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Exploring Microbial Metabolism and Enzyme Functions in Energy Generation Pathways

Discover the biochemical diversity of microbial metabolism, the relationships within metabolic pathways, and the various forms of energy generation processes. Learn about enzymes' characteristics, functions, and factors affecting their activity. Uncover the nuances of glycolysis, fermentation, aerobic and anaerobic respiration, and other metabolic pathways in microbes. Dive into the role of enzymes in redox reactions, electron transport chains, and metabolic diversity.

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Exploring Microbial Metabolism and Enzyme Functions in Energy Generation Pathways

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  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 Polymerase Others: Lyases, Hydrolases, Isomerases, Transferases

  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 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 Compared

  20. 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

  32. Acetyl CoA Formation

  33. Acetyl CoA Final Structure

  34. Krebs CycleTCA/Citric Acid Eukaryotic Mitochondria Or Prokaryotic Cytoplasm

  35. Oxidation in Krebs Cycle

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

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

  38. Redox Reactions

  39. Oxidation

  40. Reduction

  41. Catabolism + Anabolism

  42. Eukaryotic ETC

  43. 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)

  44. 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

  45. Proton Pump

  46. ETC: NAD NADH + H+

  47. ETC: Cytochromes

  48. Cytochrome Oxidase (Cyt a + Cyt a3)

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

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