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Chapter 5: Microbial Metabolism What is Metabolism? Metabolism = all chemical reactions that occur in a cell/organism; catabolism + anabolism. Catabolism = breakdown of food molecules to produce energy and molecular subunits (example: amino acids from proteins) Anabolism = building of macromolecules that the organism needs (proteins from amino acid subunits or DNA from nucleotides) Metabolic (also called biochemical) pathways http://www.expasy.ch/cgi-bin/show_thumbnails.pl e1 A e2 B e3 C D Feedback inhibition = shutting off of biochemical pathway; product in pathway binds to e1 which changes its shape and stops its work Enzymes Type of molecule – usually proteins; a few RNA molecules called ribozymes too. Words ending with –ase are enzymes Shape determines function Active site E = enzyme, S = substrate, P = product E+S ES E+P Enzyme catalyzed reactions Enzyme-substrate complexes What affects enzyme activity? Temperature pH Concentration of substrate Inhibitors Ex. Folic acid pathway Sulfanilamide competes with PABA; no folic acid made Ex. Fluoride in drinking water and toothpaste inhibit cavity-causing bacteria Denaturation of proteins How temperature, pH, and substrate concentration affect enzyme activity -- How inhibitors work -- Energy molecules in cells ATP ADP + Pi NADH NAD+ + H+ + 2eFADH2 FAD + 2H+ + 2e- Catabolism of Glucose Aerobic respiration – efficient energy production (1 glucose converted to 38 ATP) 2. Anaerobic respiration – less efficient (less than 38 ATP); no oxygen 3. Fermentation – least efficient (2 ATP produced); usually without oxygen 1. Aerobic Respiration Process: Start molecule End molecule Energy molecules produced Waste Glycolysis Glucose Pyruvate 2 NADH 2 ATP -- Oxidation of pyruvate Pyruvate Acetyl-CoA 2 NADH 2 CO2 Krebs cycle (citric acid cycle) Acetyl-CoA -- 6 NADH 2 ATP 2 FADH2 4 CO2 Electron transport chain/chemiosmosis NADH FADH2 NAD+ FAD ATP Water Anaerobic respiration Uses parts of all of the aerobic processes (but not all) Final electron acceptor not oxygen Possible e- acceptors: Nitrogen compounds NO3- NO2, N2O, N2 Sulfur compounds SO4-2 H2S Carbon compounds CO3-2 CH4 Fermentation Only produces 2 ATP molecules from 1 glucose Occurs without oxygen (O2) Detour from glycolysis to a short fermentation pathway Produces by-products as NAD+ and FAD are regenerated By-products produced include acids, alcohols, gases, etc. Practical example: Fact: Clostridium perfringens causes gas gangrene Problem: How to cure gas gangrene… Patients affected: Anyone with an anaerobic site of tissue damage; diabetics particularly prone to this disease (Diabetics have poor circulation, nerve damage, lessened pain sensations, lower ability to heal) Diagnosis: Odors, specimen collection and identification of bacteria, X-ray for pockets of gas in tissues Treatment: Essential -- Remove dead tissue (sometimes even amputation of affected limb); Possible -- expose tissues to oxygen through a hyperbaric chamber and/or antibiotic regimen This photomicrograph reveals Clostridium perfringens grown in Schaedler’s broth using Gram-stain. (Photo by CDC/Don Stalons, 1974.) Wagner Grade 5 - Unsalvagable Gangrene Gangrene or necrosis to the extent that the foot is beyond salvage and will require a major limb- or life-sparing amputation. http://www.squidoo.com/diabeticfoot Why do microbes have the ability to switch back and forth between aerobic respiration and an anaerobic process? Process Advantages Disadvantages Aerobic respiration Makes many ATPs from 1 glucose Requires oxygen; produces oxygen free radicals Anaerobic processes (fermentation or anaerobic respiration) Survival without oxygen Makes less ATP from glucose; slower growth; d etrimental waste products What about other food sources besides glucose? What about anabolic reactions? The End