M. Sc Biotech 2010 JNTUH

CENTRE FOR BIOTECHNOLOGY

INSTITUTE OF SCIENCE AND TECHNOLOGY

JAWAHARLAL NEHRU TECHNOLOGICAL UNIVERSITY HYDERABAD

Kukatpally, Hyderabad-500 072, Andhra Pradesh, INDIA

M. Sc.

In

BIOTECHNOLOGY

(FTPG)

COURSE STRUCTURE

&

DETAILED SYLLABUS

EACH SEMESTER IS APPROXIMATELY 20-21 WEEKS DURATION.

INCLUDING EXAMINATIONS.

EACH PERIOD IS ABOUT 50 MINITUES DURATION.

THERE WILL BE NORMALLY FOUR SESSIONS PER DAY EACH OF

ABOUT 100 MINITUES DURATION

TWO SESSIONS OF LABORATORY IS EQUIVALENT TO ONE SESSION

OF THEORY

1

M.Sc. (BIOTECHNOLOGY)

COURSE STRUCTRURE

I YEAR

I SEMESTER

SUBJECT

CODE

SUBJECT TITLE PERIOD

/WEEK

CREDITS

MBT-101 Cell Biology 4 3

MBT-102 Biomolecules 4 3

MBT-103 Computer Applications and Biostatistics 4 3

MBT-104 Microbial Physiology and Genetics 4 3

MBT-105 Molecular Biology 4 3

MBT-106 Basic Engineering Mathematics 4 2

MBTL-11 Cell Biology Lab 3 1

MBTL-12 Biomolecules Lab 3 1

MBTL-13 Computer Applications and Biostatistics Lab 3 1

MBTL-14 Microbial Physiology and Genetics Lab 3 1

MBTL-15 Molecular Biology Lab 4 1

TOTAL: 40 22

II SEMESTER

SUBJECT

CODE


/WEEK

CREDITS

MBT-201 Macromolecules and Analytical Techniques 4 3

MBT-202 Biology of the Immune System 4 3

MBT-203 Process Engineering Principles 4 3

MBT-204 Enzyme Technology 4 3

MBT-205 Environmental Biotechnology 4 3

MBTL-21 Macromolecules and Analytical Techniques Lab

4 2

MBTL-22 Biology of the Immune System Lab 4 2

MBTL-23 Process Engineering Principles Lab 4 1

2

MBTL-24 Enzyme Technology Lab 4 1

MBTL-25 Environmental Biotechnology Lab 4 1

TOTAL: 40 22

II YEAR

III SEMESTER

SUBJECT

CODE


/WEEK

CREDITS

MBT-301 Genetic Engineering 4 3

MBT-302 Plant Biotechnology 4 3

MBT-303 Animal Cell Science and Technology 4 3

MBT-304 Bioprocess Engineering and Technology 4 3

MBT-305 Bioinformatics 4 3

MBTL-31 Genetic Engineering Lab 4 2

MBTL-32 Plant Biotechnology Lab 4 2

MBTL-33 Animal Cell Science and Technology Lab 4 1

MBTL-34 Bioprocess Engineering and Technology Lab 4 1

MBTL-35 Bioinformatics Lab 4 1

TOTAL: 40 22

IV SEMESTER

SUBJECT

CODE


/WEEK

CREDITS

MBTP-301 Research Project 22 22

TOTAL: 22 22

3

M.Sc. Biotechnology, I semester

MBT-101 CELL BIOLOGY

UNIT I: CELL STRUCTURE AND FUNCTION

Diversity of cell size and shape; Cell theory; Structure of Prokaryotic and Eukaryotic

cells; Cellular organelles and their organization; Cell walls-of eubacteria (peptidoglycan)

and related molecules; Outer-membrane of Gram- negative bacteria; Cell wall and cell

membrane synthesis; Cell inclusions like endospores, gas vesicles; Cell motility- cilia,

flagella of eukaryotes and prokaryotes, Microscopic techniques for study of cells, Sub-

cellular fractionation and criteria of functional integrity.

