15 英文版PowerPoint课件 Chapter 5 Microbial Nutrition and Culture Medium.ppt

第1页：Chapter 5 Microbial Nutrition and Culture Medium

Chapter 5 Microbial Nutrition and Culture medium

第2页：Nutrition：Nutritionisaprocessbywhichchemicalsubstancescallednutrientsarerequired

Nutrition：Nutritionisaprocessbywhichchemicalsubstancescallednutrientsarerequiredfromtheenvironmentandusedforthecellularactivitiesofmetabolismandgrowth.Nutrient：the substance could be used by microbes for metabolism or reproduction of cells.

第3页：Nutrient provide the

Nutrient provide the structural substance , energy,metabolic regulate substance and appropriatephysiological environment.Some microbes could use nonmaterial energy---light.

第4页：I.cellular chemical

I.cellular chemical constitutesIt’s much alike in the life in the element . The main elements are C、H、O、N (90%---97% to total dry cell weight), C is about 50%, the others are inorganic nutrient. C/N is 5:1 usually.Section 1 Six kinds of essential nutrient for microbes

第5页：elementØMacronutrient：C、H、O、N、P、SØothers

elementØMacronutrient：C、H、O、N、P、SØothers element：K、Na、Ca、Mg、Fe、Mn、Cu、Co、Zn、Mo,et alExistence form:Øorganic：protein、sugar、lipoid、nucleic acid、vitamin、decompositionsubstance、metabolic medial productsØinorganic salt ashØWater---70% ~ 90% to wet cellweight

第6页：elementbacteriayeastmouldC5049.847.0N157.55.2H85.76.7O2031.140.2P31.51.2S10.30.2

elementbacteriayeastmouldC5049.847.0N157.55.2H85.76.7O2031.140.2P31.51.2S10.30.2Table1.Themacronutrientcontentinmicrobes(%todrycellweight)

第7页：The substance utilized

The substance utilized by generic species could be utilized by microbes too.The substance couldn’t be utilized by generic species could be utilized by microbes.Some kinds of substance harmful to generic species also could be utilized by microbes.Microbes is omnivorous.

第8页：II. Main nutrient

II. Main nutrient and the functionCarbon sourceNitrogen sourceEnergy sourceGrowth factorInorganic saltWater

第9页：Functions:vconstitute

Functions:vconstitute the cells of the microbesvprovide the energy needed in thephysiological activity to microbesvthe source to form the microbialmetabolic productsNutrient is the substance base formicrobial metabolism and living activity.If lose this base, the life will die.

第10页：Heterotrophicmicrobe

• Heterotrophicmicrobe:isanorganismmustobtainitscarboninanorganicform.Autotrophicmicrobe:isanorganismthatusesCO2,aninorganicgas,asitscarbonsource.1.Carbon sourcesThe nutrient provide the carbon element for microbies
• Organic carbon sources: protein, nucleic acid, starch, glucose, et al
• Inorganic acid: CO2 ,Na2CO3, CaCO3 et al

第11页：the carbon sources

the carbon sources could be utilized by microbes(compound)saccharide：glucose，fructose，maltose，sucrose，starch，galactose，lactose，mannancellulose，hemicellulose，lignin，et alorganic acid：lactic acid，citric acid，，short chainfatty acid，

第12页：organic nitrogen sources：peptone、soybean

• organic nitrogen sources：peptone、soybean powder、maize slurry
• inorganic nitrogen sources：NH4NO3、(NH4)2SO4
• gas nitrogen sources：N2 in the atmosphere2.Nitrogen sourcesThe nutrient provide the nitrogen element for microbes.Normally,nitrogen source couldn’t be used as energy sources.
• quick nitrogen sources
• slow nitrogen sources

第13页：energysourcesarethesubstancesorradiantenergythatcouldprovideprimalenergysources

energysourcesarethesubstancesorradiantenergythatcouldprovideprimalenergysources.Heterotrophs’scarbonsourcescouldalsobeitsenergysources.3. Energy sourcesInorganic:energy to chemoautotrophsEnergy sources tableChemical substancesRadiant energy：energy to photoautotrophs and photoheterotrophs.Organic: energy to chemoheterotrophs

第14页：mono-function:radiantenergydi-function:reductiveinorganicnutrient,suchasNH4+isth

mono-function:radiantenergy

• di-function:reductiveinorganicnutrient,suchasNH4+istheenergysourcesaswellasthenitrogensourcesofnitratebacterial.
• triplefunction:thenutrientsincludingN、C、H、Oareenergysources、carbonsourcesandnitrogensourcesheterotrophsmicrobes.one kind of nutrient could provide more than one kind of function

第15页：Definition的概念

• Definition:An organic compound such as an amino acid, nitrogen base, or vitamin that cannot be synthesized by an organism and must be provided as a nutrient is a growth factor.Sources of growth factors in culture medium:yeast extract、maize slurry、wort,et al。
• 4.Growth factorFunction：coenzyme or necessary to activation of enzyme.

