抗生素發(fā)酵生產(chǎn)工藝學(xué)抗生素就是生物在其生產(chǎn)活動過程中所產(chǎn)生,并能在低微濃度下有選擇性地抑制或殺滅其她微生物或腫瘤細胞得有機物。

Antibiotics

Anyorganicsubstancesproducedbyvariousorganismsthathavethepowerofarrestingthegrowthofothermicroorganismsorofdestroyingthem、1、概述Introduction歷史1929年,英國弗萊明發(fā)現(xiàn)青霉素二戰(zhàn)期間,大規(guī)模發(fā)酵生產(chǎn)世界上最早用于臨床得抗菌素History

PenicillinwasdiscoveredbyFlemingin1929、WorldWarII,fermentedinlarge-scale

Thefirstmajorantibiotictobemercialized1、概述Introduction根據(jù)抗生素得作用機制分類(ModesofActionofAntibiotics)抑制細胞壁合成(CellWallSynthesis)

:青霉素抑制細胞膜功能(CellMembraneDamage)

:多烯類抗生素抑制蛋白質(zhì)合成(ProteinSynthesis)

:四環(huán)素抑制核酸合成(NucleicAcidSynthesis)

:絲裂霉素

分類Classification1、概述Introduction復(fù)制翻譯轉(zhuǎn)錄根據(jù)抗生素得化學(xué)結(jié)構(gòu)分類β-內(nèi)酰胺類(β-lactam):青霉素類(penicillins)

頭孢菌素類(cephalosporins)氨基糖苷類(aminoglycosides):鏈霉素、慶大霉素大環(huán)內(nèi)酯類(macrolides):紅霉素、麥迪加霉素四環(huán)類(tetracyclines):四環(huán)素、土霉素多肽類(polymyxin):多粘菌素

分類Classification1、概述Introduction

HNONSCH3CH3HCOO-H

-lactamringβ-內(nèi)酰胺環(huán)RnaturalpenicillinR=CH2-CO-β-內(nèi)酰胺環(huán)兩性霉素Antibioticsare

soplextheycanonlybesynthesized

inalivingsystem產(chǎn)生菌得篩選抗菌性試驗提取、精制、鑒定毒性試驗藥理和臨床試驗ScreeningofastrainAntimicrobialtrialIsolation,purificationandidentificationToxicologicaltrial

Medicinal&clinicaltrial

2、生產(chǎn)前期研究ResearchinLab大家有疑問的，可以詢問和交流可以互相討論下，但要小聲點

Agardiffusionassayagarplate瓊脂培養(yǎng)基lawnoftestbacteria測試菌苔filterpaperssoakedwithtestcompounds含藥物濾紙zonesofinhibition(nogrowth)

抑菌圈分析儀器Analyticalinstruments現(xiàn)代分析儀器:核磁共振毛細管電泳氣相色譜高效液相色譜質(zhì)譜Modernanalyticalinstruments:Nuclearmagneticresonance(NMR)Capillaryelectrophoresis

(CE)Gaschromatography

(GC)Highperformanceliquidchromatography

(HPLC)Massspectroscopy

(MS)

Automation

Sensitivity

Accuracy

自動靈敏準確人工誘變法:紫外線、X-線、r-線照射亞硝基胍、亞硝酸、秋水仙素、氮芥等誘變劑處理ConventionalstrainselectionPhysicalmutageneg、UltravioletRadiationChemicalmutageneg、Nitrosoguanidine(NTG)Improvedstraincanbeselectedempirically3、抗生素生產(chǎn)菌改良StrainImprovement基因工程GeneticEngineering

3、抗生素生產(chǎn)菌改良StrainImprovementRebinantDNATechnology(DNA重組技術(shù))克隆StrainImprovementScale-upStrain出發(fā)菌株Agarplate平板技術(shù)PrimaryshakeflaskEvaluation搖瓶培養(yǎng)Secondaryshakeflaskevaluationdifferentmedia二級搖瓶培養(yǎng)各種培養(yǎng)基

