抗生素发酵生产工艺学jsp

概述生产前期研究生产菌改良生产工艺过程发酵条件的控制提取和精制IntroductionResearchinLabStrainimprovementProductionprocessingFermentationcontrolIsolation&purification抗生素发酵生产技术AntibioticsFermentationProduction1当前1页，总共33页。抗生素是生物在其生产活动过程中所产生，并能在低微浓度下有选择性地抑制或杀灭其他微生物或肿瘤细胞的有机物。

Antibiotics

Anyorganicsubstancesproducedbyvariousorganismsthathavethepowerofarrestingthe

growthofothermicroorganismsorofdestroyingthem.1.概述Introduction2当前2页，总共33页。历史1929年，英国弗莱明发现青霉素二战期间，大规模发酵生产世界上最早用于临床的抗菌素History

PenicillinwasdiscoveredbyFlemingin1929.WorldWarII,fermentedinlarge-scale

Thefirstmajorantibiotictobecommercialized1.概述Introduction3当前3页，总共33页。根据抗生素的作用机制分类(ModesofActionofAntibiotics)

抑制细胞壁合成(CellWallSynthesis)

：青霉素抑制细胞膜功能(CellMembraneDamage)

：多烯类抗生素抑制蛋白质合成(ProteinSynthesis)

：四环素抑制核酸合成(NucleicAcidSynthesis)

：丝裂霉素

分类Classification1.概述Introduction4当前4页，总共33页。复制翻译转录5当前5页，总共33页。根据抗生素的化学结构分类β-内酰胺类(β-lactam)：青霉素类(penicillins)

头孢菌素类(cephalosporins)氨基糖苷类(aminoglycosides)：链霉素、庆大霉素大环内酯类(macrolides)：红霉素、麦迪加霉素四环类(tetracyclines)：四环素、土霉素多肽类(polymyxin)：多粘菌素

分类Classification1.概述Introduction6当前6页，总共33页。

HNONSCH3CH3HCOO-H-lactamringβ-内酰胺环RnaturalpenicillinR=CH2-CO-7当前7页，总共33页。β-内酰胺环8当前8页，总共33页。两性霉素Antibioticsare

socomplextheycanonlybesynthesized

inalivingsystem9当前9页，总共33页。产生菌的筛选抗菌性试验提取、精制、鉴定毒性试验药理和临床试验ScreeningofastrainAntimicrobialtrialIsolation,purificationandidentificationToxicologicaltrial

Medicinal&clinicaltrial

2．生产前期研究ResearchinLab10当前10页，总共33页。

Agardiffusionassayagarplate琼脂培养基lawnoftestbacteria测试菌苔filterpaperssoakedwithtestcompounds含药物滤纸zonesofinhibition(nogrowth)

抑菌圈11当前11页，总共33页。分析仪器Analyticalinstruments现代分析仪器：核磁共振毛细管电泳气相色谱高效液相色谱质谱Modernanalyticalinstruments:Nuclearmagneticresonance(NMR)Capillaryelectrophoresis

(CE)Gaschromatography

(GC)Highperformanceliquidchromatography

(HPLC)Massspectroscopy

(MS)

Automation

Sensitivity

Accuracy

自动灵敏准确12当前12页，总共33页。人工诱变法：紫外线、X-线、r-线照射亚硝基胍、亚硝酸、秋水仙素、氮芥等诱变剂处理ConventionalstrainselectionPhysicalmutageneg.UltravioletRadiationChemicalmutageneg.Nitrosoguanidine(NTG)Improvedstraincanbeselectedempirically3.抗生素生产菌改良StrainImprovement13当前13页，总共33页。基因工程GeneticEngineering

3.抗生素生产菌改良StrainImprovementRecombinantDNATechnology（DNA重组技术）克隆14当前14页，总共33页。StrainImprovementScale-upStrain出发菌株Agarplate平板技术PrimaryshakeflaskEvaluation摇瓶培养Secondaryshakeflaskevaluationdifferentmedia二级摇瓶培养各种培养基

