资源描述
Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,*,*,韩 北 忠,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,生产菌改进,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，并能在卑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.抗生素生产菌改进,Strain Improvement,14,基因工程,Genetic Engineering,3.抗生素生产菌改进,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（比较放大）,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,Rapeseed meal,Beef blood,Casein,hydrolysate,Whe
展开阅读全文