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,β-,内酰胺类,(β-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,β-,内酰胺 环,R,natural penicillin,R =,CH,2,-CO-,8,β-,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,的筛选,抗菌性,试,试验,提取、,精,精制、,鉴,鉴定,毒性试,验,验,药理和,临,临床试,验,验,Screeningofastrain,Antimicrobial trial,Isolation, purificationandidentification,Toxicological trial,Medicinal&clinicaltrial,2,．,生产前,期,期研究,ResearchinLab,11,Agar diffusionassay,agar plate,琼脂培,养,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,仪,器,器,Analytical instruments,现代分,析,析仪器,：,：,核磁共,振,振,毛细管,电,电泳,气相色,谱,谱,高效液,相,相色谱,质谱,Modernanalyticalinstruments:,Nuclear magnetic resonance(NMR),Capillaryelectrophoresis(CE),Gaschromatography(GC),High performance liquidchromatography(HPLC),Mass spectroscopy(MS),,AutomationSensitivityAccuracy,自 动,灵,灵 敏,准,准 确,13,人工诱,变,变法：,紫外线、,X-,线、,r-,线照射,亚硝基,胍,胍、亚硝,酸,酸、秋,水,水仙素,、,、氮芥,等,等诱变,剂,剂处理,Conventionalstrain selection,Physicalmutagen,eg.UltravioletRadiation,Chemicalmutagen,eg.Nitrosoguanidine(NTG),Improvedstraincan be selected empirically,3.,抗生素,生产菌,改,改良,StrainImprovement,14,基因工,程,程,Genetic Engineering,,,3.,抗生素,生产菌,改,改良,StrainImprovement,RecombinantDNATechnology,（,DNA,重组技,术,术,）,克隆,15,StrainImprovementScale-up,Strain,出发菌,株,株,Agar plate,平板技,术,术,Primary,shakeflask,Evaluation,摇瓶培,养,养,Secondary,shakeflask,evaluation,differentmedia,二级摇,瓶,瓶培养,各种培,养,养基,,Labfermentor,evaluation,Production,Condition,小型发,酵,酵罐,模拟生,产,产条件,Master,cell bank,Working,cell bank,Inoculum,Development,菌种扩,大,大培养,Pilotplant,Evaluation,中试生,产,产,Production,Evaluation,生产,6000,300,30,3,1,Possible,recycle,16,17,Scale-up,（,比拟放,大,大）,models,EL Gadden Jr,<,10,dm,3,Good product,Good productivity,> 200 m,3,Same productivity ?,Limitations ?,模,拟,拟,18,摇 瓶,发,发,酵,酵,Shake-flaskfermentation,10-50mlin125-500 ml flask,Batchonly,Limited control: temperature,Slow metabolizingcarbohydrate,Ambient pressure,Buffers neededtocontrol pH,In-process sampling difficult,Volumedecreasebyevaporation,Solidgrowthonsidewalls,Antifoamnotneeded,Agitationlimited:shaker