第16页：narrow sense：vitaminbroad

• narrow sense：vitaminbroad sense：vitamin, amino acid, base, fatty acid ,et al
• auxoautotrophs
• auxoheterotrophs
• nutritional deficiency mutant
• auxo-excessivetrophs

第17页：vitaminspeciesthiamin

vitaminspeciesthiamin B1Bacillus anthracislactoflavinClostridium tetaninicotinic acidBrucella abortusvitamin B6Lactobacillus spp.biotinLeuconostoc mesenteroidespantothenic acidProteus morganiifolacinLeuconostoc dextranicumB12Lactobacillus spp.vitamin KBacteroides melaninogenicusVitamin necessary to some bacterials

第18页：5. Inorganic saltsMacroelement

5. Inorganic saltsMacroelement: need the concentration of the element at 10-3-10-4M Microelement: need the concentration of the element at 10-6-10-8M

第19页：The physiology function

The physiology function forinorganic saltsinorganic saltsMacroelementMicroelementUsual functionEspecial functionNormal molecular in cell(P、S、Ca、Mg、Fe)Physiology modulation materialmaintain osmotic pressure (Na+) enzyme activator(Mg2+) stablize pHEnergy sourse of chemoautotrophic bacteria(S、Fe2+、NH4+、MO2-)Hydrogen aceptor during respiration without oxygen (NO3-、SO42-)Enzyme activator (Cu2+、Mn2+、Zn2+)Molecular of special structure (Co、Mo)

第20页：Physiologicalfunction：cellcompositionsolventofbiochemistryreactionchemicalandphy

Physiologicalfunction：

• cellcomposition
• solventofbiochemistryreaction
• chemicalandphysiologicalreactionmedium
• materialtransferredmedium
• regulatecelltemperature
• maintainosmoticpressure6.Waterexisted state:dissociated state(solvent)and integrated state(structure composition)

第21页：◆based on different

• based on different carbon sources：heterotrophs: can’t use CO2as its mainly or sole carbon energy autotrophs: can use CO2as its mainly or sole carbon energy Section 2 Nutrition types of microbes
• based on different energy sources ：phototrophs: obtain energy from lightchemotrophs:obtain energy by oxidation
• basedongrowthfactor：prototrophorwildtypeauxotrophClassification

第22页：Nutrition type of

Nutrition type of microbeNutritiontypeEnergysourceCarbonsourceexamplephotoautotrophssunlightCO2Cyanobacterial，Purplebacterial，Greensulfurbacterial，AlgaephotoheterotrophssunlightCO2andsimpleorganiccompoundsRhodospirillumchemoautotrophsinoganiccompoundsCO2NitrobacterialSulfbacterialIronbacterialHydrobacterialchemoheterotrophsorganiccompoundsorganiccompoundsMostmicrobes，Prozotoa

第23页：parasitism ——live in

• parasitism ——live in live organisms.
• saprophytism——live in dead organic.Chemoheterotrophsto note that such classification is not absolute，and it may be changed under different conditions。
• heterotrophs：aredependentonotherlifeforms，canlivenormallywithatleastonekindoforganiccompound，itscarbonsouecesandhydrogensuppliershouldbeinorganicform，energycanbeobtainedbyoxidationandabsorptionofsunlight.
• autotrophs：can live normally without any organic compounds.

第24页：Section 3 Transport

Section 3 Transport mechanisms for nutrient absorption Theabsorbtionofnutrientandexcretionofmetabolizedproductsrefertotransportionofmolecular、nutrientabsorbedincellandbeused、metabolizedproductssecretedoutofthecell,thatisthewholeprocessoftransportion.The cell wall can only block large molecular solute (>600Da) out of the cell,while the membrane which have lipid bilayer and integral protein is the main barrier to control the nutrient’s flow.

