigurationonproductioncanhelphighlightsuitabledesigns,BuildingaBioreactor,Themostimportantdesignfeaturestoconsiderinclude:ConstructionMaterialCapacityImpellarDesignHeatExchangerSpargerAnalyticalInstrumentation,TheOptimumDesign,ConstructionMaterialTypicallystainlesssteelasitscheap,light,readilyavailable&easytosterilizeCapacityIndustrialdesignsholdbetween30-200m3offluid,musthoweverconsiderimpactoncost&environmentalcontrol,GeometryIncreasedheighttodiameterratioisfavoredasitaidsagitation&aerationofviscousantibioticbrothsImpellarRushtonsarestandard,butaxialdesignsalsopossible.Lowrotationalspeedsminimizeshear&increasedimpellardiametercompensatesforreducedagitation,Axial&RadialImpellars,DifferentImpellarDesigns,HeatExchangerHeatgeneratedbybiologicalreactions&agitationisremovedbycoolingjackets.Limpet,orhalfpipe,iscommonchoiceSpargerC-shapedpreferabletosinglepipe,astheyimproveO2transferintheviscousbroths,ColdH2OIn,WarmH2OOut,AnalyticalInstrumentationProbesformeasurementoftemp,pHglucose&DOconc.arefittedatvariouspointsaroundthevesselOptimumtempbetween24-280CAlkalinepHmustbereducedtoneutralityGlucosemustbemonitoredtopreventcataboliterepressionDOmustsustaincellsbutpromotefermentation,抗生素的提取,一般用溶媒体取法：利用抗生素在不同的ph条件下的化学状态（游离酸，碱或盐）存在时，在水及与水互不相容的溶媒中溶解度不同的特性，使抗生素从一液相转移到另一液相中（如有机溶剂），已达到浓酸和提纯的目的。,DownstreamProcessing,StepsinDownstreamProcessing:,DownstreamProcessing,RemovalofInsolubles,-Filtration,Isolationofproduct,-SolventExtraction,Purification,-Precipitation,Electrophoresis,Polishing,-crystallisation,drying,PenicillinGisinsolutionexocellularly,Filtration,Removecellsbyfiltration,Sterilecondition