speed,Nocontrolofdissolvedoxygen,装,液,液,量,量,：,：,10%(~250ml),间,歇,歇,培,培,养,养,温,控,控,受,受,限,限,制,制,代,谢,谢,碳,碳,水,水,化,化,合,合,物,物,慢,慢,常,压,压,缓,冲,冲,液,液,控,控,制,制,pH,在,线,线,取,取,样,样,难,难,容,量,量,减,减,少,少,瓶,壁,壁,上,上,结,结,垢,垢,无,需,需,消,消,泡,泡,搅,拌,拌,受,受,限,限,制,制,无,法,法,控,控,制,制,溶,溶,解,解,氧,氧,19,搅,拌,拌,罐,罐,发,发,酵,酵,Stirredtankfermentation,10-100,000,升,容,容,量,量,间,歇,歇,和,和,补,补,料,料,培,培,养,养,连,续,续,控,控,制,制,：,：,pH,，,温,度,度,需,要,要,铵,铵,盐,盐,或,或,氨,氨,气,气,2,大,气,气,压,压,无,需,需,缓,冲,冲,液,液,在,线,线,取,取,样,样,易,易,补,料,料,,,容,量,量,增,增,加,加,生,长,长,均,均,匀,匀,一,一,致,致,需,消,消,泡,泡,多,种,种,搅,搅,拌,拌,控,制,制,溶,溶,解,解,氧,氧,:,通,气,气,,,加,压,压,,10-100,000litervessels,Batchandfeedpossible,Continuouscontrol:pH,temperature,Ammoniumsaltsorammonianeeded,Pressureattwoatmospherespossible,Nobuffersnecessary,In-processsamplingeasy,Volumeincreasebysugarfeed,Veryuniformgrowth,Antifoamoftenrequired,Widevarietyofimpellersandbaffles,Dissolvedoxygencontrolledby,aeration,pressure,20,摇,瓶,瓶,发,发,酵,酵,Shake-flask,装,液,液,量,量,：,：,10%(~250ml),间,歇,歇,培,培,养,养,温,控,控,受,受,限,限,制,制,代,谢,谢,碳,碳,水,水,化,化,合,合,物,物,慢,慢,常,压,压,缓,冲,冲,液,液,控,控,制,制,pH,在,线,线,取,取,样,样,难,难,容,量,量,减,减,少,少,瓶,壁,壁,上,上,结,结,垢,垢,无,需,需,消,消,泡,泡,搅,拌,拌,受,受,限,限,制,制,无,法,法,控,控,制,制,溶,溶,解,解,氧,氧,10-100,000,升,容,容,量,量,间,歇,歇,和,和,补,补,料,料,培,培,养,养,连,续,续,控,控,制,制,：,：,pH,，,温,度,度,需,要,要,铵,铵,盐,盐,或,或,氨,氨,气,气,2,大,气,气,压,压,无,需,需,缓,冲,冲,液,液,在,线,线,取,取,样,样,易,易,补,料,料,,,容,量,量,增,增,加,加,生,长,长,均,均,匀,匀,一,一,致,致,需,消,消,泡,泡,多,种,种,搅,搅,拌,拌,控,制,制,溶,溶,解,解,氧,氧,:,通,气,气,,,加,压,压,搅,拌,拌,罐,罐,发,发,酵,酵,Stirredtank,21,菌,种,种,→,种,子,子,制,制,备,备,→,发,酵,酵,→,发,酵,酵,液,液,预,预,处,处,理,理,→,提,取,取,及,及,精,精,制,制,→,成,品,品,包,包,装,装,Strain,→,Preparationofseed,→,Fermentation,→,Treatmentofbroth,→,Isolation&purification,→,Packageofproduct,4.,抗,生,生,素,素,生,生,产,产,工,工,艺,艺,过,过,程,程,AntibioticsProductionProcessing,22,A schemeofthematerialflow inanantibiotic production,23,A schemeofthematerialflow inanantibiotic production,连消,发酵罐,缓冲罐,种子罐,24,发酵生产,原,原料,Materials for Fermentation,Carbon source:,Beet& canemolasses,Glucose,lactose, starch,Ethanol,methanol,Glycerol,Maltosesyrup, cornsyrup,,Cottonseed oil,peanut oil,soyoil,Lardoil,Methyl