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
- Nelson DL and Cox MM. Lehninger Principles of Biochemistry, 5th Edition, Freeman (2004).
- Wilson K and Walker J (Eds). Principles and Techniques of Biochemistry and Molecular Biology, 6th Edition, Cambridge University Press (2005).
- Voet D and Voet J. Biochemistry. 3rd Edition, Wiley International Edition (2004).
- Stryer L. Biochemistry, 4th Edition, W.H. Freeman Company, New York (1995).
- Plummer DT. An Introduction to Practical Biochemistry, 3rd Edition, Tata McGraw Hill (1988).
- Skoog DA, Holler FJ and Nieman TA. Principles of Instrumental Analysis, 5th Edition, Thomson Books/Cole (2005).
- Mendham J, Denny RC, Barnes JD and Thomas M. Vogel's Text Book of Quantitative Chemical Analysis, 6th Edition, Pearson Education (2008).
More Syllabus of M.Sc First Year Microbiology (Tribhuvan University)
- Syllabus of M.Sc First year Immunology (TU)
- Syllabus of M.Sc First year Biochemistry-Instrumentation
- Syllabus of Microbial Biotechnology and Pharmaceutical Microbiology
- Syllabus of Microbial Structure, Physiology and Genetics
- Syllabus of Epidemiology, Research Methods and Biostatistics
- Syllabus of Practical Course on (MB 513+MB 514)
- Syllabus of Practical Course on (MB511+MB 512)
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