Introductory Microbiology Chap. 5 Chapter Outlines/Notes Introduction What are you and microbes and all living things made of, i.e., what forms the structures in a living cell, and what performs living functions? What do living cells have to do to obtain the things they need to form the above structures and perform the above functions? What molecules allow organisms to acquire the things they need for life processes at the time they need them? How does the (above answer) function to do that? So cells need and . What determines what molecule source or energy source a specific species can use? Both are central to METABOLISM. Define. Slide 1, 2 All living cells’ energy ‘currency’: Note: Cells use only two kinds of energy: 1) light energy: trapped and used by plants and some bacteria for photosynthesis and 2) chemical energy: the energy held in the bonds of various chemicals. Cells do not use thermal or electrical energy because they don't have thermal or electrical converters. Thermal potential (that is, temperature) affects the rate of chemical reactions, but does not provide any energy. What about the electrical signals of nervous impulses? They use energy in the form of ATP to generate electric potentials in the membrane of nerve cells and fibers. Those electrical signals are not ‘used’ by the cell to perform other work. 1 Chemical reactions are central to LIFE: Catabolic and Anabolic Reaction Pathways Slides 3 & 4 Chapter 5 Microbial Metabolism p. 113 Most processes are the same/similar in all cells, but not all. Microbes can do things we cannot do! Crazy things, like eat petroleum or radioactive materials and things that are waste products to us. Online Site http://www.aw-bc.com/microplace/ Pretest #1-10, 14-16 I. Catabolic and Anabolic Reactions p. 114 Slides 3 & 4 A. METABOLISM : Two categories of Chemical Processes/Reactions Catabolic and Anabolic Reactions Metabolism and Metabolic Pathways Slide 4 2 Chapter 5 Microbial Metabolism I. II. III. IV. V. VI. VII. VIII. IX. X. XI. Catabolic and Anabolic Reactions Enzymes Energy Production Carbohydrate Metabolism Lipid and Protein Catabolism Biochemical Tests and Bacterial Identification (not on exam) Photosynthesis A Summary of Energy Production Mechanisms Metabolic Diversity among Organisms Metabolic Pathways of Energy Use The Integration of Metabolism Online Site http://www.aw-bc.com/microplace/ Pretest #1-10, 14-16 I. Catabolic and Anabolic Reactions A. SLIDES 3 & 4 METABOLISM : Two categories of Chemical Processes/Reactions 1. Catabolic and Anabolic Reactions 2. Metabolism and Metabolic Pathways 3 B. Molecules and Energy- intertwined…. See Text Fig. 5.1 Role of ATP See similar Fig. 2.18 on p. 49 Textbook Online Site http://www.aw-bc.com/microplace/ Metabolism Overview Animation with Quizzes 4 II. Enzymes: How they function and what can affect them Enzymes increase the rate of chemical reactions by decreasing the activation energy required for that specific reaction. A. Collision Theory SLIDE 5 B. Enzymes and Chemical Reactions Catalysts Substrate Decrease activation energy C. Enzyme Specificity and Efficiency SLIDE 6 Specificity Turnover number D. Naming Enzymes See Table 5.1 p. 116 Enzyme Classification Based on Type of Chemical Reaction Catalyzed E. Enzyme Components Slide 7 Fig. 5.3 p. 116 Apoenzyme (protein portion) Cofactor (nonprotein portion) Haloenzyme F. The Mechanism of Enzymatic Action: The sequence of events in enzyme action on the reactant(s), the enzyme’s substrate(s) Lock and Key Model SLIDE 8 Fig. 5.4 p. 118 5 G. Factors Influencing Enzyme Activity End of Chap. ‘Study Questions’ Review #2 a-c SLIDE 9, 10 Fig. 5.6 p. 119 1. Example of Competitive Inhibition and its use in Medicine: Sulfa Drugs SLIDE 11 Fig. 5.7 p 120 End of Chap. ‘Study Question’ Critical Thinking #2 SLIDES 12-14 Structure: Sulfa drugs are chemically similar (chemical analogues) to the chemical PABA (para-aminobenzoic acid) which is required by microbes to produce folic acid. Folic acid is a vitamin that functions as an enzyme cofactor in the synthesis of nucleotide nitrogen bases. p. 120 Normal functioning when no sulfa drugs are present: PABA is used by bacteria to produce folic acid. (Table 5.2 p. 117) When sulfa drugs are present: Since sulfa drugs are chemically similar to (analogues of) PABA, they may “trick” the enzyme into using the sulfa drug to produce folic acid instead of PABA. In effect, the sulfa drug competes with the PABA. When the enzyme is ‘tricked’, the folic acid in the bacterial cell is defective and will not produce nitrogen bases. It there is a shortage of nitrogen bases, DNA cannot be replicated, and