oleate,Tallow,碳源,:,甜菜和甘,蔗,蔗糖蜜,葡萄糖，,乳,乳糖，淀,粉,粉,乙醇，甲,醇,醇,甘油,麦芽糖浆,，,，玉米糖,浆,浆,,绵,籽,油，花生,油,油，豆油,猪油,油酸甲酯,油脂,25,氮源,:,绵,籽,粉,玉米粉和,玉,玉米浸出,液,液,大豆粉和,大,大豆面,花生粉,菜籽粉,,牛血，酪,蛋,蛋白水解,液,液,乳清,鱼粉,酵母,酵母膏,肉粉和骨,粉,粉,Nitrogensource:,Cottonseedmeal,Cornmeal&steepliquor,Soybeanmeal&flour,Peanutmeal,Rapeseedmeal,,Beefblood,Caseinhydrolysate,Whey,Fishmeal,Wholeyeast,Yeastextract,Meat&bonemeal,发,酵,酵,生,生,产,产,原,原,料,料,MaterialsforFermentation,26,培,养,养,基,基,MediaforCellGrowth&Finalproduction,ComponentsRange%,组,成,成,Glucose/sucrose3.0-5.0,葡,萄,萄,糖,糖,/,蔗,糖,糖,Starch,淀,粉,粉,3.0-5.0,Cornsteepliquor3.0-5.0,Calciumcarbonate0.5-1.0,碳,酸,酸,钙,钙,Phosphate,磷,酸,酸,盐,盐,0.1-0.5,Ammoniumsulphate0.1-1.0,硫,酸,酸,胺,胺,Urea,尿,素,素,0.1-0.5,Oil,油,0.1-0.5,,Fermentation,发,酵,酵,培,培,养,养,基,基,ComponentsRange%,组,成,成,Glucose0.5-1.0,Starch0.5-5.0,Cornsteepliquor5.0-8.0,玉,米,米,浸,浸,提,提,液,液,Soyflour1.0-5.0,Phosphate0.1-1.0,Ammoniumsulphate0.5-1.0,andfedperiodically,Oil1.0,andfedcontinuously,Cornsyrupfedcontinuously,玉,米,米,糖,糖,浆,浆,连,连,续,续,加,加,料,料,,Tankseed,种,子,子,培,培,养,养,基,基,27,StirredtankFermentation,搅,拌,拌,发,发,酵,酵,28,5.,发,酵,酵,控,控,制,制,FermentationControl,SampleAnalysis,pH,DO,Sugar,Ammonia,Phosphate,Sulphate,Products,Precursors,Contamination,,Pressureprobe,Levelprobe,pHprobe,Temp.probe,DOprobe,Antifoam,Acid/Base,Cooling,Air/agitation,Sugar/Oil,feed,29,,Scale,WeightSignal,PumpSpeed,Amplifier,Controller,Supervisory,Computer,FlowRateControl,流,量,量,控,控,制,制,30,发,酵,酵,控,控,制,制,Fermentationcontrol,31,菌,种,种,(strain),：,产,产,黄,黄,青,青,霉,霉,菌,菌,(,Penicilliumchrysogenum,),孢,子,子,培,培,养,养,(sporecultivation),：,25,ºC,,6-7d,种,子,子,培,培,养,养,(seedcultivation),：,25,ºC,，,2-3d,发,酵,酵,培,培,养,养,(fermentation),：,22-26,ºC,,6-7d,加,糖,糖,控,控,制,制,(sugarfeeding),：,残,残,糖,糖,降,降,至,至,0.6,％,,青,霉,霉,素,素,发,发,酵,酵,(1),PenicillinFermentation,32,补,氮,氮,(nitrogenfeeding),：,氨,氨,控,控,制,制,在,在,0.01-0.05,％,前,体,体,(precursor),：,残,残,余,余,苯,苯,乙,乙,酰,酰,胺,胺,0.05-0.08,％,温,度,度,控,控,制,制,(temperature),：,前,前,期,期,25-26,ºC,后,期,期,23,ºC,通,气,气,与,与,搅,搅,拌,拌,(aeration&stir),：,1:0.8VVM,泡,沫,沫,(foam),：,天,天,然,然,油,油,脂,脂,,,如,豆,豆,油,油,,青,霉,霉,素,素,发,发,酵,酵,(2),PenicillinFermentation,33,PenicillinFermentationProfile,变,化,化,曲,曲,线,线,34,9,、,静,静,夜,夜,四,四,无,无,邻,邻,，,，,荒,荒,居,居,旧,旧,业,业,贫,贫,。
12,月,月-2212,月,月-22,Thursday,December29,2022,10,、,雨,雨,中,中,黄,黄,叶,叶,树,树,，,，,灯,灯,下,下,白,白,头,头,人,人,04:48:2004:48:2004:48,12/29/20224:48:20AM,11,、以,我,我独,沈,沈久,，,，愧,君,君相,见,见频,12,月,月-2204:48:2004:48,Dec-2229-Dec-22,12,、故,人,人江,海,海别,，,，几,度,度隔,山,山川,04:48:2004:48:2004:48,Thursday,December29,2022,13,、乍见翻,疑,疑梦，相,悲,悲各问年,12月-2212月-2204:48:2004:48:20,December29,2022,14,、他乡生,白,白发，旧,国,国见青山,29 十,二,二月 20224:48:20,上,上午04:48:2012月-22,15,、比不,了,了得就,不,不比，,得,得不到,的,的就不,要,要十二月224:48 上,午,午12月-2204:48,December29,2022,16,、行动,出,出成果,，,，工作,出,出财富,。