Labfermentorevaluation

ProductionCondition小型发酵罐模拟生产条件MastercellbankWorkingcellbankInoculumDevelopment菌种扩大培养PilotplantEvaluation中试生产ProductionEvaluation生产60003003031Possiblerecycle15当前15页，总共33页。16当前16页，总共33页。Scale-up（比拟放大）modelsELGaddenJr<10dm3GoodproductGoodproductivity>200m3Sameproductivity?Limitations?模拟17当前17页，总共33页。摇瓶发酵Shake-flaskfermentation10-50mlin125-500mlflaskBatchonlyLimitedcontrol:temperatureSlowmetabolizingcarbohydrateAmbientpressureBuffersneededtocontrolpHIn-processsamplingdifficultVolumedecreasebyevaporationSolidgrowthonsidewallsAntifoamnotneededAgitationlimited:shakerspeedNocontrolofdissolvedoxygen装液量：10%(~250ml)间歇培养温控受限制代谢碳水化合物慢常压缓冲液控制pH在线取样难容量减少瓶壁上结垢无需消泡搅拌受限制无法控制溶解氧18当前18页，总共33页。搅拌罐发酵Stirredtankfermentation10-100,000升容量间歇和补料培养连续控制：pH，温度需要铵盐或氨气2大气压无需缓冲液在线取样易补料,容量增加生长均匀一致需消泡多种搅拌控制溶解氧:通气,加压10-100,000litervesselsBatchandfeedpossibleContinuouscontrol:pH,temperatureAmmoniumsaltsorammonianeededPressureattwoatmospherespossibleNobuffersnecessaryIn-processsamplingeasyVolumeincreasebysugarfeedVeryuniformgrowthAntifoamoftenrequiredWidevarietyofimpellersandbafflesDissolvedoxygencontrolledbyaeration,pressure19当前19页，总共33页。摇瓶发酵Shake-flask装液量：10%(~250ml)间歇培养温控受限制代谢碳水化合物慢常压缓冲液控制pH在线取样难容量减少瓶壁上结垢无需消泡搅拌受限制无法控制溶解氧10-100,000升容量间歇和补料培养连续控制：pH，温度需要铵盐或氨气2大气压无需缓冲液在线取样易补料,容量增加生长均匀一致需消泡多种搅拌控制溶解氧:通气,加压搅拌罐发酵Stirredtank20当前20页，总共33页。菌种

→种子制备

→发酵

→发酵液预处理→提取及精制→成品包装Strain

→Preparationofseed

→Fermentation

→Treatmentofbroth→Isolation&purification→Packageofproduct4.抗生素生产工艺过程AntibioticsProductionProcessing21当前21页，总共33页。Aschemeofthematerialflowinanantibioticproduction22当前22页，总共33页。Aschemeofthematerialflowinanantibioticproduction连消发酵罐缓冲罐种子罐23当前23页，总共33页。发酵生产原料

MaterialsforFermentationCarbonsource:Beet&canemolassesGlucose,lactose,starchEthanol,methanolGlycerolMaltosesyrup,cornsyrupCottonseedoil,peanutoil,soyoilLardoilMethyloleateTallow碳源:甜菜和甘蔗糖蜜葡萄糖，乳糖，淀粉乙醇，甲醇甘油麦芽糖浆，玉米糖浆绵籽油，花生油，豆油猪油油酸甲酯油脂24当前24页，总共33页。氮源:绵籽粉玉米粉和玉米浸出液大豆粉和大豆面花生粉菜籽粉牛血，酪蛋白水解液乳清鱼粉酵母酵母膏肉粉和骨粉

Nitrogensource:CottonseedmealCorn

meal&steepliquorSoybeanmeal&flourPeanutmealRapeseedmealBeefblood,CaseinhydrolysateWheyFishmealWholeyeastYeastextractMeat&bonemeal

发酵生产原料

MaterialsforFermentation25当前25页，总共33页。

培养基MediaforCellGrowth&FinalproductionComponentsRange%组成葡萄糖/蔗糖Starch淀粉碳酸钙Phosphate磷酸盐

硫酸胺Urea尿素Oil油Fermentation发酵培养基

ComponentsRange%组成Starch

玉米浸提液andfedperiodicallyOil1.0andfedcontinuouslyCornsyrupfedcontinuously

玉米糖浆连续加料Tankseed种子培养基26当前26页，总共33页。StirredtankFermentation搅拌发酵27当前27页，总共33页。5.发酵控制FermentationControl

SampleAnalysispHDOSugarAmmoniaPhosphateSulphateProductsPrecursorsContaminationPressureprobeLevelprobepHprobeTbeDOprobeAntifoamAcid/BaseCoolingAir/agitationSugar/Oilfeed28当前28页，总共33页。ScaleWeightSignalPumpSpeedAmpli