Labfermentorevaluation

ProductionCondition小型發(fā)酵罐模擬生產(chǎn)條件MastercellbankWorkingcellbankInoculumDevelopment菌種擴大培養(yǎng)PilotplantEvaluation中試生產(chǎn)ProductionEvaluation生產(chǎn)60003003031PossiblerecycleScale-up(比擬放大)modelsELGaddenJr<10dm3GoodproductGoodproductivity>200m3Sameproductivity?Limitations?模擬搖瓶發(fā)酵Shake-flaskfermentation10-50mlin125-500mlflaskBatchonlyLimitedcontrol:temperatureSlowmetabolizingcarbohydrateAmbientpressureBuffersneededtocontrolpHIn-processsamplingdifficultVolumedecreasebyevaporationSolidgrowthonsidewallsAntifoamnotneededAgitationlimited:shakerspeedNocontrolofdissolvedoxygen裝液量:10%(~250ml)間歇培養(yǎng)溫控受限制代謝碳水化合物慢常壓緩沖液控制pH在線取樣難容量減少瓶壁上結(jié)垢無需消泡攪拌受限制無法控制溶解氧攪拌罐發(fā)酵Stirredtankfermentation10-100,000升容量間歇和補料培養(yǎng)連續(xù)控制:pH,溫度需要銨鹽或氨氣2大氣壓無需緩沖液在線取樣易補料,容量增加生長均勻一致需消泡多種攪拌控制溶解氧:通氣,加壓10-100,000litervesselsBatchandfeedpossibleContinuouscontrol:pH,temperatureAmmoniumsaltsorammonianeededPressureattwoatmospherespossibleNobuffersnecessaryIn-processsamplingeasyVolumeincreasebysugarfeedVeryuniformgrowthAntifoamoftenrequiredWidevarietyofimpellersandbafflesDissolvedoxygencontrolledbyaeration,pressure搖瓶發(fā)酵Shake-flask裝液量:10%(~250ml)間歇培養(yǎng)溫控受限制代謝碳水化合物慢常壓緩沖液控制pH在線取樣難容量減少瓶壁上結(jié)垢無需消泡攪拌受限制無法控制溶解氧10-100,000升容量間歇和補料培養(yǎng)連續(xù)控制:pH,溫度需要銨鹽或氨氣2大氣壓無需緩沖液在線取樣易補料,容量增加生長均勻一致需消泡多種攪拌控制溶解氧:通氣,加壓攪拌罐發(fā)酵Stirredtank菌種→種子制備

→發(fā)酵

→發(fā)酵液預(yù)處理→提取及精制→成品包裝Strain→Preparationofseed

→Fermentation

→Treatmentofbroth→Isolation&purification→Packageofproduct4、抗生素生產(chǎn)工藝過程AntibioticsProductionProcessingAschemeofthematerialflowinanantibioticproductionAschemeofthematerialflowinanantibioticproduction連消發(fā)酵罐緩沖罐種子罐發(fā)酵生產(chǎn)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,甲醇甘油麥芽糖漿,玉米糖漿綿籽油,花生油,豆油豬油油酸甲酯油脂氮源:綿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,酪蛋白水解液乳清魚粉酵母酵母膏肉粉和骨粉

Nitrogensource:CottonseedmealCorn

meal&steepliquorSoybeanmeal&flourPeanutmealRapeseedmealBeefblood,CaseinhydrolysateWheyFishmealWholeyeastYeastextractMeat&bonemeal

發(fā)酵生產(chǎn)原料

MaterialsforFermentation

培養(yǎng)基MediaforCellGrowth&FinalproductionponentsRange%組成Glucose/sucrose3、0-5、0葡萄糖/蔗糖Starch淀粉3、0-5、0Cornsteepliquor3、0-5、0Calciumcarbonate0、5-1、0碳酸鈣Phosphate磷酸鹽0、1-0、5Ammoniumsulphate0、1-1、0

硫酸胺Urea尿素0、1-0、5Oil油0、1-0、5Fermentation發(fā)酵培養(yǎng)基

ponentsRange%組成Glucose0、5-1、0Starch

0、5-5、0Cornsteepliquor5、0-8、0玉米浸提液Soyflour1、0-5、0Phosphate