NORTH MAHARASHTRA UNIVERSITY

JALGAON

NAAC ACCREDITTED

SYLLABUS FOR

Master of Science

In

Biochemistry
PART- I

(Semester I and II) w.e.f. 2007-2008

NORTH MAHARASHTRA UNIVERSITY, JALGAON M.Sc. Biochemistry Part-I

(W.e.f. 2007-08)

Course Structure at a Glance Semester I BC-101 : Enzymes and Enzyme technology BC-102 : Plant Biochemistry, Biostatistics & Bioinformatics BC-103 : Metabolism BC-104 : Microbial Physiology BC-105 : Lab. Course - I Semester II BC-201 : Human Physiology & Biochemistry of specialized tissues BC-202 : Analytical Biochemistry BC-203 : Immunology BC-204 : Lab. Course - II BC-205 : Lab. Course - III

Semester I and IV will have 04 theory courses and 1 practical course each whereas semester II and III will have 3 theory courses and 2 practical courses each. Practical examination will be held annually. Each course will have a weightage of 100 marks (20 marks internal and 80 university examination) Each theory course to be completed in 50 lectures of 60 min duration each

BC-101 : Enzymes and Enzyme technology

Enzyme classification, Characteristics, enzyme-substrate complex, concept of active centre, binding sites, stereospecificity ES complex formation, effect of temperature, pH and substrate concentration on reaction rate, activation energy, transition state theory. Enzyme acivity, International units, specific activity, turnover number. Enzyme kinetics: Michelis - Menten equation and its derivation, steady state enzyme kinetics, significance of Vmax and Km, Bisubstrate reaction , examples and reaction rate equations. Enzyme inhibition, types of inhibitors-competitive, non competitive and uncompetitive inhibition of enzymes, examples and experimental determination. Enzyme catalysis: factors affecting catalytic efficiency of enzymes- proximity and orientation effect, distortion or strain, acid base and nucleophilic catalysis. Isoenzymes and multiple forms of enzymes. Methods for studying fast reactions. Mechanism of catalysis of lysozyme, ribonuclease, chymotrypsin, and carboxypeptidase. Multienzyme complexes- pyruvate dehydrogenase and fatty acid synthetase, advantages of multienzyme complex. Enzyme regulation: general mechanism of enzyme regulation, feedback inhibition, feed forward stimulation, enzyme reparation , induction and degradation, enzyme regulation by cAMP, covalent modification and allosteric regulation of enzymes Kinetics and physiological significance of allosteric enzymes, symmetric and sequential model for action of allosteric enzymes Enzyme immobilization: various methods of immobilization, industrial application of immobilized enzymes. Biosensors: concept and application.

Recommended Books; 1. 2. 3. 4. 5. Fundamentals of Enzymology : N. C. Price and L. Stevens Enzymes : M. Dixon and E. C. Webb Biochemistry: L. Stryer Lehningers Principles of Biochemistry : D. L. Nelson and M. M. Cox Immobilised Biocatalyst : W. Hartneir ---------------------------------

BC 102 : Plant Biochemistry, Biostatistics and Bioinformatics

Section A : Plant Biochemistry Photosynthesis: Structure of chloroplast, photosynthetic pigments, excitation of chlorophyll molecule, photosystem I & II location, mechanism of quantum capture & energy transfer between photosystem, photolysis of water. Dark Reaction : C 3, C 4 and CAM metabolism, Photorespiration. Light activation of enzymes, regulation of photosynthesis & photorespiration. Bacterial photosynthesis. Nitrogen fixation : Symbiotic & non-symbiotic nitrogen fixation, nitrogenase & nitrate reductase, organization of nitrogen-fixation genes, nod genes, nif genes. Plant Hormones : Biochemical nature, mode of action and physiological effects/role of Auxins, Cytokinins, Gibberlins & Absisic acid. Secondary metabolites : Introduction, types, physiological functions & applications of alkaloids, flavanoids, terpenoids, tannins & lignins. Biofertilisers : Introduction, types, importance, mycorrhizal fungi. Section B : Biostatistics and Bio-informatics Basic terms, measures of central tendency and dispersion: Population, sample, variable, parameter, primary and secondary data, screening and representation of data. Frequency distribution, tabulation, bar diagram, histograms, pie diagrams, cumulative frequency curves,. Mean, median, mode, quartile and percentiles, measures of dispersion like range, variance, standard deviation, coefficient of variation. Probability and distribution : Definition of probability, addition and multiplication rules, binomial, Poisson and normal distribution. Correlation co-efficient, linear regression, regression co-efficient and coefficient of determination. Bioinformatics : History, web services as bioinformatics tools- alignment tools, the FASTA family, PatMatch, BLAST, ParAlign, ProteinEngine and Transeq, INTERPRO, CLUSTALW, PHYLIP, BioNumeric version 2.0, Biotechnix 3d.

