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Glycolysis : Energy Generation Without an Oxygen Requirement

Glycolysis : Energy Generation Without an Oxygen Requirement. Glucose Biofuel Prominence : Low-reactive ring-form minimizes protein glycosylation. Glycolysis : A Three Step Process . Glucose trapping and destabilization (priming) Three carbon unit generation (cleaving)

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Glycolysis : Energy Generation Without an Oxygen Requirement

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  1. Glycolysis: Energy Generation Without an Oxygen Requirement Glucose Biofuel Prominence: Low-reactive ring-form minimizes protein glycosylation

  2. Glycolysis: A Three Step Process • Glucose trapping and destabilization (priming) • Three carbon unit generation (cleaving) • Energy generation

  3. Induced Fit in Hexokinase Glucose induces a large enzyme conformational change Substrate-induced cleft closing prevents ATP hydrolysis Kinases require a divalent metal ion What function does Mg+2 play in hexokinase?

  4. Hexokinase Closed Around Substrates What mechanisms of catalysis are operative?

  5. Hexokinase Reaction Mechanism What is the Nu:, electrophile, and leaving group in this reaction?

  6. PhosphoglucoseIsomerase: Aldose to Ketose Conversion

  7. PhosphoglucoseIsomerase (PGI) G6P Conversion via Acid-Base Catalysis PGI Reaction Mechanism

  8. PhosphoglucoseIsomerase (PGI) G6P Conversion via Acid-Base Catalysis Base catalyzed bond formation

  9. PhosphoglucoseIsomerase (PGI) G6P Conversion via Acid-Base Catalysis Acid catalyzed ketal formation

  10. PhosphoglucoseIsomerase (PGI) G6P Conversion via Acid-Base Catalysis Base catalyzes ring closure H+

  11. Phosphofructokinase: Trapping the Fructose Isomer What is the mechanism for this reaction?

  12. Glycolysis Stage I: Glucose Trapping and Destabilization (priming)

  13. Six Carbon Sugar Cleaved to Two Three Carbon Units What is the bond to be cleaved? Which alcohol becomes an aldehyde?

  14. Haworth and Fischer Projections Equivalency The functional group that is down in a Haworth projection is positioned how in a Fischer structure?

  15. Aldolase Reaction Mechanism Aldolase Fructose-1,6-bisphosphate binds to the aldolase enzyme for covalent catalysis

  16. Aldolase Reaction Mechanism What is lost when the Schiff base forms?

  17. Aldolase Reaction Mechanism Aldolase Rxn Mechanism Compare and contrast a Schiff base with a carbonyl group.

  18. Aldolase Reaction Mechanism What is the process for Schiff base to carbonyl conversion?

  19. Aldolase Reaction Mechanism H2O Aldolase cleaves FBP into GAP and DHAP

  20. Triose Phosphate Isomerase (TIM) Reversible and driven towards GAP due to product depletion Which previous glycolytic step is similar to TIM?

  21. Triose Phosphate Isomerase Reaction Mechanism Glycolysis: Step #5Triose Phosphate Isomerase TIM- or α,β-barrel with 8 parallel β-strands surrounded by 8 α-helices. DHAP conversion to GAP necessary to proceed through glycolysis

  22. Stoichiometry: Stages 1-2 of Glycolysis Two ATPs are initially invested. One glucose is metabolized into two GAP molecules.

  23. Road Map for Energy Harvest (Stage 3)

  24. Glyceraldehyde-3-Phosphate Dehydrogenase: Covalent Catalysis

  25. Glyceraldehyde-3-Phosphate Dehydrogenase: a 2 Step Process What amino acid will serve as a nucleophile to form a thioester?

  26. Glyceraldehyde-3-Phosphate Dehydrogenase: Reaction Mechanism

  27. Glyceraldehyde-3-Phosphate Dehydrogenase: Catalysis Energetics Hypothetical reaction with no coupling Actual coupled reaction

  28. Phosphoglycerate Kinase What is the Nu:, electrophile and leaving group for this reaction? (hint: consider hexokinase in reverse)

  29. Glycolysis: the Three Final Steps

  30. Pyruvate Kinase What is the Nu:, electrophile and leaving group for this reaction? (hint: consider phosphoglycerate kinase)

  31. Glycolysis Energetic ∆G°ʹ ∆G Enzyme (kcal/mol) (kcal/mol) 1near equilibrium means that ∆G is about zero What is the relationship between ∆G and ∆G°ʹ? When can ∆G and ∆G°ʹ diverge?

  32. Regulating Glycolysis: A Pictorial Analogue • Water represents • metabolite flux • Water amount in • flask represents • intermediate • abundance • Flasks connections • are enzymes • Vertical drop represents • decrease in free energy • ΔG° = height difference between flask bottoms • ΔG = height difference between water levels

  33. Alternative Fates for Pyruvate

  34. Anaerobic Recycling of NADH for Glycolysis

  35. Microbial Recycling of NADH for Glycolysis

  36. Pathogenic Obligate Anaerobes

  37. Starting and Ending Point for Fermentation Not Always Same

  38. Chapter 16, Problems 1-12

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