UNIT: II CELL MEMBRANE TRANSPORT AND ENERGY TRANSDUCTIONS

Transport of nutrients, ions and macromolecules across cell membranes.

Cellular energy transactions- role of mitochondria and chloroplast

UNIT:III CELL CYCLE AND SIGNALING

Cell cycle- Various phases and checkpoints of cell cycle and role in tumor formation.

Mitosis and Meiosis.

Cell signaling- overview and different types of cell signaling, Major receptor of cell

signaling, Mechanisms of signal transduction.

Biology of cancer- Types and causes, classification of tumor, Development of cancer.

UNIT:IV PROTEIN SYNTHESIS AND METABOLIC PATHWAYS

Biosynthesis of proteins in Eukaryotic cell, Co and post-translational modification

intracellular protein traffic.

Metabolite pathways and their regulation- Genetic regulation, Control of enzyme activity,

regulation of biosynthetic pathways, regulation of degradative pathways, Regulation by

feed back inhibition.

4

UNIT:V CELL DIFFERENTIATION AND MOLECULAR BIOLOGY OF

PATHOGENS

Cellular basis of differentiation and development - Gametogenesis and fertilization,

Development in Drosophila and Arabidopsis; Spatial and temporal regulation of Gene

Expression.

Brief introduction to the Life Cycle and Molecular Biology of some important pathogen

of AIDS, Malaria, Hepatitis, Tuberculosis, Filaria.

PRACTICALS:

1. Microscopy: Compound Microscope

2. Motility of bacteria

3. Protoplast isolation

4. Mitosis of onion root tip.

5. Isolation and estimation of Chloroplast

6. Mitochondria isolation and study of biochemical activity.

7. Cell signaling , Banding , Karyotyping

BOOKS:

1. Molecular Biology of cell, Alberts. B et al.

2. Molecular Cell Biology, Lodish et al.

3. Reproduction in Eukaryotic cells, DM Prescott, Academic press.

4. Developmental Biology, SF Gilbert, Sinauer Associates Inc.

5. Cell in Development and inheritance, EB Wilson, MacMilan, New York.

6. The Coiled Spring, Ethan Bier, Cold Spring Harbor Press.

7. Fertilization, FT Longo, Chapman and Hall

5


MBT-102 Biomolecules

UNIT I: Fundamentals of biochemistry

Water, pH, pK, buffers, covalent bond, non-covalent interactions.

Bioenergetics: free energy, enthalpy, entropy, laws of thermodynamics, high energy

compounds.

UNIT II: Biomolecules

Classification, physical and chemical properties of carbohydrates, lipids, aminoacids and

proteins; protein structral hierarchy, ramachandran plot; nucleotides and nucleic acids.

Pigments and storage materials.

UNIT III: Enzymes

Classification, mechanisms of action of enzymes, enzyme assay methods, michaelies-

menten kinetics, inhibition of enzyme catalysis, regulatory enzymes. Factors effecting

enzyme activity.

UNIT IV: Photosynthesis

Photosynthesis- oxygenic and anoxygenic, central photochemical events.

UNIT V: Metabolism

Catabolism of biomolecules – Carbohydrates- aerobic and anaerobic fate of

carbohydrates, E.T.C,

Lipids – oxidation of fatty acids – saturated and unsaturated, fatty acids with odd no of

carbon atoms.

Proteins – deamination, transamination aminoacids – eg; L-Aspargine, L-valine, L-

phenyl alanine.

Anabolism of biomolecules – carbohydrates – synthesis of carbohydrates, HMP shunt,

Biosynthesis of lipids, Biosynthesis of aminoacids – Glutamate, Tyrosine, Proline.

6

PRACTICALS:

1. Titration of amino acids

2. Determination of pK

3. Reactions of amino acids, sugars and lipids- quantitative reactions

4. Analysis of oils-iodine number, saponification value, acid number.

5. UV, Visible, Absorption spectra.

6. Centrifugation, SDS-PAGE & TLC –Silver staining

7. Estimation of inorganic phosphorus by Fiske Subbarow Method

8. Isolation and estimation of protein using various colorimetric and spectrophotometric

methods.

