1. Microbial Metabolism

2. Metabolism Relationships

3. Metabolism Pathways

4. Metabolism Define Requirements Rate Types Energy Enzymes Limiting step
Reaction time
Types
Anabolic
Endergonic
Dehydration
Biosynthetic
Catabolic
Exergonic
Hydrolytic
Degradative
+/- metabolites

5. Metabolic Diversity Energy generating metabolism
Fermentation
Alcohol
Acid Formation
Lactic Acid
Mixed Acids
Others
Respiration
Aerobic
Anaerobic
Biosynthesis of secondary metabolites
Heterotrophic
Autotrophic

6. Energy Forms Use Chemical Types Heat Kinetic Potential Chemical
Mechanical
Electrical
Radiation [EM]
Chemical Types
ATP
UTP
GTP
Heat
Byproduct
45%

7. Various Types of Prokaryotic Energy Production Processes
Fermentation
Anaerobic Respiration
Aerobic Respiration
Lithotrophy
Photoheterotrophy
Anoxygenic photosynthesis
Methanogenesis

8. Enzymes Structure Characteristic functions Protein Ribozyme [ribosome]
Active site
Specific
Modified Forms
Inactive
Active
Coenzyme/Cofactor
-ase
Polymerase
Others: Lyases, Hydrolases,
Isomerases, Transferases

9. Enzyme Characteristics

10. Enzyme Structure Allosteric site Apoenzyme Protein Allosteric site
Cofactor
Metal ions Cu Zn Mg Fe Ca Co Mn
Coenzyme
Vitamins
CoA
NAD
NADP
FAD
FMN
Create Holoenzyme with active site
Allosteric site

11. Factors Affecting Enzymes
Temperature pH
Acids/Bases
UV light
Concentration of substrates (saturation)
Inhibitors

12. Enzyme Inhibitors Classified Competitive Noncompetitive @ active site
Similar shape
Binding
Irreversible
Reversible
Noncompetitive
@ allosteric site
Change shape

13. Noncompetitive Enzyme Inhibition

14. Enzyme Inhibition Summary
Noncompetitive at
Allosteric site
Competitive
at active site
Excitatory
Inhibitory
Reversible
Irreversible

15. Glycolysis: Embden-Meyerhoff
Glycolytic
Cytoplasm
Anaerobic
End products
2 Pyruvic acids
4-2 = 2 net ATP by substrate level phosphorylation
2 NADH
2 H20

16. Lactic Acid Formation

17. Glycolysis: PPP Breakdown 5-6 C Cytoplasm Anaerobic End products 1 ATP
2 NADPH
CO2
4,5,6,7 C AA
Nucleotides
Glycolytic pathways
Photosynthesis

18. Glycolysis: Entner-Duodoroff [E-D]
Glycolytic
Cytoplasm
Anaerobic
Different enzymes
Pseudomonas
Enterococcus
End products
2-1 = 1 net ATP
NADPH
NADH
2 Pyruvic acids
H20
NADP+
NADPH

19. Anaerobic Pathways ComparedEM
PPP ED
Location
Substrate
Cytoplasm
Glucose
G6P
Steps
2 parts
6C  3C
Different sugars (4-7C)
Glyceraldehyde
Products
2 Pyruvate
2 ATP
2 NADH
2 H+
Pyruvate & Fructose
6 CO2
12 NADPH
12 H+
1 ATP
1 NADPH
1 NADH

20. Glycolytic Pathways used by various Bacteria
Bacterium
E-M
PPP
E-D
Acetobacter aceti - +
Bacillus subtilis
major
minor
E. coli
Lactobacillus acidophilus
Pseudomonas aeruginosa
Vibrio cholera

21. Anaerobic Processes Lactic Acid Mixed Acid Butanediol Butyric Acid
Lactobacillus
Mixed Acid
Enterobacteriaceae
Butanediol
Klebsiella
Enterobacter
Butyric Acid
Clostridia
Butanol-Acetone
Propionic Acid
Corynebacteria

