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1 COURSE: MIB 303 Microbial Physiology and Metabolism (3 Units- Compulsory) Course Duration: Three hours per week for 15 weeks (45 hours). Lecturer: Jimoh, S.O. B.Sc., M.Sc, Ph.D Microbiology (ABU, Zaria) Microbiology Unit, Department of Biological Sciences, College of Natural and Applied Sciences, Fountain University, Osogbo, Nigeria. Consultation Hour: Tuesday 11am-2.00pm COURSE DETAILS Dynamics of growth. Nutrition and energy metabolism of microorganisms. Effect of physical and chemical factors on growth. Biochemistry of various microbial processes such as transport, regulation and respiration. Biosynthesis of microbial products. Buffer preparation and standardization. Basic separation techniques in microbiology, dialysis, salting out, gel filtration, electrophoresis etc. Assay techniques for various metabolites including microbial enzymes, acids etc. Course Description The course focuses on organized chemical activities performed by a cell, which comprise of energy production and energy utilization. Course Justification The multiplicity of processes performed by all biological systems is traced (directly or indirectly) to certain chemical reactions. The geometric structure of the rigid peptidoglycan component of the cell wall is also determined by chemical reactions involved in its synthesis. Energy is required for synthesis of enzymes, nucleic acids, polysaccharides and other chemical components; repair damaged cells, growth and multiplication. Thus, to support such extensive activities, vast amounts of energy must be provided through microbial metabolism. Methods of Grading S/N Style of grading Percentage Score 1 Attendance, class work and assignment 10 2 Test 20 3 Examination 70 Total 100 Course Delivery Strategies LECTURE CONTENT Weeks 1 and 2: Dynamics of growth (Microbial growth in closed and open systems). Objective: At the end of the lectures, the students shall be able to assay for microbial growth in batch culture and under constant environmental conditions maintained through continual provision of nutrients and removal of wastes (continuous culture).

2 Description First, second and third week (Week 1) Cultivation and growth pattern in batch culture First, second and third hours (Week 2) Continuous culture system using chemostat and turbidiostat. 1. Write short note on Unbalanced growth. 2. (a) A continuous culture system in a chemostat is controlled by dilution rate, explain. (b) Differentiate between chemostat and turbidostat. 3. Explain in details, the cultivation and growth pattern of microorganisms in batch culture. 4. What are the effects of dilution rate on microbial growth? 5. Mention the merits of continuous culture system. 6. Explain briefly, the increase in cellular constituents. 7. What are the parameters to consider when analyzing growth of a microbial culture? 8. What are the factors responsible for variation in lag phase of a closed system? Week 3: Nutritional requirement of microorganisms Objective: At the end of the lectures, the students shall be able to categorize microorganisms based on their sources of carbon, energy and electron. First hour (Week 3) Macronutrients and trace elements Second and third hours (Week 3) Metabolic diversity in microorganisms and growth factors 1. Microorganisms require nutrients for synthesis of new cellular components and energy regulation, Elaborate. 2. Discuss the metabolic diversity in microorganisms. 3. Write shorts on the major classes of growth factors. 4. Biosynthesis and energy production during microbial growth depends on nutritional factors, Discuss. Week 4: Mechanism of microbial control Objective: At the end of the lectures, the students shall be able to describe mechanism of action of chemical and physical agent for prevention or inhibition of microbial growth. First and second hours (Week 4) Effect of physical agents on microbial growth

3 Second hour (Week 4) Effect of chemical agents on microbial growth Third hour (Week 4) Conditions influencing the effectiveness of antimicrobial agents 1. Write short notes on the following; (i) Sterilization (ii) Disinfection (iii) Chemotherapy 2. Explain the mechanism of action of the following. (i) Cationic detergent (ii) Halogens (iii) Ethylene oxide gas (iv) Phenolics and alcohols (v) Moist heat 3. What are the characteristics of an ideal antimicrobial agent? Week 5: Metabolism of microbes Objective: At the end of the lectures, the students shall be able to describe control mechanisms that modulate the enzymatic composition of the cell. First hour (Week 5) Anabolic and Catabolic reactions in microbes Second and third hour (Week 5) Metabolic catalysts and enzymatic activities (effects of substrate concentration, ph, temperature, enzyme concentration on enzymatic activity First, second and third hour (Week 5) Regulation of enzyme activity i.e. feedback inhibition, precursor activation and energy -link control 1. Describe the following with the aid of appropriate illustration (i) Effect of substrate concentration on enzyme activity. (ii) Effect of temperature on enzyme activity 2. Differentiate between Feed-back inhibition, precursor activation and Energy- link control. 3. Write short note on energy- conserving reactions. 4. List the steps involved in synthesis of complex organic molecules. 5. Explain briefly the following; (i) Enzyme specificity (ii) Enzyme sensitivity 6. Write short notes on the following