第25页：permeability is different

• permeability is different from absorbtion
• commonly,large molecular first hydrolyze to small molecular,then be aborbed.
• fat-dissolved：easily permeable
• ionization：the weaker of polar,the faster of the permeable

第26页：I. Simple diffusionThe

I. Simple diffusionThe rate of diffusion is directly proportionalto the concentration gradient of the moleculeacross the membrane.；Do not acquire metabolic energy, is of nospecificity.Only transformed small molecular,O2、CO2、glycerol、ethanol and some amino acids；Lipohilia molecules make use of thepermeability of cell membrane to transport

第27页：II. Facilitated diffusionThe

II. Facilitated diffusionThe transported molecular in and out ofthe cell have different appetency to thecarrier protein.characteristic：Require the presence of specificcarrier protein, down the concentrationgradientDo not acquire metabolic energyTransport SO4 , PO3, sugars

第28页：Carrier protein:namely

Carrier protein:namely permeable enzyme（mostly are induction-enzyme），is specific for one particular molecular or a group of structurally similar moleculars. It can make the transportion faster,but is not able to make it against the concentration gradient.

第29页：facilitated diffusionextracellularmembraneintracellular

facilitated diffusionextracellularmembraneintracellular

第30页：单纯扩散促进扩散Concentration

单纯扩散促进扩散Concentration gradientTransportion ratePrapartion of simple diffusionand facilitated diffusion

第31页：simple diffusion

• simple diffusion 、facilitated diffusion and active transport：the transported solute won’t be changed.III.Active transportcharacteristics：
• The main mode for microbes to absorb nutrient
• Against the concentration gradient ,require energy
• Need barrier protein, is specific
• Transport organic iron、inorgnic iron、amino acid、lactose and other sugars
• Need energy to change the conformation of barrier protein
• Appetence change protein conformational changeconsume energy

第32页：intracellularExtracellular(outsidr

• intracellularExtracellular(outsidr the plasma membrane)election transfer1. Use electron transfer to pump the proton to extracellular2. Expel Na+ out of membrane by reverse transport mechanism3. Na+ combine with barrier protein complex4. The form of combination site changed,to combine with the solute(eg:sugar,amino acid)
• 5. Barrier protein conformational change, release Na+ in theintrecellular,then solute separate from barrier protein. Mechanism of active transport：use the gradient of H+ and Na+

第33页：characteristics：belong

characteristics：

• belong to active transport
• solute molecular have chem-modification
• directional phosphorylation
• need a complicated series of transport enzyme
• to transform glucose、fructose、mannan、purin、nucleotide、fatty acid,et al IV. Group translocationThe membrane has high impermeability to most phosphate compounds. Input each glucose may consume one ATP.

第34页：Mainly depend on PEPand

• Mainly depend on PEPand PTS.PEP+ HPrEIPyruvic acid+ P-HPrP-HPr +saccharideEIIsaccharide-P+HPr1. Activate the heat stable carrier protein (HPr)
• 2. Saccharide transferred into membrane after phosphorylation

第35页：Transport of glucose

Transport of glucose grouptranslocation2kindsofsystemsofPEPandPTSsystem.High-energyphosphatetransferfromHPrtodissolvedEIIA,EIIAcombineswithEIIBmannantransportsystemandseparateintheglucosetransportsystem.Nomatterinwhichform,phosphatewillfirsttransferfromEIIAtoEIIB,andarriveatglycosylbytheprocessoftransportacrossmembrane.cytoplastCytoplasm stroma

第36页：I. Definition:medium，culture

I. Definition:medium，culture medium：Culture medium is the mixed nutrient suitable for microbe’s growth and breed.Section 4 culture medium

第37页：A. Four principles1.

A. Four principles1. Definite aims（different microbes have different nutrient requirement）:to culture which microbes，to get which products，uselessII. Principles and methods to select and make culture medium2.Suitable nutrients:pay attention to the concentration and ratio of nutrients，especially C/N ratio

第38页：C/Nratio:molratioofcarbonatomincarbonsourcesandnitrogenatominnitrogensources.C/N

C/Nratio:molratioofcarbonatomincarbonsourcesandnitrogenatominnitrogensources.C/Nratio in most medium:100/0.5～2 .Nitrogen content %：NH3(82%)>CO(NH2)2(46%)>NH4NO3(35%)>(NH4)2CO3(29.2%)>(NH4)2SO4(21%)