therefore, growth stops (no binary fission). The bacterial cell has been inhibited. Some disadvantages of sulfa drugs: 1) Only bacteriostatic, not bactericidal 2) Resistance 3) Hypersensitivity 4) Crystallization in kidney of patient 6 2. Noncompetitive Inhibition See Fig. 5.7c p. 120 Allosteric site H. Feedback Inhibition Example: Production of the amino acid isoleucine See Fig. 5.8 p. 121 I. Ribozymes What are they and what chemical reactions do they catalyze? See Learning Objectives p. 115 and Check Your Understanding p. 121 7 III. Energy Production A. Oxidation-Reduction Reactions SLIDE 15 & 16 SLIDES 20-23 Fig. 5.9 p. 122 Come back to this in a minute… End of Chapter ‘Study Questions’ Multiple Choice #1 Removal and Gain of B. The Generation of ATP: Phosphorylation of ADP to ATP SLIDE 17 1. Substrate-level phosphorylation SLIDE 18 2. Oxidative phosphorylation SLIDE 24 3. Photophosphorylation SLIDE 19 End of Chapter ‘Study Questions’ Review #5 See Learning Objectives p. 121 and Check Your Understanding pp. 122, 123 Textbook Online Site http://www.aw-bc.com/microplace/ Energy Production o Animations with Quizzes: Oxidation-Reduction Reactions o Interactive Tutorials: Oxidation-Reduction 8 C. Metabolic Pathways of Energy Production In the cell, there are many series of enzymatically catalyzed chemical reactions that store energy and release energy from organic molecules- carbohydrates, proteins, and lipids. Catabolic reactions with these molecules release energy for ATP production and anabolic reactions use the energy in those ATPs primarily to synthesize large forms of these molecules. IV. Carbohydrate Metabolism A. Glycolysis SLIDE 25 SLIDE 26-32 Fig. 5.12, Text p. 126 Biochemical pathway (10 reactions) in which ONE molecule of GLUCOSE is OXIDIZED to form 2 molecules of PYRUVIC ACID. NAD (nicotinamide adenine dinucleotide) is reduced: Important electron carrier ATP is used and formed B. Alternatives to Glycolysis Text discussion p. 125, 127 SLIDE 30 9 NEXT: Either Cellular Respiration or Fermentation C. Cellular Respiration 1. Aerobic respiration Define respiration: p. 127 a. Glycolysis b. Intermediate Step after Glycolysis SLIDE 31 Fig. 5.13 p. 128 c. Krebs cycle SLIDES 32, 33 Fig. 5.13 d. Electron transport chain/ Chemiosmosis/ADP Phosphorylation 2. Electron transport chain: a series of electron carriers are oxidized (lose electrons) & other electron carriers are reduced (gained electrons) as electrons are passed from one electron carrier to another. NADH and FADH2 will donate their electrons to electron carriers located in the prokaryotic plasma membrane. Eukaryotes- occur in the mitochondria. Oxygen is the final electron acceptor in aerobic respiration. Before continuing, turn the page and draw the electron transport chain. Then come back! Info: Some electron carriers pick up one electron, others pick up more than one electron. Some electron carriers carry hydrogen atoms (1e-, 1p+); others ONLY CARRY ELECTRONS (THE PROTON IS SEPARATED FROM THE ELECTRON IN THE H ATOMS). In the electron transport system, FADH2 donates its electrons after NADH There also is anaerobic respiration where the final electron acceptor is NOT oxygen. Less common, doesn’t produce as much ATP. 10 Electron Transport Chain 11 Chemiosmosis Fig. 5.15 p. 130, Fig. 5.16 p. 131 1) Occurs simultaneously with the electron transport chain (they are coupled) to transfer the energy to form ATP from ADP and phosphate (ADP phosphorylation). 2) At certain points along the electron transport chain, the hydrogen atom is ‘split’; the electron and the proton are separated. Remember, some electron carriers carry hydrogen atoms (1e-, 1p+). So what happens to the protons in the H atoms? 3) The protons are pumped out of the cell (through the plasma membrane) & the electron is then passed to other electron carriers. 4) This creates a situation where there are more protons on the external side of the plasma membrane than are on the internal side of the plasma membrane; in other words, a gradient is formed. 5) This gradient creates a force. 6). Chemiosmosis is the process of creating a proton gradient (by movement of protons across the plasma membrane) and the subsequent movement of the protons back into the cell through specific channels. Surrounding those channels is ATP synthase. ATP Generation: ADP Phosphorylation 1) Bonding of a phosphate group to ADP to form ATP. 2) The energy required to bond the phosphate to ADP (and that is then stored in the resulting bond) is provided by the movement of protons back into the cell during chemiosmosis. 