4 (i) NAD + and NADP + (ii) FAD and FMN 7. Explain briefly the competitive and non-competitive inhibition. 8. Describe the factors that affect the rate of an enzyme catalyzed reaction. Week 6: Regulation of enzyme synthesis Objective: At the end of the lectures, the students shall understand the mechanism of enzyme synthesis and regulation. First, second and third hours (Week 6) Induction and repression of enzyme synthesis, end product repression and catabolite repression 1. Explain briefly, the induction and repression of enzyme synthesis. 2. Describe the mechanism involved in end product repression. 3. In a medium containing both glucose and lactose, which substrate is E coli likely to consumed first and why. 4. Describe the general properties of enzymes. 5. Explain the following; (i) Cofactor (ii) Coenzyme (iii)apoenzyme (iv) Prosthetic group Weeks 7, 8 and 9: Catabolic processes and metabolic role of ATP Objective: At the end of the lectures, the students shall be able to describe energy production by anaerobic and aerobic processes; and the flow of metabolites through these pathways under a high degree of regulation and control. First hour (Week 7) Substrate- level phosphorylation Second hour (Week 7) Oxidative phosphorylation Third hour (Week 7) Photophosphorylation First hour (Week 8) Glycolysis Second hour (Week 8) Entner-Doudoroff pathway Third hour (Week 8) Respiratory electron transport system

5 First hour (Week 9) Fermentation Second hour (Week 9) The tricaboxylic acid cycle Third hour (Week 9) Respiratory electron transport system 1. What is oxidative phosphorylation and where does it occur in the respiratory chain? 2. Briefly explain how glycolysis fits into the metabolism of glucose in aerobic cells. 3. Compare the disposition of electrons (or hydrogen atoms) obtained from oxidation from the oxidation ofglyceraldehyde-3-phosphate in aerobic and anaerobic cells. 4. Enumerate the three reactions in the glycolytic pathway that are not freely reversible by the same specific enzymes. 5. Describe the various ways in which the pentose phosphate cycle is useful to a cell. 6. Explain why fermentation is a less efficient process for obtaining energy than aerobic respiration. 7. Why is TCA cycle called an amphibolic cycle? 8. Describe the Embden- Meyerhof pathway with the aid of appropriate illustrations. 9. Discuss the anaerobic fates of pyruvic acid showing the reaction mechanism. Week 10: Continuous assessment Weeks 11 and 12: Biosynthetic Processes Objective: At the end of the lectures, the students shall be able to describe the biosynthetic processes of macromolecules. First hour (Week 11) Protein synthesis Second and third hours (Week 11) Synthesis of Carbohydrate First hour (Week 12) Lipid synthesis Second and third hours (Week 12) Nucleic acid synthesis 1. Outline the major steps involved in Protein synthesis. 2. What is the biological importance of monosaccharides? 3. Draw the structure of α-maltose and L- fructose. 4. Describe the major functions of lipids. 5. List the three major types of lipids in membrane.

6 6. Define peptide bond and differentiate between dipeptide, oligopeptide and polypeptide. 7. Describe the primary and secondary structures of proteins. 8. Describe the structure of RNA. 9. Discuss the DNA structure and its functions. 10. List the differences between RNA and DNA molecules. Weeks 13 and 14: Basic separation techniques in microbiology; Buffer preparation and standardization. Objective: At the end of the lectures, the students shall understand the significance of buffer in solution in biological system and the principal buffer systems in the body fluid; principles and applications of different chromatographic techniques and electrophoresis. First, second and third hours (Week 13) Principles of chromatographic techniques (gel filtration, absorption chromatography, partition chromatography and ion- exchange chromatography); principle and application of electrophoresis First, second and third hours (Week 14) Buffer preparation and standardization; Assay techniques for various metabolites including microbial enzymes, acids 1. Describe the preparation of 5dm 3 of a 0.3 mol/ dm 3 acetate buffer; ph 4.47, starting from a 2 mol/ dm 3 solution of acetic acid and a 2.5 mol/ dm 3 solution of sodium acetate. 2. Define a buffer. 3. What are the factors that determine the effectiveness of a buffer? 4. Explain the principles of chromatography 5. Compare and contrast thin layer chromatography and affinity chromatography 6. Define and state the basic laws of Spectrophotometry. 7. Describe the principles and application of centrifugation and electrophoresis. Week 15: Tutorials and Revision class Reading list / References

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