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TRANSCRIPT
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Dr. Alvin Fox
Nutrition, Growth and Metabolism
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KEY TERMS Obligate aerobeObligate aerobeObligate anaerobeObligate anaerobeAerotolerant anaerobeAerotolerant anaerobeFacultative anaerobeFacultative anaerobeMicroaerophilicMicroaerophilicSiderophoreSiderophoreMesophileMesophileThermophileThermophilePsychrophilePsychrophileGeneration timeGeneration time
Growth curveGrowth curveGlycolysisGlycolysisFermentationFermentationAnaerobic respirationAnaerobic respirationAerobic respirationAerobic respirationTricarboxylic acid Tricarboxylic acid (TCA) (TCA) cycle or Krebs cyclecycle or Krebs cycleOxidative Oxidative phosphorylationphosphorylation UbiquinoneUbiquinoneGlyoxylate pathwayGlyoxylate pathway
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Bacterial requirements for growthBacterial requirements for growth
• oxygen (or absence)oxygen (or absence)• energy energy • nutrientsnutrients• optimal temperatureoptimal temperature• optimal pH optimal pH
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Obligate aerobesObligate aerobes
• grow in presence of oxygengrow in presence of oxygen• no fermentationno fermentation• oxidative phosphorylationoxidative phosphorylation
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• no oxidative phosphorylation• fermentation• killed by oxygen• lack certain enzymes:
superoxide dismutase O2
-+2H+ H2O2
catalase H2O2 H20 + O2
peroxidase H2O2 H20 /NAD NADH
Obligate anaerobesObligate anaerobes
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Aerotolerant anaerobesAerotolerant anaerobes
• respire anaerobicallyrespire anaerobically
• not killed by oxygennot killed by oxygen
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Facultative anaerobesFacultative anaerobes
• fermentation fermentation
• aerobic respirationaerobic respiration
• survive in oxygensurvive in oxygen
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Microaerophilic bacteriaMicroaerophilic bacteria
• grow grow – low oxygenlow oxygen
• killed killed – high oxygenhigh oxygen
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Optimal growth temperature Optimal growth temperature
• Mesophiles: Mesophiles:
– human body temperaturehuman body temperature
* pathogens pathogens
* opportunistsopportunists
• pyschrophilepyschrophile– close to freezing close to freezing
• thermophilethermophile– close to boilingclose to boiling
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pH
• Many grow best at neutral pH
• Some can survive/grow
- acid
- alkali
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Nutrient RequirementsNutrient Requirements
• Carbon Carbon • NitrogenNitrogen• PhosphorusPhosphorus• SulfurSulfur• Metal ions (e.g. iron)Metal ions (e.g. iron)
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Siderophores (S)Siderophores (S)
Fe Fe 2+2+//SS
ReceptorReceptor
Fe Fe 2+2+//SS
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Measuring bacterial mass (live + dead) Measuring bacterial mass (live + dead) in liquid culturein liquid culture
TurbidityTurbidity(Cloudiness)(Cloudiness)
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Measuring viable bacteria
colonycolony
Colony forming unitsColony forming units
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Growth Curve
COLONY FORMING UNITS
TIME
Lag
Log
Stationary
Death
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Growth Curve
TURBIDITY(cloudiness)
TIME
Lag
Log
Stationary
Autolysis
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Generation timeGeneration time
• time for bacterial mass to doubletime for bacterial mass to double
• Example Example
100 bacteria present at time 0 100 bacteria present at time 0