Recommended Books : 1. 2. 3. 4. Plant biochemistry : Hans-Walter Heldt Handbook of Photosynthesis : Mohammed Pe Sarakle Introduction to Plant Biochemistry : T. W.Goodwin and E. L. Mercer Biochemistry of energy utilization in plants : D. T. Blackie

5. Plant Biochemistry : P. M. Dey and Harborne 6. Fertilizers : H. J. Tandon. 7. Computational Biochemistry : C. Stan Tsai 8. Fundamentals of Statistics : S. C. gupta 9. Biostatistics : Khan 10. Biostatistics : W. W. Daniel 11. Biostatistics : P. N. Arora & P. K. Malhan --------------------------------------

BC-103 : Metabolism
Laws of thermodynamics, entropy, enthalpy, Gibbs energy, free energy changes and redox potentials, phosphate group transfer potential. The mitochondrial respiratory chain , organization of intermediates, , ATP synthatase complex, theories of oxidative phosphorylation , uncouplers and inhibitors of oxidative phosphorylation, substrate level phosphorylation. Carbohydrate metabolism: Glycolysis and citric acid cycle, their regulation and role in energy generation, pentose phosphate pathway and its regulation, alternate pathways of carbohydrate metabolism. Gluconeogenesis, interconversion of sugars, biosynthesis of glycogen, and starch . Hormonal regulation of carbohydrate metabolism. Metabolism of Lipids: Fatty acid biosynthesis, fatty acid synthase complex, regulation of fatty acid biosynthesis. Desaturase and elongase and their roles. Biosynthesis of triacylglycerols, phospholipids, Formation and utilization of ketone bodies, metabolism of circulating lipids. Fatty acid oxidation. Amino acid metabolism: biosynthesis and degradation of amino acid and their regulation. Transamination and oxidative deamination. Urea cycle and its regulation. Inborn errors of amino acid metabolism. Nucleic acid metabolism ; Biosynthesis and catabolism of purines and pyrimidines and their regulation. Ribonucleotide reductase, biosynthesis of ribonucleotides and deoxyribonucleotides. Inhibitors of nucleic acid biosynthesis.

Recommended Books: 1. 2. 3. 4. Biochemistry: L. Stryer. Lehningers Principles of Biochemistry : D. L. Nelson and M. M. Cox. Harpers Biochemistry: R. K. Murray et al. (eds). Principles of Biochemistry : G. L. Zubay.

----------------------------

BC-104 : Microbial Physiology

Microbial systematics and taxonomy: New approaches to bacterial taxonomy , classification including ribotyping, ribosomal RNA sequencing, characteristics of primary domains , conventional and molecular taxonomy , the species concept, nomenclature & Bergeys manual. Methods in Microbiology: Pure culture techniques, Theory and practice of sterilization, principles of microbial nutrition ,construction of culture media , enrichment culture techniques for the isolation of chemoautotrophs and photosynthetic micro organisms. Microbial Growth: Definition of growth , mathematical expression of growth , growth curve, measurement of growth & growth yield , synchronous growth , continuos culture, growth as affected by environmental factors like temperature , acidity, alkalinity, water availability and oxygen, effect of toxic environments on microbial growth like heat, pasteurization, low temperature, sound , radiation , magnetic field and hydrostatic. Prokaryotic cells: Structure, function- cell walls of eubacteria (peptidoglycan) and related molecules , outer membrane of Gram negative bacteria, Flagella and motility, Cell inclusions like endospores, gas vesicles. Viruses : Discovery, classification and structure of viruses, bacteria , plant , animal and tumour viruses. Lysogeny, DNA viruses positive strand, negative strand and doubke stranded viruses, RNA viruses: adenoviruses, retroviruses , Viroids and Prions. Microbial Diseases : Disease reservoirs , epidemological terminologies, infectious disease transmission, respiratory infections caused by bacteria and viruses, tuberculosis , sexually transmitted diseases including AIDS. Diseases transmitted by: animals rabies, plague ; insect and ticks - rickettsias, lyme disease, malaria; food and water borne diseases. Pathogenic fungi. Emerging and recurrent infectitious diseases. Public health and water quality. Host- parasite relationships : Microbial interactions with higher organisms, normal microflora of the skin, oral cavity, gastrointestinal tract. Entry of pathogens into the host, Colonization and Growth. Types of toxins ( exo, endo, entero ), their structure and mode of action. Virulence and pathogenesis Chemotherapy/ Antibiotics : Antimicrobial agents , sulpha drugs , penicillins and cephalosporins. Broad spectrum antibiotics, mode of action. Resistance to antibiotics

Recommended Books : 1. General Microbiology, : Stainer, R.Y. , Ingraham, J.L., Wheelis , M.L. and P. R.Painter

