Micrococcus luteus on blood agar - PowerPoint PPT Presentation

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Micrococcus luteus on blood agar

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  1. Micrococcus luteus on blood agar

  2. A microbiologists view of the periodic table Group 11 12 13 4 1 2 14 15 16 17 3 7 8 10 6 18 9 5 Period Key: 1 Essential for all microorganisms Essential cations and anions for most microorganisms Trace metals, some essential for some microororganisms 2 Used for special functions Unessential, but metabolized 3 Unessential, not metabolized 4 5 6

  3. Colony morphology , A form of multicellularity?

  4. Isolated colonies at end of streak Confluent growth at beginning of streak Streaking for singles. Looking for single colony forming units

  5. Enzymes lower activation energy Activation energy— no enzyme Substrates (A  B) Activation energy with enzyme Free energy ∆G0= Gf0(C  D) Gf0(A  B) Products (C  D) Progress of the reaction

  6. Enzymes are recycled Glyceraldehyde-3-P Dihydroxyacetone-P Substrate Fructose 1,6-bisphosphate Products Active site Enzyme–substrate complex Free aldolase Free aldolase

  7. Enzymes are specific for their substrates 3 dimensional structure determined by folding is dependent on side chain interactions determined by charge and hydrophobicity.

  8. Leo the lion goes Gerrrrrr ReDox - gaining electrons = reduction losing electrons = oxidation Electron acceptor Electron donor Electron-donating half reaction Electron-accepting half reaction Formation of water Net reaction

  9. E0(V) Redox couple -0.60 -0.50 -0.40 Some ReDox potentials of ETC -0.30 (1) -0.20 -0.10 0.0 +0.10 (2) +0.20 +0.30 +0.40 +0.50 +0.60 +0.70 (3) +0.80 +0.90 (1) H2fumarate2succinate2 ∆G0  = –86kJ ∆G0  = –163kJ (2) H2 NO3 NO2+ H2O ∆G0  = –237kJ (3) H2 O2H2O

  10. Fig. 5-10-1 E0(V) Redox couple -0.60 -0.50 -0.40 -0.30 (1) -0.20 -0.10 0.0 +0.10 (1) H2fumarate2succinate2 ∆G0 = –86 kJ

  11. NADH  H Fig. 5-11 Reduced Oxidized NAD Nicotinamide Ribose Ribose Adenine Phosphate added in NADP

  12. Reaction 1. Enzyme I reacts with electron donor and oxidized form of coenzyme, NAD+. Reaction 2. Enzyme II reacts with electron acceptor and reduced form of coenzyme, NADH. NADH binding site Active site NAD+ binding site Active site Fig. 5-12 Enzyme II Enzyme I NADH Electron acceptor Electron donor NAD+ Enzyme substrate complex NADH Electron acceptor reduced Electron donor oxidized NAD+

  13. Bond energies of some important compounds Ester bond Anhydride bonds Ester bond Anhydride bond Adenosine triphosphate (ATP) Glucose 6-phosphate Phosphoenolpyruvate Anhydride bond Thioester bond Acetyl Coenzyme A Acetyl phosphate Acetyl-CoA

  14. Anhydride bonds Ester bond Fig. 5-13-1 Anhydride bond Adenosine triphosphate (ATP) Phosphoenolpyruvate Anhydride bond Thioester bond Coenzyme A Acetyl Acetyl phosphate Acetyl-CoA

  15. Intermediates in the biochemical pathway Energy-rich intermediates Using SLP to drivethermodynamically unfavorable reactions Substrate-level phosphorylation Energized membrane Less energized membrane Oxidative phosphorylation

