Friday, March 2, 2012

Course Syllabus: TU M.Sc Microbiology First year:Biochemistry and Instrumentation


Biochemistry and Instrumentation
Course Title: Biochemistry and Instrumentation Full Marks: 100
Course No: MB 513                                                        Pass Marks: 40
Nature of Course: Theory Year: I

Objectives

Upon completion of the course, students will have
  • Advanced knowledge on general and microbial biochemistry.
  • Advanced knowledge on principles and procedures of various biochemical techniques and instrumentation required for conducting analysis and research.
Overview of Principles of Biochemistry, Bio-molecules 2 hrs

Carbohydrates
Classification, Structures and Biological functions of Carbohydrates and Glycoproteins 6 hrs

Amino Acids and Proteins
Classification, Structure and Biological function: Amino acids, Protein, Peptides and Polypeptides. Methods for characterization and Purification of proteins 3 hrs

Lipids and Fatty Acids
Biological role of lipids. General properties, distribution, classification and nomenclature of lipids, Structure and properties of neutral fats and phospholipids, Glycolipids, steroid, Structural components of lipids: Hydrophobic components, Fatty acids with even and odd number of carbon atoms saturated and unsaturated fatty acids, Fatty alcohols, glycerol, diols, inositol, carbohydrate components, amino alcohol/sphingozine/aminoacids, phosphates, sulfates. Neutral lipids, Ethers, Steroid derivatives, Fatty acids and their role in lipid metabolism 6 hrs

Enzymes: Kinetics and Regulation
Nomenclature and principle of enzyme classification, Henri equation, Michaelis-Menten equation; kinetics of enzymatic reactions involving two substrates, Factors affecting enzymatic activity and kinetics, Mechanisms of enzyme catalysis; Structure and mechanisms of lysozyme; serine proteases and glutathione reductase, immobilized enzymes. Allosteric regulation, rate limiting enzymes, Isozymes and their role, Enzymes of microbial origin and their applications 8 hrs

Biological Membrane
Biological functions, Structure and properties of membrane lipids, Formation of artificial membrane and their applications. Dynamic properties of membrane lipids. Classification, characteristics and distribution of membrane proteins, extraction and isolation of membrane proteins, Movements of main proteins and lipids, Asymmetry of membrane, Factors influencing on membrane fluidity 3 hrs

Vitamins and Coenzymes
Role of vitamins, metals and other cofactors in enzyme functions. Types, properties and classification of Vitamins: Water- soluble vitamins and their coenzymes, Lipid-soluble vitamins, Iron containing coenzymes and metal cofactors. 8 hrs


Nucleic Acids
Structure of Nucleic acids, Purines and pyrimidines bases, Carbohydrate components, Mononucleotides, Nucleoside: mono, di-and tri-phosphates, DNA and RNA, their localization in cell 2 hrs

Metabolism of carbohydrates, Proteins, and Lipids
Basic mechanisms of metabolism and energy conversion, Metabolism and metabolic pathways. Linear, cyclic and spiral metabolic pathways, ATP as a cellular energy currency, NAD+ and NADP+/NADH and NADPH (Oxidizing and reducing agents) 2 hrs
Metabolism of Carbohydrates
Intercoversion of carbohydrates. Coenzymatic functions of nucleotides (e.g. UTP, UDP). Anaerobic and aerobic degradation of carbohydrates. Different types of fermentation. Glycolysis. Oxidative phosphorylation in the level of substrate, Glyconeogenesis Oxidative decarboxylation of pyruvic acid, Pyruvate dehydrogenase complex, Cycle of di-and tri-carbonic acids and their biological role, Terminal processes of oxidation. Electron transport system, Energetic significance of cascade process of electron transport from substrate to oxygen. Oxidative phosphorylation of respiratory chain, Concept of conjugated oxidation and phosphorylation in respiratory chain, Transmembrane potential of hydrogen ions, Energetic effect of glycolysis and respiration. Gluconeogenesis: substrates of gluconeogenesis (lactate, pyruvate and other C3 compounds, acetate, glyoxylate). 6 hrs

Metabolism of Amino acids and Proteins
Enzymatic hydrolysis of proteins, proteolytic enzymes. Degradation of amino acids, Transamination and its mechanism, Various types of deamination, Urea cycle, Link between urea cycles and Kreb's cycle. Metabolism of Ammonia. Deamination and carboxylation of aminoacids, Amines and their physiological role, Proteineous nitrogeneous substances: biosynthesis, degradation and biological functions 6 hrs

Metabolism of Lipids
Enzymatic degradation of triglycerols and absorption by cell membrane. Oxidation of fatty acids, ß-oxidation mechanism in prokaryotes and eukaryotes, Oxidation of unsaturated fatty acids, Biosynthesis of fatty acids, Biosynthesis of neutral fats, phospholipids, and cholesterol 8 hrs

Nucleic Acids Metabolism
Mechanism of biosynthesis of nucleosides and degradation of nucleotides 4 hrs

United System of Process of Metabolism
Integrated system of metabolism of proteins, carbohydrates and lipids 2 hrs

Hormones
General Characteristics, Classification, Mechanism of action 4 hrs
Instrumentation

Biochemical Techniques
General principle and approaches to biochemical investigations, In vivo/ in vitro studies, physiological solutions. Buffer solutions, Tissue homogenization and separation 3 hrs
Cell Disruption
Methods of disruption of tissues and cells, Protein purification: Background factors, protein extraction, preliminary fractionation procedures, chromatographic and electrophoretic techniques, monitoring the purification process 4 hrs

Working Principle, Instrumentation and Application of -Phase contrast and Electron Microscopy, Gel-Doc system 3 hrs

Principles, types and uses of -Centrifugation techniques, Gel filtration, Electrophoretic techniques: Starch gel, Agarose gel, Cellulose Acetate, Polyacrylamide gel Electrophoresis, Isoelectric focusing, Two- dimensional Electrophoresis, Blotting technique 10 hrs

Principles, types and uses of Chromatographic Techniques

Basic concepts and instrumentation- Ion exchange chromatography, Affinity Chromatography, Partition chromatography, Paper and Thin layer chromatography, Gel Permeation chromatography, Gas Chromatography, High performance Liquid Chromatography (HPLC) 10 hrs

Instrumentation, Working Principle and Application of- Atomic absorption spectroscopy, Atomic emission Spectroscopy- Flame photometry, Plasma emission Sprectroscopy, Fluorimetry and spectrofluorimetry 10 hrs

Principle, Instrumentation and Application of- Ultra-violet and visible spectrometry, Infra-red spectroscopy, Molecular luminescence spectroscopy, Nuclear Magnetic resonance spectroscopy, Mass Spectroscopy 10 hrs

Textbooks

  1. Nelson DL and Cox MM. Lehninger Principles of Biochemistry, 5th Edition, Freeman (2004).
  2. Wilson K and Walker J (Eds). Principles and Techniques of Biochemistry and Molecular Biology, 6th Edition, Cambridge University Press (2005).
  3. Voet D and Voet J. Biochemistry. 3rd Edition, Wiley International Edition (2004).
  4. Stryer L. Biochemistry, 4th Edition, W.H. Freeman Company, New York (1995).
  5. Plummer DT. An Introduction to Practical Biochemistry, 3rd Edition, Tata McGraw Hill (1988).
  6. Skoog DA, Holler FJ and Nieman TA. Principles of Instrumental Analysis, 5th Edition, Thomson Books/Cole (2005).

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