2. Brock Biology of Microorganisms : Madigan, M.T.Marinko, J.M. and Parker, J. 3. Microbiology : Pelzar, M. J. Jr.,Chan , E. C. S. and Kreig, N. R. 4. General Microbiology : Robert. F . Boyd 5. .Microbial Physiology : G. Moat, John. W. Foster ----------------------------------------

BC-105 : Lab. Course I

1. Preparation of liquid and solid media for growth of microorganisms 2. Isolation and maintenance of microorganisms by plating, streaking and serial dilution methods 3. Biochemical characterization of selected microbes 4. Isolation of pure cultures from soil and water 5. Growth curve : measurement of bacterial population by turbidometric and serial dilution 6. Effect of temperature, pH, carbon and nitrogen sources on bacterial growth 7. Study of micro-organisms by Gram stain, acid fast stain and staining for spores 8. Demonstration of anti-bacterial activity & MIC of a commercial antibiotic 9. Isolation and culture of a common fungal strain 10. Demonstration of antifungal activity & MIC of commercial fungicide 11. Analysis of water for potability and determination of MPN. 12. Estimation of nitrogen by Kjeldal method 13. Spectral characteristics and estimation of chlorophyll 14. Determination of PS II activity 15. Bioefficacy of biofertilizers Recommended Books : 1. 2. 3. 4. 5. 6. 7. 8. 9. Laboratory manual of microbiology : J.G.Cappuccino and N Sherman Bacterological Techniques by F.J. Baker Introduction to Microbial Techniques : Gunasekaran Experiments in Microbiology, plant pathology, tissue culture and Mushroom cultivation : K.R. Aneja Laboratory manual in Biochemistry : J. Jayraman An introduction to Practical Biochemistry : D. Plummer Practical Biochemistry : K Wilson and Walker Biochemical Methods : S. Sadasivan and K Manikam Hawks Physiological Chemistry : B. L Oser (ed) --------------------------------------------

BC-201 : Human Physiology & Biochemistry of specialized tissues

Biochemistry of blood clotting , clotting factors, intrinsic and extrinsic pathways, mechanism of formation of thrombin, fibrin, fibrin clot, role of vitamin K clotting process, lysis of fibrin clot. Conditions that cause excessive bleeding in humans. Biochemistry of vision, rhodopsin, its desensitization, G protein cascade, visual cycle, colour vision. Biochemistry of muscle contraction, myosin and actin, structure and physiological role, regulation of muscle contraction, role of troponin and tropomyosin, smooth muscle contraction. Principles of gaseous exchange during respiration, transport of oxygen and carbon dioxide in the blood, regulation of respiration. Hormones : types, examples, mechanism of action of steroid and protein hormones, secondary messangers, cAMP, calcium, diacyl glycerol, IP3, regulation gene expression by steroid hormones. Regulation of acid-base balance, types and functions of acid-base buffers, clinical abnormalities associated with acid-base imbalance. Biological membranes and transport, physico-chemical properties of cell membranes, molecular constituents and architecture of membranes, asymmetric organization of membrane lipids and proteins. Ion transport across membranes, types of transport Membrane channels and pumps, acetylcholine receptor channel, ligand and voltage gated channel, generation of action potential during nerve impulse transmission. Active transport, Na/K pump, structure, mechanism of action and significance.

Recommended Books : 1. 2. 3. 4. Molecular Cell Biology : Darnell, Lodish and Baltimore Biochemistry : L. Stryer Lehninger Principles of Biochemistry : D. L. Nelson and M.M. Cox Text Book of Medical Physiology : A.C. Guyton ----------------------------------

BC-202 : Analytical Biochemistry

Principle of pKa, buffer and pH , molarity and normality Basic principle and biological applications of ion exchange, gel filtration, partition, affinity, gas, thin layer, paper and high performance liquid chromatography (HPLC). Principle, instrumentation and application of visible and UV spectroscopy. Dialysis and ultrafiltration. Ultracentrifugation (velocity and buoyant centrifugation, molecular weight determination. density), density gradient

Basic technique of poly acrylamide/starch/ agarose gel electrophoresis, use of SDS and urea, isoelectrofocussing, fulse field gel electrophoresis. Principles and applications of tracer techniques in biology, radioactive isotopes, autoradiography, Cerenkov radiation, liquid scintillation counter. Principles and applications of IR, NMR, Atomic absorption and X-ray diffraction spectroscopy. Principle and applications of microscopy (SEM and TEM). phase contrast, fluorescence and electron

Recommended Books 1. 2. 3. 4. 5. Physical Biochemistry : D. Freifelder Tools in Biochemistry : D. Cooper Analytical Biochemistry : D. Holmes and H. Peck Biophysical Chemistry : Upahye and Upadhye Physical Biochemistry : K.E. VanHolde

------------------------------BC-203 : Immunology
An Introduction: General properties of immune responses innate and adaptive immunity, types of adaptive immune responses, cardinal features of adaptive immune responses, cellular components of adaptive immune system, phases of adaptive immune response (recognition of antigens, activation of lymphocytes, elimination of antigens, decline of immune responses).