If generation time is 2 hrIf generation time is 2 hr
After 8 hr mass = 100 x 2After 8 hr mass = 100 x 244
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SUGAR CATABOLISM
• Glycolysis Glycolysis – Embden Meyerhof Parnas PathwayEmbden Meyerhof Parnas Pathway
– most bacteriamost bacteria
– also animals and plants also animals and plants
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Other pathways for catabolizing sugars
• Pentose phosphate pathway (hexose Pentose phosphate pathway (hexose monophosphate shunt)monophosphate shunt)– generates NADPHgenerates NADPH– common in plants and animalscommon in plants and animals
• Entner Doudoroff Pathway Entner Doudoroff Pathway – a few bacterial speciesa few bacterial species
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•
GlycolysisGlycolysis
NADNAD NADHNADH
GlucoseGlucose PyruvatePyruvateC6C6 C3C3
ADP ADP ATPATP
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FermentationFermentation
PyruvatePyruvate
(C3)(C3)
NADHNADH NADNAD
Short chain alcoholsShort chain alcohols, , fatty acidsfatty acids(C2-C4)(C2-C4)
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Anaerobic Respiration = Anaerobic Respiration = Glycolysis + FermentationGlycolysis + Fermentation
NADNAD NADHNADH
NADHNADH NADNAD
ATPATP
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Krebs Cycle (C4-C6 intermediate compoundsKrebs Cycle (C4-C6 intermediate compounds)
PyruvatePyruvate 3CO3CO22
(C3)(C3)
NADNAD NADHNADH
NADHNADH NADNAD
Oxidative phosphorylationOxidative phosphorylation
OO22 HH22OO
ADPADP ATPATP
(C1)(C1)
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Aerobic Respiration =Aerobic Respiration =Glycolysis + Glycolysis +
Krebs Cycle/oxidative phosphorylationKrebs Cycle/oxidative phosphorylation
• Pyruvate to COPyruvate to CO22
– NADNAD toto NADHNADH
– glycolysis glycolysis
– Krebs cycleKrebs cycle
• Oxidative phosphorylationOxidative phosphorylation
– NADHNADH to to NAD NAD
– ADPADP to to ATP ATP
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Oxidative phosphylationOxidative phosphylation
• converts Oconverts O22 to H to H220 0 (oxidative)(oxidative)
• converts ADP to ATP converts ADP to ATP (phosphorylation)(phosphorylation)
• electron transport chainelectron transport chain
• ubiquinones/cytochrome intermediates ubiquinones/cytochrome intermediates
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The Krebs cycle
Citrate
Isocitrate
Alpha-keto glutarate
Succinate
Fumarate
Malate
Oxaloacetate
Pyruvate
-CO-CO22
Acetate
+-CO-CO2 2 NADHNADH
-CO-CO2 2 NADHNADH
C2
C
C
C4
X
x
C6
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Krebs Cycle - sugar as sole Krebs Cycle - sugar as sole carbon sourcecarbon source
PyruvatePyruvate
AcetateAcetate-CO-CO22
C4C4
PyruvatePyruvate+ CO+ CO22
+Citrate
CCC3C3
Oxaloacetate
Oxaloacetate
-2CO-2CO22
Aspartic acidAspartic acid
Krebs Krebs cyclecycle
ENERGYSTORAGE
BIOSYNTHESISC3C3
CCC2C2
C6C6C4C4
OxaloacetateX
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Krebs Cycle – fatty acids as Krebs Cycle – fatty acids as sole carbon sourcesole carbon source
Fatty acidsFatty acids
AcetateAcetate
+ CitrateOxaloacetate
-2CO-2CO22
Aspartic acidAspartic acid
Krebs Krebs cyclecycle
ENERGY
BIOSYNTHESIS
Isocitrate Succinate Glyoxylate+
AcetateAcetate+
MalateMalate
Oxaloacetatex
C4
C2
C2
C4C6
-2CO-2CO22
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The Glyoxylate and Krebs cycles
Citrate
Isocitrate
Alpha-keto glutarate
Succinate
Fumarate
Glyoxylate
AcetateMalate
Oxaloacetate
1
2
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Krebs CycleKrebs Cycle
– biosyntheticbiosynthetic
– energy storage energy storage
• Removal of intermediatesRemoval of intermediates
– must be replenishedmust be replenished
• Unique enzymatic replenishment pathwaysUnique enzymatic replenishment pathways
– sugars sugars
– fatty acidsfatty acids