BOOKS

1. Biochemical Calculations, lrwin H. Segel, John Wiley and Sons Inc.

2. General Chemistry. Linus Pauling, W.H. Freeman & Company.

3. Organic Chemistry, DJ Cram and GS Hammond, McGraw Hill.

4. Biochemistry. D Voet and JG Voet, J Wiley and Sons.

5. Physical Biochemistry, D Freifilder, W.H. Freeman & Company.

6. Laboratory Techniques in Biochemistry and Molecular Biology, Work and Work.

7. Understanding Chemistry, CNR Rao, Universities Press. Hyderabad 1999.

8. A Biologist’s Guide to Principles and Techniques of Practical Biochemistry, K

Wilson & KH Goulding, ELBS Edition, 1986.

9. Tools of Biochemistry by T.G. Cooper.

7


MBT-103 COMPUTER APPLICATIONS & BIOSTATISTICS

UNIT-I : Introduction of digital computers: Organization, low and high level languages-

Computer Basics, Computer Capabilities, Types of computers, Central Processing Unit,

functions of ALU and control unit (CU), Operating system-Microsoft – Office

packages(MS-Word, MS-Excel, MS-Power Point),Algorithm, Flowchart, Program

development steps

UNIT-II :, Basic Structures of C language, C tokens, Data types, declaration of

variables, assigning values, arithmetic, relational and logical operator, increment and

decrement operators, control operator, bit-wise operator, expressions, evaluation, input-

output operators. IF, SWITCH statement, WHILE, DO-WHILE and FOR statements,

One ,Two dimensional arrays, initialization, functions, string variables, declaration,

reading, writing, string handle function, user-defined functions, variables & storage

lasses,. Structures, unions, pointers.

Unit-III : Introduction to statistics :population-sample –primary data and secondary data

- graphical and diagrammatic representation of data- Measure of central tendency-Mean,

median and mode-measure of dispersion-range-standard deviation -raw and central

moments-skewness and kurtosis(definition only)-Concept of probability –classical and

relative frequency definition of probability-addition and multiplication laws of

probability( without proofs) and examples.

Unit-IV : Concept of Random variables: Probability mass function-probability density

function-probability distribution function( definitions only)- Binomial ,Poisson and

Normal distribution( definitions and statements of properties and examples).Principles of

least square-fitting of straight line-parsons coefficient of correlation and concept of linear

regression.

8

Unit-V: Concept of testing of hypothesis critical region-two type errors-level of

significance of large sample tests for single mean and difference of means. single

proposition and difference proportion- chi square test for goodness of fit and chi square

test of means and f-test for equality of variances-arithmetic, Geometric and harmonic

mean.

BOOKS

1) Statistical methods S.P.Gupta. S Chand Pubplications

2) Business Statistics by S.P Gupta & M.P.Gupta

3) Programming in ANSI C by E.Balaguruswami

9


MBT-104 MICROBIAL PHYSIOLOGY AND GENETICS

UNIT I: Methods in Microbiology: Pure culture techniques; Theory and practice of

sterilization; Preservation and maintenance of cultures and Culture collection centres;

Principles of microbial nutrition Construction of culture media; Enrichment culture

techniques for isolation of chemoautotrophs, chemoheterotrophs and photosynthetic

microorganisms, Enrichment culture methods,

UNIT II: Overview of Microbial Metabolism

UNIT III: Microbial Genetics : Methods for Exchange of genetic material -

Transformation, Transduction, Conjugation; Plasmids, episomes, Bacterial DNA etc.

UNIT IV: Archaea: archaea; Thermoplasma; Eukarya: Algae, Fungi, Slime molds and

Protozoa; Viruses: structure of viruses; Lysogeny and lytic cycles of phage , DNA

viruses; Positive strand, Negative strand and double stranded RNA viruses; Replication;

Examples of Herpes, Pox, Adenoviruses, Retroviruses, Viroids and Prions,.