  16. STAGE I: PREPARATORY REACTIONS Isomerase Phosphofructokinase Hexokinase Glucose Fructose-6- Fructose-1,6- Glucose-6- You must use energy to free energy Aldolase STAGE II: MAKING ATP AND PYRUVATE Glyceraldehyde-3- 2 Glyceraldehyde-3-P dehydrogenase 2 Electrons 2 NAD+ 1,3-Bisphosphoglycerate 2 NADH 2 To Stage III Phosphoglycerokinase 2 3-Phosphoglycerate 2 2-Phosphoglycerate Enolase 2 Phosphoenolpyruvate STAGE III: MAKING FERMENTATION PRODUCTS Pyruvate kinase Pyruvate 2 NADH Lactate dehydrogenase Pyruvate decarboxylase Pyruvate:Formate lyase NAD+ To Stage II Acetate formate Formate hydrogenlyase CO2 Acetaldehyde Lactate H2 CO2 NADH Alcohol dehydrogenase To Stage II NAD+ Ethanol

  17. Fig. 5-15-1 STAGE I: PREPARATORY REACTIONS Isomerase Phosphofructokinase Hexokinase Glucose Fructose-1,6- Glucose-6- Fructose-6-

  18. Investment and return on investment Aldolase STAGE II: MAKING ATP AND PYRUVATE Glyceraldehyde-3- 2 Glyceraldehyde-3-P dehydrogenase 2 Electrons 2 NAD+ 2 1,3-Bisphosphoglycerate 2 NADH To Stage III Phosphoglycerokinase 3-Phosphoglycerate 2 2 2-Phosphoglycerate Enolase 2 Phosphoenolpyruvate

  19. Fig. 5-15-3 STAGE III: MAKING FERMENTATION PRODUCTS Pyruvate kinase Pyruvate 2 NADH Lactate dehydrogenase Pyruvate decarboxylase Pyruvate:Formate lyase NAD+ To Stage II Acetate formate Formate hydrogenlyase CO2 Lactate Acetaldehyde H2 CO2 NADH Alcohol dehydrogenase To Stage II NAD+ Ethanol

  20. R-Cysteine Cysteine-R Iron-sulfur clusters : a motif for electron transfer R-Cysteine Cysteine-R R Cysteine R Cysteine R Cysteine Cysteine R

  21. E0(V) Complex I Fig. 5-20 –0.22 0.0 Complex II Fumarate Succinate CYTOPLASM 0.1 Complex III Complex IV 0.36 0.39 ENVIRONMENT E0(V)

  22. chemiosmosis F1/Fo ATP synthase and the proton gradient F1 In b2 Membrane Fo C12 Out

  23. Pyruvate (three carbons) The Balance sheet: The bottom line Energetics Balance Sheet for Aerobic Respiration Key C2 2 Pyruvate  4 ATP  2 NADH (1) Glycolysis: Glucose  2NAD 2 ATP Acetyl-CoA C4  4 ADP C5 to CAC to Complex I C6 (a) Substrate-level phosphorylation 2 ADP  Pi 2 ATP 8 ATP Oxalacetate2 (b) Oxidative phosphorylation Citrate3 2 NADH 6 ATP Aconitate3 3 CO2 4 NADH FADH GTP  (2) CAC: Pyruvate 4 NAD GDP  FAD  Malate2 Isocitrate3 to Complex I to Complex II (a) Substrate-level phosphorylation Fumarate2 1 GDP  Pi 1 GTP 1 ATP  1 GDP 1 GTP  1 ADP 15 ATP ( 2) (b) Oxidative phosphorylation 4 NADH 1 FADH 12 ATP 2 ATP Succinate2 –Ketoglutarate2 Succinyl-CoA 38 ATP per glucose (3) Sum: Glycolysis plus CAC

  24. Energetics Balance Sheet for Aerobic Respiration Key  4 ATP 2 NADH 2 Pyruvate (1) Glycolysis: Glucose  2NAD 2 ATP Fig. 5-22b  4 ADP C2 to CAC to Complex I C4 (a) Substrate-level phosphorylation C5 2 ATP 2 ADP  Pi 8 ATP C6 (b) Oxidative phosphorylation 6 ATP 2 NADH 3 CO2 4 NADH FADH (2) CAC: Pyruvate 4 NAD GDP  FAD GTP    to Complex II to Complex I (a) Substrate-level phosphorylation 1 GTP 1 GDP  Pi 1 GTP  1 ADP 1 ATP  1 GDP 15 ATP ( 2) (b) Oxidative phosphorylation 4 NADH 1 FADH 12 ATP 2 ATP 38 ATP per glucose (3) Sum: Glycolysis plus CAC