Cells and Tissues of the immune system: Lymphocytes, antigen presenting cells , bone marrow , thymus , lymph nodes and lymphatic system , spleen , cutaneous immune system , mucosal immune system. Antibodies and Antigens: Natural production and distribution of antibodies , molecular structure of antibodies , antibody binding of antigens (features of biologic antigens, structural and chemical basis of antigen binding ) , structure-function relationship in antibody molecules (features related to antigen recognition and effecter functions). Major Histocompitability Complex: Discovery of MHC (mouse and human) , properties of MHC genes , structure, properties and cellular distribution of MHC molecules , binding of peptides to MHC ,molecules , genomic organization of MHC , expression of MHC molecules , immune responsiveness . Lymphocyte maturation and activation: General features of lymphocyte maturation early maturation , acquisition of functional competence , maturation of B and T lymphocytes ,general features of T lymphocyte activation , functional responses of T lymphocyte . B cell activation and antibody production General features of humoral immune responses: antigen recognition and antigen induced B cell activation , helper T cell dependent antibody responses to protein antigens , antibody responses to T cell independent antigens , regulation of humoral immune responses by Fc receptors. Innate immunity: Features of innate immune recognition, components of innate immune system , role of innate immunity in local and systemic defence against microbes and in stimulating adaptive immune responses Immunologic tolerance: General features of immunologic tolerance , T and B lymphocyte tolerance , tolerance induced by foreign protein antigens , termination of normal immune response Immune system and health: Immune response to infectious diseases (viral , bacterial and protozoal ), Immuno defficency disorders (SCID ,CVI, AIDS) Cancer and immune system Transplantation immunology Vaccines : types and applications

Recommended Books : 1. 2. 3. 4. 5. 6. 7. Immunology : J. Kuby , R. Goldsby ,T. Kindt , B. Osrorne Fundamental immunology : R. Coleman , M. Lombard , R. Sicard Cellular and Molecular Immunology : A. Abbas , A. Lichtman Immunology for Life Sciemtist : Lesley Jane Eales Immunology : K.R.Joshi, N. O. Osamo Introduction to immunology : J. W. Kimball Immunology : D. M. Weir, J. Steward

-------------------------------

BC-204 : Lab. Course II

1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. Purification of a typical enzyme (beta amylase) Effect of temperature, activators and inhibitors on the enzyme activity Determination of Km and Vmax of amylase by double reciprocal plot Determination of specific activity and turn over number of salivary amylase Immobilization of typical enzyme Concept of pH, buffers, molarity and normality Colorimetric estimation of protein Colorimetric estimation of inorganic phosphate Separation of amino acids by paper chromatography Separation of plant secondary metabolites by TLC Purification of proteins by ion-exchange chromatography Paper electrophoresis Determination of molecular weight of protein by gel permeation chromatography (Demonstration) 14. Demonstration of HPLC / HPTLC / PAGE 15. Demonstration of GC / AAS / Lyophilizer Recommended Books : 1. 2. 3. 4. 5. 6. An introduction to practical biochemistry : Plummer, D A laboratory manual in biochemistry : Jayaraman, J. Practical Biochemistry : K Wilson and Walker Biochemical Methods : S. Sadasivan and K Manikam Hawks Physiological Chemistry : B. L Oser (ed) Experimental Biochemistry; A student companion : B. S. Rao and V. Deshpande

---------------------------------BC-205 : Lab. Course - III

1. Estimation of total cholesterol, LDL and HDL. 2. Estimation of SGOT and its pathological significance. 3. Estimation of SGPT and its pathological significance. 4. Determination of serum alkaline phosphatase 5. Urine examination: normal & abnormal constituents 6. Estimation of glucose in blood by enzymatic method 7. Estimation of serum bilirubin 8. Determination of clotting and prothrombin time 9. Determination of blood groups 10. Widal Test 11. Quantitative estimation of antigen sandwitch ELISA method (antibody capture method) 12. ELISA (qualitative method) 13. Radial Immuno diffusion 14. Western blotting (D)

10

15. Estimation of Thyroid hormones by RIA (D) Recommended Books 1. A Handbook of Practical and Clinical Immunology : G.P. Talwar and S. K. Gupta (Eds) 1. Hawks Physiological Chemistry : B.L. Oser (ed) 2. Practical clinical biochemistry : Varlley 4. Experimental Biochemistry; A student companion: B. S. Rao and V. Deshpande

------------------------------------

11