UNIT IV: Microbial Diseases & Chemotherapy/Antibiotics: Disease reservoirs;

Epidemiological terminologies; Infectious disease transmission; Respiratory infections

caused by bacteria and viruses; Tuberculosis; Sexually transmitted diseases including

AIDS; Diseases transmitted by animals (rabies, plague), insects and ticks (rickettsias,

lyme disease, malaria), Food and water borne diseases; Public health and water quality;

Pathogenic fungi; Emerging and resurgent infectious diseases.

10

PRACTICALS:

1) Growth Curve of bacteria

2) Factors effecting the growth of microorganisms

3) Determination of thermal death point

4) Determination of antimicrobial activity (Disc diffusion, cup plate method)

5) Determination of minimum inhibitory concentrations

6) Bacterial Transduction Generalised/Specialised

7) Phage lysogeny and lytic cycle using E.coli strains

8) Transfection -GFPs

BOOKS:

1. Microbiology, Michael J Pelczar Jr, E.C.S.Chan and Noel R. Krieg, Tata McGraw-

Hill Edition (5th Edition)

2. General Microbiology 5th Edition Stanier et al.

3. Text of Microbiology, D.K.Maheshwari and R.C.Dubey, S. Chand Publication

Reprint 2009

4. Ananthanarayan and Paniker’s Text of Microbiology, Edited by C K J Paniker

University Press (3rd Edition) 2008

11


MBT-105 MOLECULAR BIOLOGY

THEORY:

UNIT-I: Information carrier molecules- DNA & RNA: Experimental evidences to

prove DNA as hereditary genetic material, DNA- structures (A,B,Z types), DNA

denaturation & renaturation kinetics, Super coiled DNA features, Experimental

evidences to prove RNA as hereditary genetic material, Types of RNA- mRNA, tRNA,

rRNA. Organelles DNA (Chloroplast/ Mitochondria).

UNIT – II: DNA Replication: Modes of replication, DNA polymerases (Prokaryotic

& Eukaryotic), Unidirectional, bidirectional & rolling circle replication, Leading and

lagging strand synthesis, Topoisomerases, Telomeric replication, Inhibitors of DNA

replication

DNA damage and repair- Types of DNA damage repair mechanisms-

Photoreactivation, Excision repair, Recombination repair, Error prone (SOS) repair,

Relationship between DNA damage & cell cycle regulation

UNIT-III: Gene expression-Transcription Transcription (Prokaryotic & Eukaryotic),

Promoters, Eukaryotic Enhancers, RNA polymerases, Sigma factor in prokaryotes,

Transcription steps – initiation, elongation, termination. Transcription inhibitors,

Processing of tRNA and Rrna, Post translational modifications-splicing mechanisms.

RNA editing.

UNIT- IV: Gene Expression – Translation Prokaryotic & Eukaryotic Ribosome,

Genetic code, Wobble Hypothesis, Translation steps - initiation, elongation, termination.

Translation inhibitors.

UNIT V:

Transcriptional level gene regulation in prokaryotes & Eukaryotes, Operons.

12

PRACTICAL:

1. Chromosomal DNA Isolation and quantification

2. Plasmid DNA Isolation and quantification

3. Electrophoresis of nucleic acid DNA & RNA

4. Bacterial Conjugation

5. Bacterial Transformation

6. RNA isolation.

BOOKS

1. Molecular Cloning:A Laboratory Manual, J. Sambrook, E.F. Fritsch and T Maniatis,

ColdSpring Harbor laboratory Press, New York, 2000

2. Introduction to Practical Molecular Biology, P.D. Dabre, John Wiley & Sons Ltd,,

New York, 1988

3. Molecular Biology LabFax, T.A. Brown (Ed.), Bios Scientific Publishers Ltd.,

Oxford, 1991

4. Molecular Biology of the Gene (4th Edition), J.D. Watson, N.H. Hopkins, J.W.

Roberts, J.A.Steitz and A.M. Weiner, The Benjamin/Cummings Publ. Co., Inc.,

California, 1987.

