Metabolism Energy Pathways Biosynthesis. Catabolism Anabolism Enzymes
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1 Topics Microbial Metabolism Metabolism Energy Pathways Biosynthesis 2 Metabolism Catabolism Catabolism Anabolism Enzymes Breakdown of complex organic molecules in order to extract energy and dform simpler end products. Enzymes are involved. 3 4 Metabolism Model Enzymes Function Structure Enzyme-substrate interaction Cofactors Action Regulation 5 6 1
2 Enzyme Structure Conjugated Enzymes Simple enzyme - primarily protein Conjugated enzyme - protein and nonprotein Three-dimensional features: Specificity - lock-and-key Active site or catalytic site Conjugated enzymes contain a metallic cofactor, coenzyme, or both in order for it to function as a catalyst. 7 8 Active Site Enzyme-Substrate Interaction Substrates specifically bind to the active sites on the enzyme: - lock-and-key style -Induced fit Once the reaction is complete, the product is released and the enzyme reused. Specific active sites are folded regions of the protein molecule and contain specific amino acids in its microenvironment Lock-and-Key Model Coenzymes Function as transient carriers Alter a substrate by removing a chemical group from it and adding it to another. Ex. NAD, FAD and CoA Specificity of enzyme-substrate reactions and induced fit
3 Coenzyme Activity Enzyme Action Carrier function of coenzymes A coenzyme transfers chemical groups from one substrate to Exoenzymes Endoenzymes Constitutive Induction or repression Types of reactions another Enzyme Location Constitutive and Regulated Enzymes Constitutive enzymes are present in constant amounts. Exoenzymes are inactive while inside the cell, but upon release from the cell they become active. Endoenzymes remain in the cell and are always active. 15 Regulated enzymes are either induced or repressed. 16 Types of Reactions Synthesis and Hydrolysis Condensation Hd Hydrolysis Transfer reactions 17 Condensation reactions are associated with anabolic reactions, and hydrolysis reactions are associated with catabolic reactions. 18 3
4 Transfer Reactions Sample Enzymes Transfer of electrons from one substrate to another. Ex: Oxidoreductase - oxidation-reduction reactions. Transfer of functional groups from one molecule to another. Ex: Aminotransferases - transfer of amino group. Examples of enzymes, their substrates, and their reactions Regulation Patterns of Metabolism Metabolic pathways Direct control Genetic control Metabolic pathways follow stepwise patterns. These are regulated by enzymes that catalyze these reactions Enzyme Control Mechanisms Genetic Control Repression - end products stop the expression of genes that encode for proteins (enzymes) which are responsible for metabolic reactions. Induction - substrate initiates and enhances the expression of genes for proteins (enzymes) that drive metabolic reactions. Competitive inhibition and noncompetitive inhibition are forms of direct control (regulation) of the enzyme action
5 Repression Enzyme Characteristics Repression as a type of genetic control of enzyme synthesis Bioenergetics Energy Machinery of the Cell Cell energetics - Exergonic reactions - Endergonic reactions Redox reaction Electron carriers Adenosine Triphosphate (ATP) 27 The general scheme associated with metabolism of organic molecules, the redox reaction, and the capture of energy in the form of ATP. 28 Redox Reaction Electron Carriers Oxidation - removal or loss of electrons Reduction - addition or gain of electrons These are coupled reactions Biological redox reactions involve transfer of electrons and protons (hydrogens) = dehydrogenation Dehydrogenases - catalyze these reactions Electron carriers - transfer electrons (and protons) from donor to acceptor molecules. Coenzymes: Ex: Nicotinamide adenine dinucleotide (NAD) Respiratory chain (ETC) carriers: Ex: Cytochromes (protein+porphyrin)
