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Reading Guide for Week 4 – Bio260 Colleen Sheridan Stage 02 – Colonization and Infection Unfortunately, some bacteria managed to make it into our host’s body before we were able to take steps to reduce disease transmission. In this unit you will be trying to figure out how bacteria adhere to the host’s cells and grow to colonize the host. In last week’s reading guide you learned how our normal microbiota help to keep pathogenic microbes from adhering and growing. But, sometimes, pathogenic microbes make it past this line of defense and adhere to our cells. So we also learned how certain prokaryotic cellular structures were used to adhere to host cells to begin colonization. Then you learned of the incredible variety of different environmental conditions that different bacteria could grow (colonize) in. Now that our bacteria have attached and their environmental conditions are right, let’s go inside the bacteria to find out how they grow and get a sense of the immense diversity of prokaryotic metabolism. In this week’s reading you will learn about: 1. Transport of molecules into and out of the cell. 2. How chemoorganoheterotrophic bacteria utilize organic compounds that are transported into the cell to harvest energy and produce precursor metabolites (through catabolism) that are then used by the cell to synthesize subunits which are used to build macromolecules which are used to build cell structures (through anabolism) all driven by the activity of enzymes. 3. A brief look at the diversity of chemoorganotrophic prokaryotes. Chapter 3: Microscopy and Cell Structure (section 3.5, review from Week 1 reading) 1. Understand the different ways to transport material across a cytoplasmic membrane. Understand the different ways bacteria move material across a membrane such as facilitated diffusion and active transport mechanisms (transport systems that use proton motive force, transport systems that use ATP, group translocation, and efflux pumps). 2. What are the functions of the bacterial cytoplasmic membrane? Chapter 6: Metabolism: Fueling Cell Growth (whole chapter) 1. Define metabolism and describe the difference between anabolism and catabolism. Be able to draw and explain Figure 6.1. 2. Review the components of metabolic pathways, like enzymes, ATP, and electron carriers. 3. Understand a key concept to harvesting energy: the roles of the chemical energy source and the terminal electron acceptor and the oxidation-reduction reactions and electron carriers that transfer the electrons between them. What is reducing power? 4. What are precursor metabolites and where do they come from? No need to memorize the names of the precursor metabolites, but you should understand that the central metabolic pathways produce them using catabolic processes and that they go on to synthesize the subunits of the 4 main categories of biological macromolecules using anabolic processes: Subunits (made from precursor metabolites) amino acids nucleotides monosaccharides glycerol fatty acids Macromolecules (made from subunits) proteins nucleic acids carbohydrates lipids 5. Understand the following terms and processes at the level of detail found in the “Overview of Catabolism” section found on p.132-134: glycolysis, pentose phosphate pathway, Tricarboxylic acid cycle, respiration (anaerobic and aerobic respiration), and fermentation, and how the following processes play a role in those catabolic processes: oxidation/reduction reactions, electron transport chain, oxidative phosphorylation, and substrate level phosphorylation. 6. Define enzymes and their characteristics. What biological macromolecule is an enzyme made out of? 7. What are cofactors? What are coenzymes? 8. Understand the factors that influence enzyme activity: temperature, pH, substrate concentration, and inhibitors. 9. Understand competitive and noncompetitive inhibition. Know an example of each. 10. Know that cytochromes are oxidases and are components of the electron transport chain. Why do the oxidase test? 11. What is the difference between aerobic respiration, anaerobic respiration, and fermentation? How do they differ in the amount of energy (ATP) produced? What is the final electron acceptor in each pathway? 12. What are the uses of the proton motive force in prokaryotes? 13. What is the final end product from fermentation of glucose by E. coli? By Clostridium? By lactic acid bacteria? By Saccharomyces? How do the fermentation end products help us to explain our results on the EMB agar plate in Lab Ex. 7? 14. What type of energy harvesting pathway(s) might be used by chemoorganoheterotrophic - obligate aerobes? - obligate anaerobes? - facultative anaerobes? 14. Know that lipids, amino acids, and nucleotides are synthesized from precursor metabolites. Why do fastidious bacteria require growth factors? Chapter 11: The Diversity of Prokaryotic Organisms (11.1, 11.5-11.6, 11.8) No need to memorize the following, but become familiar enough with these so that, should an in-class exercise on prokaryotic diversity occur, you will be able to finish the project in a timely manner! Metabolic Diversity 1. Where can you find anaerobic environments in/on the human body? 2. Understand that some anaerobic chemoorganotrophs harvest energy through fermentation. Be able to briefly describe the genera: Clostridium and Propionibacterium, and the lactic acid bacteria (particularly Streptococcus) and give an example of each if one is provided in the book. 3. If you want to, skim through the rest of the sections not required and get an idea of the incredible metabolic diversity that prokaryotes have. 4. What is an aerobic chemoorganotroph? What is an obligate aerobe? Be able to briefly describe the genera: Micrococcus, Mycobacterium, and Pseudomonas and give an example of each if one is provided in the book. 5. What is a facultative anaerobe? Be able to briefly describe the genus Corynebacterium and the Family Enterbacteriaceae and give an example of each if one is provided in the book. What is an “enteric”? What are “coliforms”? Ecological Diversity 6. What soil living bacterial genera form endospores? Do you know if any of these cause human disease? 7. Describe the skin environment. Be able to briefly describe the genus Staphylococcus and give an example. 8. Describe where you would find mucous membranes in the human body. Be able to briefly describe the genera: Bacteroides, Bifidobacterium, Campylobacter, Helicobacter, Haemophilus, Neisseria, Mycoplasma, Treponema, Borrelia and Chlamydia and give an example of each if one is provided in the book.