5. Molecular Cell Biology (2nd Edition) J. Darnell, H. Lodish and D. Baltimore,

Scientific American Books, Inc., USA, 1994

6. Molecular Biology of the Cell (2nd Edition) B. Alberts, D. Bray, J. Lewis,M. Raff, K.

7. Roberts, and J. D. Watson. Garland publishing, Inc., New York, 1994

8. Gene VI (6th Edition) Benjamin Lewin, Oxford University Press, U.K., 1998, VCH

Publishers, Inc., New York, 1995

9. Genomes, T.S. Brown

13


MBT- 106 BASIC ENGINEERING MATHEMATICS

UNIT I:

Quadratic expression – Quadratic equation, roots of quadratic equation of the form

ax2+bx+c=0 and simple properties of quadratic roots. Numerical problem solving on

Quadratic equation

Theory of equations- polynomial function, polynomial equation, reminder theorem,

synthetic division, to find the coefficient and reminder of polynomial function, relation

between the roots and coefficients of f(x) = 0

UNIT II:

Trigonometry- relations related to compound angles, multiple and sub-multiples,

transformations, hyperbolic functions

UNIT III

Concepts of limit, continuity, differentiation, product rule, quotient rule. Differentiation

of trigonometric, logarithmic, exponential functions.

Applications of differentiation – problems on tangent, sub tangent normal, sub normal.

Introduction to partial differentiation, Euler’s theorem.

UNIT IV:

Introduction, Integration of different functions, methods of Integration, Integration by

parts. Concept of definite integrals.

Applications of definite integrals – problems on areas.

UNIT V:

Forming of differential equation by eliminating arbitary constants, first order and first

degree – variables and seperables, exact, homogeneous and linear.

Textbooks:

1. Engineering Mathematics - N.P. Bali and others.

2. Engineering mathematics - B.V. Ramana

References:

1. Differential Calculus - Shanthi Narayan

2. Integral Calculus - Shanthi Narayan

14

M.Sc. Biotechnology, II semester

MBT-201 MACROMOLECULES AND ANALYTICAL TECHNIQUES

UNIT I: Macromolecules and supra molecular assemblies- Types of macromolecules in

biological systems, molecular assemblies like membranes. ribosomes, extracellular

matrix. Examples of Supramolecular assemblies viral caspids.

UNIT II: Microscopy: Electron Cryomicroscopy; Scanning Tunneling microcopy, X-ray

Crystallography, Ultra centrifugation, Spectroscopy techniques: UV, IR, NMR, LASER

Raman Spectroscopy, Florescence Spectroscopy, Differential colorirnetry

UNIT III: Radiochemical methods: radioactivity, measurement of radioactivity,

photographic emulsion, ionisation chamber, Gieger Muller counter, autoradiography.

UNIT IV: Sequencing of proteins and nucleic acids. Protein-protein and protein-ligand

interactions, physical and chemical methods for study, 2 D- Isoelectric focusing.

Conformational properties of polynuleotides and polysaccharides - secondary and tertiary

structural features and their analysis- theoretical and experimental; protein folding-

biophysical and cellular aspects.

UNIT V : Chromatography – adsorption, affinity, partition, Ion exchange, gel

permeation, GLC, TLC, RPC, HPLC etc. Electrophoresis: Different methods of

electrophoresis for protein, nucleic acids, small molecular weight compounds and

immuno precipitates (Immuno electrophoresis). Peptide mapping and combination of

electrofocussing and SDS-PAGE.

PRACTICALS

1. Electrophoresis of Proteins - native and under denaturing conditions –silver staining, comaise staining.

2. Separation techniques (HPLC, GPC, FPLC, Ion-Exchange).

3. Determination of Tm of nucleic acid.

15

4. Isoelectric focusing (Futute)

BOOKS

1. Essentials of Molecular Biology, David Friefilder, Jones and Barllett Publications.

2. Proteins-Structure and Molecular Properties. TE Creighton, WH Freeman and

company.