6 Adenosine Triphosphate Energy Capture Temporary energy repository ( cellular battery ) Breaking of pyrophosphates bonds will release free energy for cellular work. Three part molecule: Nitrogen base - Adenine Pentose sugar - Ribose) Chain of three phosphate groups The phosphate groups capture the energy derived from metabolism as pyrophosphate bonds within the ATP molecule. ATP and its partner compounds ADP and AMP Phosphorylation Substrate-level Phosphorylation ATP can be used to phosphorylate an organic molecule such as glucose during catabolism. ATP can be synthesized by substrate-level phosphorylation. Phosphorylation - catalyzed by phosphorylases (e.g. hexokinase) 33 A phosphate group from an intermediate is transferred to ADP to regenerate ATP. 34 Catabolic Pathways Glucose Metabolism Embden-Meyerhoff-Parnas (EMP) Pathway or Glycolysis Kreb s s or Tricarboxylic Acid (TCA) Cycle Overview of the location, flow, endproducts of cellular Electron Transport or Respiratory Chain (aerobic) Alternate pathways respiration. Fermentation Glucose is catabolized to harness energy
7 Cellular Respiration Glycolysis Glycolysis Kreb s Cycle Electron Transport Chain 37 Glucose (6-carbon sugar) splits into two pyruvates (3-carbon molecules). Glucose is oxidized and coenzyme NAD is reduced to NADH. Energy investment phase: - Phosphorylation of intermediates using 2 ATP molecules Energy yielding phase: - Substrate-level-phosphorylation of ADP to produce 4 ATPs. 38 Glycolytic Steps Kreb s Cycle Each pyruvic acid is processed to enter the Kreb s Cycle as Acetyl CoA. CO 2 is generated -decarboxylation reactions. Coenzymes NAD and FAD are reduced to NADH and FADH 2 Net yield of two ATPs per molecule of glucose Steps in Kreb s Cycle Electron Transport Chain NADH and FADH 2 from glycolysis and Kreb s Cycle donate electrons to the electron carriers (ETC). Membrane bound carriers transfer electrons by redox reactions. Oxygen (final electron acceptor) completes the terminal step
8 Electron Transport Chain Location of ETC Eukaryotes - Inner Mitochondrial Membrane Prokaryotes- Cytoplasmic Membrane The Electron Transport Chain and Chemiosmosis driven by the Proton Motive Force ATP Yield Anaerobic Respiration Glycolysis - 2 Kreb s Cycle - 2 ETC- 34 Total Yield: 38 Similar to aerobic respiration, except nitrate or nitrite is the final electron acceptor NADH yield - FADH 2 yield- 2 in Glycolysis 8 in Kreb s Cycle 2 in Kreb s Cycle Fermentation Fermentation Glycolysis only NADH from glycolysis is used to reduce the glucose Organic compounds as the final electron acceptors (not O 2 ) Low ATP yields per glucose molecule compared to cellular respiration Chemistry of fermentation: Production of ethyl alcohol or lactic acid and release of CO
9 Types of Fermenters Facultative anaerobes Fermentation in the absence of oxygen Respiration in the presence of oxygen Ex. Escherichia coli Strict fermenters No respiration Ex. yeast Fermentation Products Alcoholic fermentation Acidic fermentation Mixed acid fermentation Mixed Acid Fermentation Biosynthesis Mixed acid fermentation and related products synthesized from pyruvate Amphibolic Gluconeogenesis Beta oxidation Amination Transamination Deamination Macromolecules Amphibolic Synthesis Amphibolic Synthesis Integration of the catabolic and anabolic pathways (Coupled Reactions) Intermediates serve multiple purposes Intermediates serve as precursors to synthesize amino acids, carbohydrates and lipids
10 Gluconeogenesis Beta Oxidation Pyruvate (intermediate) is converted back to glucose Metabolism of fats into acetyl, which can then enter the Kreb s cycle as acetyl CoA. Occurs when the glucose supply is low Amino Acid Synthesis Macromolecules Cellular building blocks: Monosaccharides Amino acids Fatty acids Nitrogen bases Vitamins
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