生物分离工程-双语版 2

Chapter2Stepsinbioseparationprocesses1IntroductionBasicallythreestepsinbioseparationprocess(1)Celldisruption(2)Initialfractionation(3)High-resolutionfractionationEffectiveseparationprocessesforgeneticengineering:(1)Stable(2)Pure(3)Homogeneous(4)Activeform(5)Highyield1IntroductionConsideringproteinpropertiesforbioseparation(1)size(2)charge(3)biologicalaffinity(4)hydrophobicity-solubilityProductsfrommammaliancelltechniques(1)enzyme(2)hormones(3)nonantibodyimmunoregulators(4)monoclonalantibodies(5)tumorspecificantigens1IntroductionProteinfolding(1)shortrangedinteraction(secondarystructure)(2)gradualcombinationandreshuffling(重新组合）(subdomains,domains,andsubunitassemblies)Secondgenerationbiotechnologicalproducts(1)largermolecules(2)morecomplex(3)moreglycosylate(4)moresulphydrylbonds1IntroductionInitialbioseparationimportance(1)filtration(2)centrifugation(3)chromatographyClassifiedheuristics（论据，规则）(1)unitoperationmethods(2)designstepsequence(3)species(componentsproperties)(4)composition1IntroductionThreefactorsofdesigningbioseparationprocesses(1)purity(2)cost(3)marketMaterialstateandchoiceofseparationmethodsMaterialsecreted(ultrafiltration,centrifugation)Notsecretedmaterial(celldisruption,solid-liquidseparation)Materialinliquid(ultrafiltration,adsorption)Materialinsolid(extractionintoaqueoussolution)NewandwordsProtactelawsuits诉讼Deleterious有害的，有毒的Vexing烦恼Hierarchic分等级Subdomain亚域Domains蛋白质结构的域Subunit亚单位Endoplasmicreticulum内质网Disulfidebond二硫键Academia学术界Delicate脆弱的Flowchart流程图Cytoplasmic细胞质的Periplasmic外周胞质Osmotic渗透urea脲Guanidine胍IIProductexcretionfromthecellorcelldisruptionExcretedproteinadvantage(1)contaminationlevellow(2)easydownstreamprocessingsteps(3)notrequirecelldisruption(4)avoidhandlingviscousfluidsandproteinaserelease(5)facilityextractionofproducts(6)signalpeptidedeterminessecretionCelldisruption(1)energy-intensive(强烈）andviolent(2)withoutdestroyinginterestprotein(3)contain(兼容）process(bacteria,yeast,mammalian,plant,andinsectsources)(4)becontrolled(5)bevalidatedProductsrequiringcelldisruptionVaccines(tetanus(破伤风）,meningitis（脑膜炎)Glucokinase（葡萄糖激酶）Glycerokinase（甘油激酶）Invertase（转化酶）Toxin(enterotoxin)Subcellularcomponents(mitochondrai,chloroplast)Intracellularconstituents(DNA,RNApreparation,andvirus-likeparticles)RecombinantinsulinRecombinantgrowthhormoneProteinAandG.Ideallargescalecelldisrupter(1)disrupttoughorganisms(2)mechanismofcelldisruptionshouldbewellunderstood(3)sterilizable(4)containable（兼容）andvalidatable(5)amenabletobeingmadeautomatic(6)continuousandcompact（紧凑的）(7)economicalCelldisruptiongoalObjectivefunction(monetary（金钱）valueofproduct)Smallscalecelldisruption(1)ultrasonic(2)freezethawing(熔化）(3)forcecellsthroughverysmallorificesathighpressure(4)glassorporcelainballmillsLargescalecelldisruptionHomogenizerStructure(positivedisplacementpump,pistons,nozzle)Mechanisms(1)Shear(2)Cavitation（气穴）(3)impingement(冲击）Condition(1)concentration(4-175gdrywt/liter)(2)pressure(30-95MPa)(3)passesnumber(~5time)(4)modeledbyequation(5)wild-typeE.colicellsmoredifficulttodisrupt(heatinductionat42C)Ln(1/(1-R))=KNPΓDelineatingmechanismsofcelldisruption(1)facilitatecontrolling(2)validating(3)reproducible(4)betterdesignandoptimizeequipment(5)Turbulenteddies(湍流涡旋）Responsibleforcelldisruptionoscillate)(6)shearstress,turbulencebyimpingementEnzymaticlysiscelldisruption(1)alyticprotease(dissolvetheouterpartofthecellwall(2)endo-β-(l-3)glucanase（葡聚糖酶）todisintergrateglucannet(3)source（酶的来源）andculturesystemkineticanalysisAunderstandingthesystemBdesigningthesmallandlargescalesystemsCcytophaga（吞噬细胞）systemexhibitshighproteaseactivity,higherconversionandhigherlysisrateatlongertimeDoerskoviasystemshighglucanaselowproteaseactivity,containsomeinhibitorsEaninitiallaginyeastlysisbybothenzymeFsimpletwostepmodel(cellmasssolubilized,proteinhydrolyzedtopeptides)GMishaelis-MentenkineticequationpredictcellwalllysisHproductionoffoodgradesinglecellproteinNewwordsandvocabularyAmylase淀粉酶Modulate调节Interferons干扰素Viscousfluids粘性流体Signalpeptide信号肽Carboxypeptidase羧肽酶Alkalinephosphatase碱性磷酸酯酶Glyceraldehyde-3-phosphatedehydrogenase甘油醛-3-磷酸脱氢酶Reducednicotinamideadenine还原型尼克酰胺腺嘌呤Dinucleotide二核苷Reductase还原酶Hydrophobic疏水的Hydrophilic亲水的Newwordsandvocabularies(2)Pathogens病源体Ballmills球磨机Resonance共鸣，回声Vibration振动，摆动Dissipate分散Pinpointing最佳点,针尖Oscillate振荡Candidautilis假丝酵母Extrusion喷出Sapphire兰宝石Jewel宝石Thermocouple热电偶Sleeve袖子，套Hydrodynamic流体力学Hydroxylaseenzyme羟化酶Synergistic增效，协同作用Glucan葡聚糖MorphologyIIIinitialfractionationPrimaryisolationstep(1)adsorption(2)extraction(3)precipitation(4)distillation(5)filtration(6)solventevaporationRequirementSignificantvolumereductionInterestproductinsolution(1)graduallyremoveimpurities(2)increaseproductconcentration(3)chosemethodsdependsonscaleandorganism(4)washingstagesfollowedbydewateringstepYeastseasytodealwith(1)10µmindiameter(2)easilyflocculated(3)centrifugation(4)filtration