2022/12/29 4:48:2004:48:20,29December2022,17,、做,前,前，,能,能够,环,环视,四,四周,；,；做,时,时，,你,你只,能,能或,者,者最,好,好沿,着,着以,脚,脚为,起,起点,的,的射,线,线向,前,前4:48:20,上,上午4:48,上,上,午,午04:48:2012,月,月-22,9,、没有,失,失败，,只,只有暂,时,时停止,成,成功！,12月-2212月-22,Thursday,December29, 2022,10,、很多事,情,情努力了,未,未必有结,果,果，但是,不,不努力却,什,什么改变,也,也没有04:48:2004:48:2004:48,12/29/2022 4:48:20 AM,11,、成,功,功就,是,是日,复,复一,日,日那,一,一点,点,点小,小,小努,力,力的,积,积累,12,月,月-2204:48:2004:48,Dec-2229-Dec-22,12,、,世,世,间,间,成,成,事,事,，,，,不,不,求,求,其,其,绝,绝,对,对,圆,圆,满,满,，,，,留,留,一,一,份,份,不,不,足,足,，,，,可,可,得,得,无,无,限,限,完,完,美,美,。
04:48:2004:48:2004:48,Thursday,December29,2022,13,、不,知,知香,积,积寺,，,，数,里,里入,云,云峰,12,月,月-2212,月,月-2204:48:2004:48:20,December29,2022,14,、意志坚,强,强的人能,把,把世界放,在,在手中像,泥,泥块一样,任,任意揉捏,29 十,二,二月 20224:48:20,上,上午04:48:2012月-22,15,、楚塞,三,三湘接,，,，荆门,九,九派通,十二月224:48 上,午,午12月-2204:48,December29,2022,16,、少年十,五,五二十时,，,，步行夺,得,得胡马骑,2022/12/29 4:48:2004:48:20,29 December2022,17,、空山新,雨,雨后，天,气,气晚来秋,4:48:20,上,上午4:48,上,上午04:48:2012月-22,9,、,杨,杨,柳,柳,散,散,和,和,风,风,，,，,青,青,山,山,澹,澹,吾,吾,虑,虑,12,月,月-2212,月,月-22,Thursday,December29,2022,10,、阅读,一,一切好,书,书如同,和,和过去,最,最杰出,的,的人谈,话,话。
04:48:2004:48:2004:48,12/29/2022 4:48:20AM,11,、越,是,是没,有,有本,领,领的,就,就越,加,加自,命,命不,凡,凡12,月,月-2204:48:2004:48,Dec-2229-Dec-22,12,、越是无,能,能的人，,越,越喜欢挑,剔,剔别人的,错,错儿04:48:2004:48:2004:48,Thursday, December 29, 2022,13,、知人者,智,智，自知,者,者明胜,人,人者有力,，,，自胜者,强,强12月-2212月-2204:48:2004:48:20,December29,2022,14,、意,志,志坚,强,强的,人,人能,把,把世,界,界放,在,在手,中,中像,泥,泥块,一,一样,任,任意,揉,揉捏,29,十,十,二,二月20224:48:20,上,上午04:48:2012,月,月-22,15,、,最,最,具,具,挑,挑,战,战,性,性,的,的,挑,挑,战,战,莫,莫,过,过,于,于,提,提,升,升,自,自,我,我,十,二,二,月,月224:48,上,上,午,午12,月,月-2204:48,December29,2022,16,、,业,业,余,余,生,生,活,活,要,要,有,有,意,意,义,义,，,，,不,不,要,要,越,越,轨,轨,。
2022/12/294:48:2004:48:20,29December2022,17,、一个,人,人即使,已,已登上,顶,顶峰，,也,也仍要,自,自强不,息,息4:48:20 上,午,午4:48 上,午,午04:48:2012月-22,,MOMODA POWERPOINT,Lorem ipsum dolor sit amet, consectetur adipiscing elit. Fusce id urna blandit, eleifend nulla ac, fringilla purus. Nulla iaculis tempor felis ut cursus.,感谢您,的,的下载,观,观看,专家告,诉,诉,。
