Nutrition, Growth and Metabolism

Dr. Alvin Fox

1
 KEY TERMS
Obligate aerobe Growth curve
Obligate anaerobe Glycolysis
Aerotolerant anaerobe Fermentation
Facultative anaerobe Anaerobic respiration
Microaerophilic Aerobic respiration
Siderophore Tricarboxylic acid
Mesophile (TCA)
Thermophile cycle or Krebs cycle
Psychrophile Oxidative
Generation time phosphorylation
Ubiquinone
Glyoxylate pathway 2
Bacterial requirements for growth
• oxygen (or absence)
• energy
• nutrients
• optimal temperature
• optimal pH

3
 Obligate aerobes

• grow in presence of oxygen

• no fermentation
• oxidative phosphorylation

4
 Obligate anaerobes
• no oxidative phosphorylation
• fermentation
• killed by oxygen
• lack certain enzymes:
superoxide dismutase
O2-+2H+ H2O2
catalase
H2O2 H20 + O2
peroxidase
H2O2 + NADH + H+ 2H20 + NAD

5
 Aerotolerant anaerobes

• respire anaerobically
• not killed by oxygen

6
 Facultative anaerobes
• fermentation
• aerobic respiration
• survive in oxygen

7
Microaerophilic bacteria

• grow
– low oxygen
• killed
– high oxygen

8
 Optimal growth temperature
• Mesophiles:
– human body temperature
* pathogens
* opportunists
• pyschrophile
– close to freezing
• thermophile
– close to boiling

9
 pH
• Many grow best at neutral pH
• Some can survive/grow
- acid
- alkali

10
Nutrient Requirements
• Carbon
• Nitrogen
• Phosphorus
• Sulfur
• Metal ions (e.g. iron)

11
Siderophores (S)
Receptor

Fe 2+/S

Fe 2+/S

12
Measuring bacterial mass (live + dead)
in liquid culture

Turbidity
(Cloudiness)

13
Measuring viable bacteria
Colony forming units

colony

14
 Growth Curve

COLONY Stationary
FORMING
UNITS
Death
Log

Lag

TIME
15
 Growth Curve

TURBIDITY Stationary
(cloudiness)

Log Autolysis

Lag

TIME
16
Generation time
• time for bacterial mass to double

• Example
100 bacteria present at time 0
If generation time is 2 hr
After 8 hr mass = 100 x 24

17
 SUGAR CATABOLISM
• Glycolysis
– Embden Meyerhof Parnas Pathway
– most bacteria
– also animals and plants

18
 Other pathways for catabolizing
sugars
• Pentose phosphate pathway (hexose
monophosphate shunt)
– generates NADPH
– common in plants and animals

• Entner Doudoroff Pathway

– a few bacterial species

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 Glycolysis
NAD NADH
Glucose Pyruvate
C6 C3
ADP ATP

20
 Fermentation

NADH NAD
Pyruvate Short chain alcohols,
fatty acids
(C3) (C2-C4)

21
Anaerobic Respiration =
Glycolysis + Fermentation
NAD NADH
ATP
NADH NAD

22
Krebs Cycle (C4-C6 intermediate compounds)
NAD NADH
Pyruvate 3CO2
(C3) (C1)

Oxidative phosphorylation
NADH NAD
O2 H2O
ADP ATP
23
 Aerobic Respiration =
Glycolysis +
Krebs Cycle/oxidative phosphorylation

• Pyruvate to CO2

– NAD to NADH
– glycolysis
– Krebs cycle
• Oxidative phosphorylation
– NADH to NAD
– ADP to ATP
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Oxidative phosphylation
• converts O2 to H20 (oxidative)
• converts ADP to ATP (phosphorylation)
• electron transport chain
• ubiquinones/cytochrome intermediates

25
 The Krebs cycle

C2 Isocitrate X
C
Acetate Citrate -CO2 NADH
C6
-CO2
+ Alpha-keto
Oxaloacetate C4 glutarate
Pyruvate -CO2 NADH

Malate x
Succinate C
Fumarate

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 Krebs Cycle - sugar as sole
carbon source
Krebs
Acetate +
-CO2 cycle
Oxalo
Pyruvate C2 C4 acetate Citrate
C C6
C3 -2CO2
BIOSYNTHESIS ENERGY
STORAGE
X
Oxalo
acetate
Aspartic acid

+ CO2
Pyruvate Oxaloacetate
C3 C C4

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Krebs Cycle – fatty acids as
sole carbon source
ENERGY
Acetate Krebs
Fatty acids + Oxalo cycle
Citrate
acetate
BIOSYNTHESIS -2CO2

x
Oxalo
acetate Aspartic acid

C2
Isocitrate Succinate + Glyoxylate C2
-2CO2
+ Acetate
C6 Krebs cycle C4
Malate C4
28
The Glyoxylate and Krebs cycles

Isocitrate
Citrate
1 Alpha-keto
Oxaloacetate Glyoxylate glutarate
2
+ Acetate
Malate Succinate
Fumarate

Krebs and Glyoxylate cycles

Krebs cycle only Glyoxylate cycle only
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 Krebs Cycle
– biosynthetic
– energy storage

• Removal of intermediates
– must be replenished
• Unique enzymatic replenishment pathways
– sugars
– fatty acids 30