  25. Fermentation Organic compound CO2 Carbon flow in respirations Electron transport/ Proton motive force Biosynthesis O2 Fig. 5-23 Organic e– acceptors Aerobic respiration Electron acceptors S0 NO3– SO42 Anaerobic respiration Chemoorganotrophy Inorganic compound CO2 Electron transport/ Proton motive force Carbon flow Electron acceptors Biosynthesis O2 S0 NO3– SO42 Chemolithotrophy Photoheterotrophy Photoautotrophy Light Electron transport Organic compound CO2 Carbon flow Carbon flow Proton motive force Biosynthesis Biosynthesis Phototrophy

  26. Organic compound Fermentation CO2 Carbon flow in respirations Electron transport/ Proton motive force Fig. 5-23ab Biosynthesis O2 Organic e– acceptors Aerobic respiration Electron acceptors NO3– S0 SO42 Anaerobic respiration Chemoorganotrophy Inorganic compound CO2 Electron transport/ Proton motive force Carbon flow Electron acceptors Biosynthesis O2 S0 NO3– SO42 Chemolithotrophy

  27. Fig. 5-23c Photoautotrophy Photoheterotrophy Light Electron transport Organic compound CO2 Carbon flow Carbon flow Proton motive force Biosynthesis Biosynthesis Phototrophy

  28. Glutamate family Proline Glutamine Arginine -Ketoglutarate Fig. 5-25 Citric acid cycle Aspartate family Asparagine Lysine Methionine Threonine Isoleuine Oxalacerate Alanine family Valine Leucine Pyruvate Glycolysis Serine family Glycine Cysteine 3-Phosphoglycerate Phospho- enolpyruvate Aromatic family Phenylalanine Tyrosine Tryptophan Chorismate Erythrose-4-P

  29. Fig. 5-26 Glutamate -Ketoglutarate  NH3 Glutamate dehydrogenase Glutamine Glutamate NH3 Glutamine synthetase Glutamate Oxalacetate -Ketoglutarate  Aspartate Transaminase 2 Glutamate Glutamine  -Ketoglutarate Glutamate synthase

  30. CO2 Amino group of aspartate Glycine Formyl group (from folic acid) Formyl group (from folic acid) Fig. 5-27 Amide nitrogen of glutamine Ribose-5-P Inosinic acid Aspartic acid NH3 CO2 Orotic acid Uridylate

  31. Acetyl-ACP Malonyl-ACP Fig. 5-28 Acetoacetyl-CoA Palmitate (16 C) 4 C 6 C 14 C 8 C 12 C 10 C

  32. Starting substrate The allosteric enzyme Control of pathways: feedback inhibition (noncompetitive inhibition) Enzyme A Intermediate I Enzyme B Intermediate II Feedback inhibition Enzyme C Intermediate III Enzyme D End product

  33. Enzyme Active site Allosteric site Fig. 5-30 End product (allosteric effector) Substrate INHIBITION: Substrate cannot bind; enzyme reaction inhibited ACTIVITY: Enzyme reaction proceeds

  34. Erythrose 4-phosphate Phosphoenol pyruvate Initial substrates  3 1 2 Fig. 5-31 DAHP synthases (isoenzymes 1, 2, 3) DAHP Chorismate Tyrosine Tryptophan Phenylalanine

  35. Glutamine synthetase, a paradigm of allosteric control 100 Enzyme activity Glutamine Glutamine concentration Relative GS activity Glutamine 50 GS–AMP6 GS–AMP12 GS 0 3 6 9 12 0 AMP groups added AMP

  36. The makings of a microbe

  37. also

  38. Cofactors galore: Take your vitamins!

  39. Tab. 5-4

  40. Thinking thermo!