3) When the protons rush back into the cell (due to the gradient), energy is released. This may cause a CONFORMATIONAL (shape) change in the enzyme ATP synthase. ATP synthase that then catalyzes the reaction: SLIDES 34-37 Fig. 5.15 p. 130 and Fig. 5.16 p. 131 Overview: Electron Transport Chain & Chemiosmotic Generation of ATP SLIDE 38 Comparing Eukaryotes and Prokaryotes: Where in the cell? SLIDE 39 Carbohydrate Catabolism – Number of ATPs generated from aerobic respiration through Substrate-Level Phosphorylation Only SLIDE 40 Carbohydrate Catabolism – Number of ATPs generated from aerobic respiration through Substrate-Level and Oxidative Phosphorylation 12 Overall Summary Reaction of Aerobic Respiration: 13 Anaerobic Respiration. SLIDE 41, 42 What takes the place of oxygen? The amount of energy generated varies depending on the electron acceptor. EXAMPLES: a. Sulfate. In marine sediments this leads to large amounts of sulfate reduction - Sulfate SO42- is converted (reduced) to hydrogen sulfide H2S - which some may be familiar with as the rotten egg smell and black material that can be found just a few centimeters below sediment surfaces. b. Nitrate NO3-. which is converted (reduced) to nitrite NO21-, nitrous oxide N2O, or nitrogen gas N2 in the process. c. Metal ions. For example: Fe+2, Mn+2 d. Carbonate, CO32-. is converted (reduced) to methane, CH4. This is called methanogenesis Very little energy is obtained from methanogenesis and vast amounts of substrate need to be turned over to make a living. What are these organisms? Primarily live in conditions. 14 D. Fermentation Definition SLIDE 43 Steps of Fermentation SLIDE 44 Fig. 5.18 p. 134 Two Primary Categories of Fermentation SLIDE 45 Fig. 5.18 p. 134 SLIDE 46 Fig. 5.19 p. 136 Examples of Types & Importance of Fermentation SLIDE 47 Table 5.4 p. 137 SLIDE 48 Table 5.4 p. 137 Table 5.5 p. 137 Aerobic Respiration, Anaerobic Respiration, and Fermentation Compared Fig. 5.11 p. 125 Foundation Figure An Overview of Respiration and Fermentation End of Chapter ‘Study Questions’ Review #4 a and b Multiple Choice #7-10 Textbook Online Site http://www.aw-bc.com/microplace/ Microflix 3-D Animation Cellular Respiration Carbohydrate Catabolism o Animations with Quizzes Glycolysis #3-5 Electron Transport Chain Overview #1-5 Electron Transport Chain The Process #1 & #4 o Interactive Tutorials: Fermentation See Learning Objectives p. 124 and Check Your Understanding pp. 127, 132 15 V. Lipid and Protein Catabolism Learning Objective p. 136 SLIDE 32 Summary of the interrelationships of carbohydrate, lipid and protein catabolism Fig. 5.21 p. 138 16 VI. Biochemical Tests and Bacterial Identification (not on exam) Lab- Your Unknown VII. Photosynthesis p. 140 SLIDES 50-53 Textbook Online Site http://www.aw-bc.com/microplace/ Photosynthesis o Animations with Quizzes Overview Comparing Prokaryotes and Eukaryotes Learning Objective 5-20 & 5-21 and Check Your Understanding p. 140 VIII. A Summary of Energy Production Mechanisms p. 141 See Requirements for ATP (energy sources, electron carriers, final electron acceptors) Fig. 5.27 p. 143 Recap what is required for ATP Production End of Chapter ‘Study Questions; Multiple Choice #3 Learning Objective and Check Your Understanding p. 141 17 IX. Metabolic Diversity among Organisms p. 142 Metabolic Classification: Nutritional Patterns SLIDE 54 Fig. 5.28 p. 143 (old Slides 55 & 56 combined) Nutritional Classifications of all organisms: Energy Source (see below) Nutritional Classifications of all organisms: Carbon Source (see below) Put Energy and Chemical Requirements together to classify organisms by their nutritional needs SLIDE 55 Summary Table of Fig. 5.28 p. 143 End of Chapter ‘Study Questions’ Review #7 Learning Objective p. 142 X. Metabolic Pathways of Energy Use p. 146 Now you have ATP, what are you going to do with it? Describe the major types of anabolism and their relationship to catabolism. 18 XI. The Integration of Metabolism p. 147 SLIDE 56, 57 Fig. 5.33 p. 149 Amphibolic pathways: Metabolic pathways that have both catabolic and anabolic functions Learning Objectives: pp. 146 and 147 and Check Your Understanding 5-24 p. 147 19 SLIDE 58 Metabolism The Big Picture End of Chapter ‘Study Questions’ Review #1h Textbook Online Site http://www.aw-bc.com/microplace/ Concept Map (left side column): Metabolism Step 2 o MP3 Tutor Session o Integration of Metabolism o Animations with Quizzes Metabolism: The Big Picture Step 3: Test Yourself o Multiple Choice #1, 9, 11, 12, 13, 15, 18, 19, 20 o Fill in the Blank #1-4, 6, 9, 10 o True/False #3, 4, 8, 9 o Foundation Figure Quiz #1-4 o Microbe Review #2 & 3 o Post-Test #1-3, 5, 6, 8-14, 18, 20 20