《抗生素发酵生产技术》由会员分享，可在线阅读，更多相关《抗生素发酵生产技术（34页珍藏版）》请在金锄头文库上搜索。
1、抗 生 素 发 酵 生 产 技 术Antibiotics Fermentation Production11. 概述2. 生产前期研究3. 生产菌改良4. 生产工艺过程5. 发酵条件的控制6. 提取和精制1. Introduction2. Research in Lab3. Strain improvement4. Production processing5. Fermentation control6. Isolation & purification 抗 生 素 发 酵 生 产 技 术Antibiotics Fermentation Production2抗生素是生物在其生产活动过程中所
2、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 Introduction3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
3、in 1929. World War II, fermented in large-scale The first major antibiotic to be commercialized1. 概述 Introduction4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) ：
4、 丝裂霉素 分类 Classification1. 概述 Introduction5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 (penicillins) 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 (tetracyclines) ： 四环素、土霉素多 肽 类 (polymyxin) ： 多粘菌素 分类 Classification1. 概述Introduction7 HNONSCH3CH3HCOO-H-lactam
5、ring -内酰胺 环Rnatural penicillin R =CH2-CO-8-内酰胺 环9两性霉素Antibiotics are so complex they can only be synthesized in a living system10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
6、Research in Lab11 Agar diffusion assayagar plate琼脂培养基lawn of test bacteria测 试 菌 苔filter papers soakedwith 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 electro
7、phoresis(CE)Gas chromatography (GC)High performance liquid chromatography (HPLC)Mass spectroscopy (MS) AutomationSensitivityAccuracy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 selectionPhysical mutagen eg. Ultraviolet RadiationChemical mutagen eg. Nitrosoguanidine (NTG)Impro
8、ved strain can be selected empirically 3. 抗生素生产菌改良 Strain Improvement14基因工程Genetic Engineering 3. 抗生素生产菌改良 Strain ImprovementRecombinant DNA Technology （DNA重组技术）克隆15Strain Improvement Scale-upStrain出发菌株Agar plate平板技术Primaryshake flaskEvaluation摇瓶培养Secondary shake flask evaluationdifferent media二级摇瓶培
9、养各种培养基Lab fermentor evaluationProductionCondition小型发酵罐模拟生产条件Mastercell bankWorkingcell bankInoculumDevelopment菌种扩大培养Pilot plantEvaluation中试生产ProductionEvaluation生产60003003031Possiblerecycle1617Scale-up（比拟放大）models EL Gadden Jr 200 m3 Same productivity ? Limitations ?模 拟18摇瓶发酵 Shake-flask fermentatio
10、n10-50 ml in 125-500 ml flaskBatch onlyLimited control: temperatureSlow metabolizing carbohydrateAmbient pressureBuffers needed to control pHIn-process sampling difficultVolume decrease by evaporationSolid growth on side wallsAntifoam not neededAgitation limited: shaker speedNo control of dissolved
11、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 fermentation10-100,000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 vesselsBatch and feed possibleContinuous control: pH, temperatureAmmonium salts or a
12、mmonia neededPressure at two atmospheres possibleNo buffers necessary In-process sampling easyVolume increase by sugar feedVery uniform growthAntifoam often requiredWide variety of impellers and bafflesDissolved oxygen controlled by aeration, pressure20摇 瓶 发 酵 Shake-flask 装液量：10% (250ml)间歇培养温控受限制代谢碳
13、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 product4. 抗生素生产工艺过程Anti
14、biotics Production Processing22A scheme of the material flow in an antibiotic production 23A 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 FermentationCarbon source:Beet & cane molassesGlucose, lactose, starchEthanol, methanol GlycerolMaltose syrup, corn s
15、yrupCottonseed oil, peanut oil, soy oilLard 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 liquorSoybean meal & flour Peanut meal Rapeseed mealBeef blood, Casein hyd
16、rolysate Whey Fish mealWhole yeastYeast extractMeat & bone meal 发酵生产原料 Materials for Fermentation26 培 养 基Media for Cell Growth & Final productionComponents Range % 组 成Glucose/ sucrose 3.0-5.0 葡萄糖/ 蔗糖Starch 淀粉 3.0-5.0Corn steep liquor 3.0-5.0Calcium carbonate 0.5-1.0 碳酸钙Phosphate 磷酸盐 0.1-0.5Ammonium sulphate 0.1-1.0 硫酸胺Urea 尿素 0.1-0.5Oil 油 0.1-0.5Fermentation 发酵培养基 Components Range % 组 成Glucose 0.5-1.0Starch 0.5-5.0Corn steep liquor 5.0-8.0 玉米浸提液Soy flour 1.0-5.0Phosphate 0.1-1.0Ammonium sulphate