第39页：Selection of suitable

Selection of suitable nutrientkindexampleexplaiationmonosaccharidesglucose,maltose,xylose,galactose,fructoseusedasquickcarbonsources,butwillspeedupbreathofmicrobe,decreaseresolvedO2,makemid-metabolizedproductsaccumulate.disaccharidesaccharose,lactoseUsedasslowcarbonsources,butwillspeedupbreathofmicrobe,decreaseresolvedO2,makemid-metabolizedproductsaccumulate.polysaccharidestarch,cornmeal,celluloseslowcarbonsources,onlyfewmicrobescanusecelluloseascarbonsourcesmolassesSugarbeetmolasses,sugancanemolasses.goodcarbonsources,complicated,usuallyfermentedfromyeastinfermentationindustry,widelyusedintheproductionofantibiotic、acetoneandbutanol.usuallyincludesaccharose35%,glucose9%,fructose7%,8kingsothersugars4%,otherdeoxidizedmaterials3%,24kindsaminoacid4.5%,organicacid5%,wax、steroidsandphosphorous0.4%,ash12%,water20%.maltbuddedbarleymainlyusedtocultureyeast,canbeusedtoprouducebeerinindustrygreasecottonseedoil,cornoil,beanoil,sunflowerseedoilhigh热值,consumemoreO2whenculturedinliquid,usuallyaddedas消泡剂inindustryalcoholsmethanol,glycerolMethanolcanbeusedtocultureyeasthydrocarbonCH4,C2H4,C4H10,C12~C20high热值,consumemoreO2whenculturedinliquid.hardlydissolved,don’ttoomuchbeatingupwhenculturedinliquid.organicacidaceticacid,lacticacid,citricacidwillincreasethepH

第40页：carbonsourcesstromaglucosemethaneNormalparaffinhydrocarbonmethanolethanolacetate

carbonsourcesstromaglucosemethaneNormalparaffinhydrocarbonmethanolethanolacetatefumaratecellproducingefficiency(gcell/gstroma)0.510.621.030.400.680.340.36Cell yield by different energy source

第41页：CommonnitrogensourcesformicrobeKindsSuitablemicrobeCharacteristicorganic:beafext

CommonnitrogensourcesformicrobeKindsSuitablemicrobeCharacteristicorganic:beafextract,yeastextract,peanutcakepower,soybeanpoder,cottoncakepower,maizeslurry,maizeproteinpower,peptone,yeastpower,fishmeal,silkwormpupapower,wheatbranMostmicrobes,eg:Penicillium,yeast,E.colicontainplentyofaminoacid,goodnitrogensourcesformicrobesinorganic:ammoniumsalt，nitrate,ammoniaPartofmicrobes(aminoacidheterotrophsaren’tabletouseit)Nitratecanbeusedafterdeoxidization,mayregulatepHmeantime.N2fewbacteria(approximately50genus,100species)Nitrogenfixationissecondimportantonlytophotosynthesis.

第42页：pHosmotic pressure

pH

• osmotic pressure and water ability
• redox potential3.Suitable chemical and physical conditions:

第43页：pH：most suitable pHvbacteria

pH：most suitable pHvbacteria: pH7.0~8.0vactinomyces：pH7.0 ~8.5vyeast: pH3.8~6.0vmould：pH4.0~6.0pH value in the medium could change in the growing processof microorganisms：vIf culture in stroma containing carbohydrate,produce acid,sothe pH will decrease .vIf decompose protein and amino acid, produce NH3,pH will

第44页：methods to maintain

methods to maintain mediumpHstableUse phosphorus buffer：K2HPO4/Na2HPO4, H2PO4/NaH2PO4Use“standby alkali”：CaCO3、CaHCO3Use acidesecent salt：citrate、lactophosphate et alUse liquid N2 or HCl.

第45页：Osmotic pressure:pressure

Osmotic pressure:pressure produced by solutes, when filterring from high concentration to low cocentration。

Hypertonic solution will cause plasmolysis.Hypotonic solution will make cell expand or even break.

第46页：Utilized ability of

Utilized ability of waterwater activity, awaw：the quantity of free or dissociated water thatmicrobes can utilize under natural environment.It’s eaqual to the ratio of saturated steam pressureof the solution and pure water, in the sametemperature and pressure..aw =P/PoP: steam pressure of the solutionPo: steam pressure of pure water

第47页：solutionaw30% glucose0.9641%

solutionaw30% glucose0.9641% glucose +20%glycerol0.9551% glucose +40%saccharose0.964Saturated NaCl0.78Saturated CaCl20.30Saturated MgCl20.30Saturated LiCl0.11Aw of some solution

第48页：Eh（redox potential）：to

• Eh（redox potential）：to show the trend of reducer releasing electron or oxidant accepting eletron in the redox system.
• Aerobes：＞+ 0.1V。Generally +0.3~ +0.4V
• Anaerobes：< + 0.1
• Facultative anaerobes：< +0.1 fermentation>+0.1 aerobic respiration

第49页：4. Appropriate raw

4. Appropriate raw materialveconomy principle:use the low gradereplacing high grade,use the wastereplacing normal material,use thesimple replacing complexveasily treated,with low costvproduce little waste after treated

第50页：SterilizationSterilization

SterilizationSterilization under high pressure：1.05kg/cm，1210C，15～30minSterilization under high temperaturemay destroy some nutrients andchange the pH.

