抗生素发酵生产技术.pptx

Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,*,Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,*,*,Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,*,*,抗 生 素 发 酵 生 产 技 术,Antibiotics Fermentation Production,1,抗生素发酵生产技术,第1页,概述,生产前期研究,生产菌改良,生产工艺过程,发酵条件控制,提取和精制,Introduction,Research in Lab,Strain improvement,Production processing,Fermentation control,Isolation&purification,抗 生 素 发 酵 生 产 技 术,Antibiotics Fermentation Production,2,抗生素发酵生产技术,第2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,其它微生物或肿瘤细胞,有机物,。,Antibiotics,Any,organic substances,produced by various organisms that have the power of,arresting,the,growth of,other microorganisms,or of,destroying,them.,1.,概述,Introduction,3,抗生素发酵生产技术,第3页,历史,1929,年，英国弗莱明发觉青霉素,二战期间，大规模发酵生产,世界上最早用于临床抗菌素,History,Penicillin was discovered by Fleming in 1929.,World War II,fermented in large-scale,The first major antibiotic to be commercialized,1.,概述,Introduction,4,抗生素发酵生产技术,第4页,依据抗生素作用机制分类,(Modes of Action of Antibiotics),抑制细胞壁合成,(Cell Wall Synthesis),：青霉素,抑制细胞膜功效,(Cell Membrane Damage),：多烯类抗生素,抑制蛋白质合成,(Protein Synthesis),：四环素,抑制核酸合成,(Nucleic Acid Synthesis),：丝裂霉素,分类,Classification,1.,概述,Introduction,5,抗生素发酵生产技术,第5页,复制,翻译,转录,6,抗生素发酵生产技术,第6页,依据抗生素化学结构分类,-,内酰胺类,(-lactam),：青霉素类,(p,enicillins),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,(t,etracyclines),：四环素、土霉素,多 肽 类,(polymyxin),：多粘菌素,分类,Classification,1.,概述,Introduction,7,抗生素发酵生产技术,第7页,H,N,O,N,S,CH,3,CH,3,H,COO-,H,-,lactam ring,-,内酰胺 环,R,natural penicillin,R=,CH,2,-CO-,8,抗生素发酵生产技术,第8页,-,内酰胺 环,9,抗生素发酵生产技术,第9页,两性霉素,Antibiotics are,so complex,they can only be synthesized,in a living system,10,抗生素发酵生产技术,第10页,产生菌筛选,抗菌性试验,提取、精制、判定,毒性试验,药理和临床试验,Screening of a strain,Antimicrobial trial,Isolation,purification and identification,Toxicological trial,Medicinal&clinical trial,2,生产前期研究,Research in Lab,11,抗生素发酵生产技术,第11页,Agar diffusion assay,agar plate,琼脂培养基,lawn of test bacteria,测 试 菌 苔,filter papers soaked,with test compounds,含 药 物 滤 纸,zones of inhibition,(,no growth),抑菌圈,12,抗生素发酵生产技术,第12页,分 析 仪 器,Analytical instruments,当代分析仪器：,核磁共振,毛细管电泳,气相色谱,高效液相色谱,质谱,Modern analytical instruments:,Nuclear magnetic resonance(,NMR,),Capillary electrophoresis,(,CE,),Gas chromatography,(,GC,),High performance liquid chromatography,(,HPLC,),Mass spectroscopy,(,MS,),Automation,Sensitivity,Accuracy,自动灵敏准确,13,抗生素发酵生产技术,第13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,Conventional strain selection,Physical mutagen,eg.,Ultraviolet Radiation,Chemical mutagen,eg.,Nitrosoguanidine,(NTG),Improved strain can be selected empirically,3.,抗生素,生产菌改良,Strain Improvement,14,抗生素发酵生产技术,第14页,基因工程,Genetic Engineering,3.,抗生素,生产菌改良,Strain Improvement,Recombinant DNA Technology,（,DNA,重组技术,）,克隆,15,抗生素发酵生产技术,第15页,Strain Improvement Scale-up,Strain,出发菌株,Agar plate,平板技术,Primary,shake flask,Evaluation,摇瓶培养,Secondary,shake flask,evaluation,different media,二级摇瓶培养,各种培养基,Lab fermentor,evaluation,Production,Condition,小型发酵罐,模拟生产条件,Master,cell bank,Working,cell bank,Inoculum,Development,菌种扩大培养,Pilot plant,Evaluation,中试生产,Production,Evaluation,生产,6000,300,30,3,1,Possible,recycle,16,抗生素发酵生产技术,第16页,17,抗生素发酵生产技术,第17页,Scale-up,（,比拟放大）,models,EL Gadden Jr,200 m,3,Same productivity?,Limitations?,模 拟,18,抗生素发酵生产技术,第18页,摇瓶发酵,Shake-flask fermentation,10-50 ml in 125-500 ml flask,Batch only,Limited control:temperature,Slow metabolizing carbohydrate,Ambient pressure,Buffers needed to control