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(dòng)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(xì)胞得有機(jī)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(jī)制分類(ModesofActionofAntibiotics)抑制細(xì)胞壁合成(CellWallSynthesis)

:青霉素抑制細(xì)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(yàn)提取、精制、鑒定毒性試驗(yàn)藥理和臨床試驗(yàn)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(diǎn)

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(xì)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(dòng)靈敏準(zhǔn)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(kuò)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(bǔ)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(bǔ)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(bǔ)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(bǔ)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