第51页：B. Four methodsEcological

B. Four methods

• Ecological simulation
• Lookup literature
• Elaborate
• Experiment Comparison

第52页：Category of Culture

Category of Culture medium Based on the component of the culture medium：Complex medium ( or Undefined medium ) Complex mediumis one kind of culture mediums, which utilizes animal/ plant/micro-organism or the extract from these substance, and the components are undefined, e.g. broth peptone medium ,which is a growth medium for bacteria; malt extract medium ,which is a growth medium for yeast.Advantages：convenience;berichinnutrition;variety

第53页：Definedmedium(orSyntheticmedium)Definedmediumisonekindoftheculturemediums,whichi

Definedmedium(orSyntheticmedium)Definedmediumisonekindoftheculturemediums,whichiscompoundedbymanypurechemicalreagent,andthecomponents(includingthemicroelement)aredefinedexactly,e.g.Glu-NH4+medium,whichisagrowthmediumforbacteria;Starch-NO3-medium,whichisagrowthmediumforactinomycete.Expensive，beusedinresearch(metabolismanalysis/geneticanalysis)Advantages：Thecomponentisexact;Well-repeatSemi-defined medium

第54页：Solidmedium:addsolidifyingagents;thecontentofsolidifyingagentsis1～2％.Conditionof

• Solidmedium:addsolidifyingagents;thecontentofsolidifyingagentsis1～2％.Conditionofbeingthesolidifyingagents:Can’tbeutilizedordecomposedbymicrobe;cankeepthesolidconditioninthegrowthtemperaturerange;Itisn’tharmfulformicrobeatsolidificationpointtemperature;can’tbedestoriedbysterilization;good-diaphaneity;convenience;cheap.Solidifyingagentsthatwealwaysuse:AgarandGlutinUsage:isolation/identification/conservationofthestrain(Solidmediumallowsthesurfacetoformmono-colony)
• Base on the physical condition of culture medium:

第55页：Semi-solid medium:the

Semi-solid medium:the content of solidifyingagentsis 0.5%Usage: observe the motion of bacteria;mensuratethe titer ofbacteriophage.Liquid medium :no solidifying agentUsage: microbe culture；the research on physiology

第56页：The Difference between

The Difference between Agar and GlutinChemistryComponentNourish-mentResolv-abilityMelttemperatureSolidificationtemperatureConcent-rationDiaphaneityAdhesion valueTolerance ofSterilizationunderhighpressureAgar

第57页：E.ColigrowedinEMB(EosinMethyleneBlue)Culture,candecomposelactoseandproducemuchmi

E.ColigrowedinEMB(EosinMethyleneBlue)Culture,candecomposelactoseandproducemuchmixedacid.ThethalluscarrywithH+,soacidicchromophoreEosinisattractedtothepositivelychargedmoleculesofE.coli.EosincancombinewithMethyleneBlue,whichmakesthecoloniespurple.Greenmetalliclusterisobservedfromthesurfaceglistenofcolonies.Coloniesthatfermentlactosearered，andcoloniesthatproduceacidweaklyarebrown,whereasnon-fermentersaretransparent.EMB (Eosin Methylene Blue) Culture medium identified enteric bacterial

第58页：EMB (Eosin Methylene

EMB (Eosin Methylene Blue) Culture medium

第59页：Brief Summary1、Microorganisms

Brief Summary1、Microorganisms need about 10 kinds of macroelement，part ofthem are used to form carbohydrate、lipid、protein、nucleicacid,others are bittly used as coenzyme and prosthetic group.2、All the microorganisms can be classifed on the basis of theirneedness of carbon courses、energy sources

第60页：Reviewing question

• Reviewing question of this chapter1、What are the macroelement and microelement?
• 2、How manykinds of macroelements are needed by microbes?3、What is nutrition,what is nutrient,What physiological functiondoes the nutrient have？4、Try to compare the similarities and differences about nutrients

第61页：8、What are the characteristic

8、What are the characteristic of the using of nitrogensources by microbes？9、What is amino acid autotrophs,what is amino acidheterotrophs？10、Try to compare the differences of the way thatnutrients use to get into cell.11、What is culture medium and what is complexmedium？12、What is semi-solid medium？

第62页：References：Zhou Deqin，Microbiological

References：Zhou Deqin，Microbiological Tutorial，2002, second edition，higher education pressKathleen Park Talaro,fundations in microbiology,2005, fifth edition, higher education press