pH,In-process sampling difficult,Volume decrease by evaporation,Solid growth on side walls,Antifoam not needed,Agitation limited:shaker speed,No control of dissolved oxygen,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,容量降低,瓶壁上结垢,无需消泡,搅拌受限制,无法控制溶解氧,19,抗生素发酵生产技术,第19页,搅 拌 罐 发 酵,Stirred tank fermentation,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,各种搅拌,控制溶解氧,:,通气,加压,10-100,000 liter vessels,Batch and feed possible,Continuous control:pH,temperature,Ammonium salts or ammonia needed,Pressure at two atmospheres possible,No buffers necessary,In-process sampling easy,Volume increase by sugar feed,Very uniform growth,Antifoam often required,Wide variety of impellers and baffles,Dissolved oxygen controlled by,aeration,pressure,20,抗生素发酵生产技术,第20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,容量降低,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,各种搅拌,控制溶解氧,:,通气,加压,搅 拌 罐 发 酵,Stirred tank,21,抗生素发酵生产技术,第21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,Preparation of seed,Fermentation,Treatment of broth,Isolation&purification,Package of product,4.,抗生素生产工艺过程,Antibiotics Production Processing,22,抗生素发酵生产技术,第22页,A scheme of the material flow in an antibiotic production,23,抗生素发酵生产技术,第23页,A scheme of the material flow in an antibiotic production,连消,发酵罐,缓冲罐,种子罐,24,抗生素发酵生产技术,第24页,发酵生产原料,Materials for Fermentation,Carbon source:,Beet&cane molasses,Glucose,lactose,starch,Ethanol,methanol,Glycerol,Maltose syrup,corn syrup,Cottonseed oil,peanut oil,soy oil,Lard oil,Methyl oleate,Tallow,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,25,抗生素发酵生产技术,第25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,Nitrogen source:,Cotton seed meal,Corn,meal&steep liquor,Soybean meal&flour,Peanut meal,Rapeseed meal,Beef blood,Casein hydrolysate,Whey,Fish meal,Whole yeast,Yeast extract,Meat&bone meal,发酵生产原料,Materials for Fermentation,26,抗生素发酵生产技术,第26页,培 养 基,Media for Cell Growth&Final production,Components Range%,组 成,Glucose/sucrose 3.0-5.0,葡萄糖,/,蔗糖,Starch,淀粉,3.0-5.0,Corn steep liquor 3.0-5.0,Calcium carbonate 0.5-1.0,碳酸钙,Phosphate,磷酸盐,0.1-0.5,Ammonium sulphate 0.1-1.0,硫酸胺,Urea,尿素,0.1-0.5,Oil,油,0.1-0.5,Fermentation,发酵培养基,Components Range%,组 成,Glucose 0.5-1.0,Starch,0.5-5.0,Corn steep liquor 5.0-8.0,玉米浸提液,Soy flour 1.0-5.0,Phosphate 0.1-1.0,Ammonium sulphate 0.5-1.0,and fed periodically,Oil 1.0,and fed continuously,Corn syrup fed continuously,玉米糖浆 连续加料,Tank seed,种子培养基,27,抗生素发酵生产技术,第27页,Stirred tank Fermentation,搅拌发酵,28,抗生素发酵生产技术,第28页,5.,发 酵 控 制,Fermentation Control,Sample Analysis,pH,DO,Sugar,Ammonia,Phosphate,Sulphate,Products,Precursors,Contamination,Pressure probe,Level probe,pH probe,Temp.probe,DO probe,Antifoam,Acid/Base,Cooling,Air/agitation,Sugar/Oil,feed,29,抗生素发酵生产技术,第29页,Scale,Weight Signal,Pump Speed,Amplifier,Controller,Supervisory,Computer,Flow Rate Control,流量控制,30,抗生素发酵生产技术,第30页,发 酵 控 制,Fermentation control,31,抗生素发酵生产技术,第31页,菌种,(strain),：产黄青霉菌,(,Penicillium chrysogenum,),孢子培养,(spore cultivation),：,25,C,6-7 d,种子培养,(seed cultivation),：,25,C,，,2-3 d,发酵培养,(fermentation),：,22-26,C,6-7 d,加糖控制,(sugar feeding),：残糖降至,0.6,青 霉 素 发 酵,(1),Penicillin Fermentation,32,抗生素发酵生产技术,第32页,补氮,(nitrogen feeding),：氨控制在,0.01-0.05,前体,(precursor),：残余苯乙酰胺,0.05-0.08,温度控制,(temperature),：前期,25-26,C,后期,23,C,通气与搅拌,(aeration&stir),：,1:0.8 VVM,泡沫,(foam),：天然油脂,如豆油,青 霉 素 发 酵,(2),Penicillin Fermentation,33,抗生素发酵生产技术,第33页,Penicillin Fermentation Profile,改变曲线,34,抗生素发酵生产技术,第34页,