第九章 发酵工程各论---抗生素生产

Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,*,*,韩 北 忠,中国农业大学 食品学院,Han,Bei,-,zhong,Prof.,PhD,College of Food Science&Nutritional Engineering,China Agricultural University,抗 生 素 发 酵 生 产 技 术,Antibiotics Fermentation Production,1,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,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,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,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,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,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,根据抗生素的作用机制分类,(,Modes of Action of Antibiotics),抑制细胞壁合成,(,Cell Wall Synthesis),：青霉素,抑制细胞膜功能,(,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,复制,翻译,转录,6,根据抗生素的化学结构分类,-,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,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,-,内酰胺 环,9,两性霉素,Antibiotics are,so complex,they can only be synthesized,in a living system,10,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,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,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,分 析 仪 器,Analytical instruments,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,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,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,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,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,16,17,Scale-up,（,比拟放大）,models,EL,Gadden,Jr,200 m,3,Same productivity?,Limitations?,模 拟,18,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,容量减少,瓶壁上结垢,无需消泡,搅拌,受限制,无法控制溶解氧,19,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,多种搅拌,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,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,Stirred tank,21,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,A scheme of the material flow in an antibiotic production,23,A scheme of the material flow in an antibiotic production,连消,发酵罐,缓冲罐,种子罐,24,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,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,Rape