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Topics • Metabolism = Catabolism + –Metabolism Anabolism –Energy –Pathways • Enzymes

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Topics • Metabolism = Catabolism + –Metabolism Anabolism –Energy –Pathways • Enzymes Chapter 8 Metabolism Topics • Metabolism = Catabolism + –Metabolism Anabolism –Energy –Pathways • Enzymes Catabolism Anabolism • Enzymes are involved in the • Enzymes are involved in the use breakdown of complex organic of energy from catabolism in molecules in order to extract order to synthesize energy and form simpler end macromolecules and cell products structures from precursors (simpler products) Catabolism and Anabolism - what does it take to make a living bacterium? Enzymes Function • Catalysts for chemical reactions – EX. - RedOx reactions • Lower the energy of activation 1 Enzyme Structure Enzyme Structure Enzyme-substrate interactions Two components: 1) Apoenzyme – protein component • Substrates specifically bind to the active sites on the enzyme 2) Cofactors - Coenzymes – Organic molecules (vitamins) • Once the reaction is complete, the product is released and the enzyme - Inorganic elements – Metal ions reused - over and over again! 3) Active site or Catalytic site – interacts with substrate Role of Enzymes in Disease E + Substrate ES Complex E + Products Secretion of Exoenzymes Avoid Host Defenses Virulence factors or Toxins Multiply in Tissues Examples: Streptococcus pyogenes (Strept Throat) Produces Streptokinase - digests clots Pseudomonas aeruginosa (many diseases) Produces Elastase and Collagenase - digest elastin and collagen - Disease Clostridium perfringens (gas gangrene) Lecithinase C - destroys tissue in wounds 2 Regulation Factors that Affect Enzyme • Metabolic pathways • Direct control - Quickest way to regulate • Temperature – Competitive - bind active site • pH – Noncompetitive - DO NOT bind active site • Osmotic pressure • Genetic control - Slower because 'switch' is mediated by accumulation or production of product Denaturation!!!! – Induction – Repression Competitive vs. Noncompetitive Inhibition Conserved Patterns in Metabolism * Linear Cyclic Branched -Divergent -Convergent Genetic control - more long term effects Lactose Operon • If E. coli into medium containing • Repression LACTOSE – Excess of end product binds to the • E. coli will synthesize the enzyme genetic component and turns if 'OFF' LACTASE • Induction • This enzyme will allow the organism to – Presence of substrate (LACTOSE) binds break down LACTOSE into GLUCOSE and to the genetic component and turns it GALACTOSE 'ON' • If LACTOSE is gone, LACTASE synthesis – Make enzymes necessary for action stops 3 Genetic Repression Energy • Cell energetics – Exergonic - RXN produces energy – Endergonic - RXN uses energy • Redox reaction • Electron carrier • Adenosine Triphosphate (ATP) Simplified Model of Energy Machine of Cell Redox reaction • Reduction and oxidation reaction • Electron carriers transfer electrons and hydrogens – Electron donor (REDUCED) – Electron acceptor (OXIDIZED) • Energy is also transferred and captured by the phosphate in form of ATP Adenosine Triphosphate Electron carriers –shuttle (ATP) electrons and hydrogens • Temporary energy repository • Coenzymes • Breaking of phosphates bonds will – Nicotinamide adenine dinucleotide release energy (NAD NADH) • Three part molecule – Flavin adenin dinucleotide (FADH ) – Nitrogen base • Respiratory chain carriers – 5-carbon sugar (ribose ) – Cytochromes (protein) – Chain of phosphates 4 How ATP is made? Remember, there is a determined number of ATP The molecules in the cell!! phosphates capture the • ATP is made either by SUBSTRATE energy and LEVEL PHOSPHORYLATION - donor becomes substrate part of the • or by OXIDATIVE ATP PHOSPHORYLATION - molecule respiration/electron transport system Glucose to Glucose-6-phosphate Phosphate comes from ATP Pathways • Catabolism A summary of –Glycolysis the metabolism –Tricarboxylic acid cycle (TCA) (catabolism ) of –Respiratory chain - Electron Transport glucose and •Aerobic the synthesis •Anaerobic of energy –Fermentation Anabolism - building blocks 5 Aerobic respiration Glycolysis • Oxidation of glucose • Glycolysis • Phosphorylation of some intermediates (Uses • Tricarboxylic acid (TCA) two ATPs) • Electron transport • Splits a 6 carbon sugar into two 3 carbon molecules (pyruvic acid) • Coenzyme NAD is reduced to NADH * Oxygen final electron acceptor • Substrate-level-phosphorylation (Four ATPs * ATP per glucose molecule = 38 are synthesized) ATP • NET YIELD = 2 ATP + 2 NADH + 2 pyruvic acid TCA cycle Summary of • Each pyruvic acid is processed to Acetyl- CoA (2C) that enters the TCA cycle (two Glycolysis complete cycles) • CO 2 is generated • Coenzymes NAD and FAD are reduced to NADH and FADH 2 • Net yield of 2 ATPs • Critical intermediates are synthesized 1 TCA cycle - remember - Electron transport this happens twice - once • NADH and FADH 2 donate electrons to the for each electron carriers pyruvic acid • NADH generates 3 ATP, FADH 2 = 2 ATP • Membrane bound carriers transfer Production of electrons (redox reactions) 8 NADH+ & 2 • The final electron acceptor completes the FADH 2 terminal step (ex. Oxygen) important - electron • Chemiosmosis transport • Proton motive force (PMF) system 6 The electron transport system and Electron transport chain oxidative phosphorylation • Mitochondria – eukaryotes • Cytoplasmic membrane – prokaryotes Chemiosmosis Final Step - - pump production hydrogen ions of ATP and out H2O Proton Motive Force - concentration gradient of differing charge ETS in a bacterial cell - Gram positive or negative? 7 Fermentation Anaerobic respiration • Glycolysis only • Incomplete oxidation of glucose in the absence of oxygen • Similar to aerobic respiration, • NADH from glycolysis is used to reduce the organic products except that oxygen • Organic compounds as the final electron containing ions such as acceptors nitrate or nitrite is the final • ATP yields are small ( 2 per glucose molecule), compared to respiration electron acceptor • Must metabolize large amounts of glucose to produce equivalent respiratory ATPs Types of fermenters Products of fermentation • Facultative anaerobes • Alcoholic fermentation – Fermentation in the absence of oxygen • Acidic fermentation – Respiration in the presence of oxygen – Ex. Escherichia coli • Mixed acid fermentation • Strict fermenters – No respiration – Ex. yeast Alcoholic and Acidic Fermentation Miscellaneous products of pyruvate - Mixed Acid Fermentation 8 Biosynthesis - Crossing pathways of Metabolism Examples of building Macromolecules - Catabolism and Anabolism - what does it amino acids take to make a living bacterium? 9.
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