2-co so thu nghiem sinh hoa-Microbial Metabolism.ppt

2-co so thu nghiem sinh hoa-Microbial Metabolism.ppt

1. Microbial Metabolism
Biochemical diversity

2. Metabolism Relationships

3. Metabolism Pathways

4. Metabolism
Define
Requirements
Energy
Enzymes
Rate
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
Kinetic
Potential
Use
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
Protein
Ribozyme [ribosome]
Characteristic functions
Active site
Specific
Modified Forms
Inactive
Active
Coenzyme/Cofactor
-ase
Others: Lyases, Hydrolases,
Isomerases, Transferases
Polymerase

9. Enzyme Characteristics

10. Enzyme Structure
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
@ active site
Similar shape
Binding
Irreversible
Reversible
Noncompetitive
@ allosteric site
Change shape
Binding
Irreversible
Reversible

13. Noncompetitive Enzyme Inhibition

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

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 Compared
Pathway EM PPP ED
Location
Substrate
Cytoplasm
Glucose
Cytoplasm
G6P
Cytoplasm
Glucose
Steps 2 parts
6C 3C
Different
sugars (4-7C)
Glyceraldehyd
e
Products 2
Pyruvate
2 ATP
2 NADH
2 H+
Pyruvate &
Fructose
6 CO2
12 NADPH
12 H+
2 Pyruvate
1 ATP
1 NADPH
1 NADH
2 H+

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

21. Anaerobic Processes
Lactic Acid
Lactobacillus
Mixed Acid
Enterobacteriaceae
Butanediol
Klebsiella
Enterobacter
Butyric Acid
Clostridia
Butanol-Acetone
Clostridia
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
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
Methanogens

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

54. Other Catabolic Processes
Protein
Deamination
Decarboxylation
Dehydrogenation

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

56. Biosynthesis
Polysaccharide
Keto acids
AA
Glycerol from FA
Lipid
Glycerol + FA
AA
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