22. Fermentation

23. Fermentation Pathways

24. Fermentation Summary Anaerobic Cytoplasm Partial Oxidation
Small amounts of ATP generated via substrate level phosphorylation
Organic intermediaries as final electron acceptors
End products
Acid: Lactic Acid, Acetic Acid, Butyric Acid, Acetone
Alcohol: Ethanol, Isopropyl
Gas : CO2, H2
Contaminants

25. Summary

26. Carbohydrate Fermentation Tests

27. Phosphorylation Substrate Level Oxidative Phosphorylation
Direct transfer of phosphate
Glycolysis
Oxidative Phosphorylation
Electron transfer
Chemiosmosis
Photophosphorylation
Light energy to chemical energy

28. Substrate Level Phosphorylation

29. Aerobic Respiration

30. Mitochondria of Eukaryotes

31. Plasma [cell] membrane

33. Acetyl CoA Formation

34. Acetyl CoA Final Structure

35. Krebs Cycle TCA/Citric Acid
Eukaryotic
Mitochondria Or
Prokaryotic
Cytoplasm

36. Oxidation in Krebs Cycle

37. Krebs Cycle Metabolites
For every AcetylCoA
2 CO2
3 NADH
1 FADH2
1 ATP (substrate level phosphorylation from GTP)
Regenerates
CoA
Oxaloacetic acid

38. Dehydrogenation Use of hydrogen in oxidative reactions
Removal of electron from hydrogen
Carried on vitamin B derivatives
Energy released is trapped in chemical bonds

39. Redox Reactions

40. Oxidation

41. Reduction

42. Catabolism + Anabolism

43. Eukaryotic ETC

44. Prokaryotic ETC Vit B2 Riboflavin based Flavoproteins
Metal Proteins with ions (Fe, S, Cu)
Iron based Cytochromes (b,c1,c,a,a3)
Ubiquiones based on Vit K (CoEnzyme Q)

45. ETC Steps Electrons from NADH or FADH2 to flavoproteins (FMN)
H+ pumped into periplasm
Electrons transported
To Iron-Sulfur proteins from NADH
To CoQ from FADH2
Cytochromes transfer electrons
Final Electron Acceptor
O2 if Aerobic
Other inorganic molecule if Anaerobic

46. Proton Pump

47. ETC: NAD NADH + H+

48. ETC: Cytochromes

49. Cytochrome Oxidase (Cyt a + Cyt a3)

50. Oxidase Test
Oxidase (+) Pseudomonas
Oxidase (-) E.coli, Proteus

51. ATP Synthetase

52. Anaerobic Respiration
Use of another compound than O2 as final electron acceptor in the ETC
Examples
Nitrate ion NO3- [Pseudomonas, E coli, Bacillus]
NO2-
N2O N2
Sulfate ion SO4= to H2S
Methanogens
Carbonate ion CO3= to CH4

53. Nitrogen Reduction Test
Nitrate Reduction
+ with Zinc
Nitrate Reduction
Neg with zinc
Nitrate Reduction +

54. Other Catabolic Processes
Protein
Deamination
Decarboxylation
Dehydrogenation

55. Other Catabolic Processes
Lipid
Glycerol FA
Beta oxidation
Acetyl CoA

56. Biosynthesis Polysaccharide Lipid Nucleotides Keto acids AA
Glycerol from FA
Lipid
Glycerol + FA
Keto acids + NH2
Nucleotides
Nitrogen bases from Keto acids + NH2
5 C sugars from alternate CH20 Metabolism

57. Functions Polysaccharide Cell wall components LPS Lipid Cell wall
Plasma membranes
AA/Protein
Cell wall / membrane components
Enzymes
Toxins
Nucleotide
DNA
RNA
ATP
NAD
NADP

58. Protein Synthesis and Enzyme Regulation

59. Metabolism
Summary

60. Questions?