3. Genes VII, B. Lewin, Oxford University Press.

4. Introduction to Protein Structure, C. Branden and J. Tooze, Garland Publishing, New

York.

5. Encyclopaedia of Molecular Biology, J. Kendrew, Blackwell Scientific Publications,

Oxford.

6. Physical Chemistry of Macromolecules, Tanford, C., John Wiley and Sons.

7. Introduction to Biophysical Chemistry, RB Martin, MeGraw Hill, New York.

8. Biophysical Chemistry, Cantoz, WH Freeman.

9. Protein Structure, by Max Perutz.

16


MBT-202 BIOLOGY OF THE IMMUNE SYSTEM

UNIT I: Introduction -Phylogeny of Immune System - Innate and acquired immunity -

Clonal nature of immune response, Humoral and Cell mediated immunity, Nature and

Biology of antigens and super antigens.

UNIT II: Organization and structure of lymphoid organs: Lymphoid follicle, Thymus

Lymph node, Spleen, MALT, CALT, SALT. Skin associatetd Lymphoid Tissue.

UNIT III: Cells of the immune system: Hematopoiesis and differentiation, Inflammation

(IL6), Macrophages, Dendritic cells, Natural killer and Lymphokine activated killer cells,

Eosinophils, Neutrophils and Mast-Cells (IL2)

UNIT IV: Immunochemistry: Antibody structure and function, Hybridoma Technology

and Monoclonal antibodies, Antigen- antibody interactions

Humoral Immunity: BCR and generation of Immunoglobulin diversity, Activation of B-

Lymphocytes, Generation of humoral immune responses

Cell mediated Immunity: Major histocompatibility complex, MHC restriction, Antigen

processing and presentation, TCR, generation of diversity, generation of cell mediated

immune responses, Mechanism of T cell and NK cell mediated lysis, Antibody dependent

cell mediated cytotoxicity , IL2.

UNIT V: Immunopathogenesis of Autoimmune disorders, Transplantation, Vaccination

for disease control. Common disease and control through Vaccination.

PRACTICALS

1. Radial immuno diffusion

2. Ouchterolony double immuno diffusion

3. Dot blot

17

4. Immuno electrophoresis

5. Quantitative precipitin assay

6. Purification of antibodies

7. Lateral flow diagnosties – HCG, malaria, Tuberculosis.

8. Blood grouping – Agglutination.

9. Cell identification techniques based on staining

Text Books:

1. Kuby Immunology (Kindt, Kuby Immunology) - Thomas J. Kindt, Barbara A.

Osborne, Richard A. Goldsby, publisher: W. H. Freeman, 2006

2. Immunology- David Male, Jonathan Brostoff, David Roth, Ivan Roitt, publisher:

Mosby, 2006

Reference Books:

1. Fundamental Immunology- William E Paul, publisher: Lippincott Williams &

Wilkins, 2008

2. Immunology, Infection, and Immunity - Gerald B. Pier, Jeffrey B. Lyczak, Lee

M. Wetzler, publisher: ASM Press, 2004

3. Lecture Notes: Immunology, 5th Edition- Ian Todd, Gavin Spickett, publisher:

Wiley-Blackwell, 2005

4. Immunology: A Short Course- Richard Coico, Geoffrey Sunshine, publisher:

Wiley-Blackwell, 2009

5. Cellular and Molecular Immunology- Abul K. Abbas MBBS, Andrew H.

Lichtman MD PhD, Shiv Pillai MD, publisher: Saunders, 2007

18

6. Roitt's Essential Immunology (Essentials) - Peter Delves, Seamus Martin,

Dennis Burton, Ivan Roitt, publisher: Wiley-Blackwell, 2006

7. Schaum's Outline of Immunology- George Pinchuk, publisher: McGraw-Hill,

2001


MBT-203 PROCESS ENGINEERING PRINCIPLES

UNIT I: Fundamentals o f chemical engineering; Brief overview , concepts of unit

operation & unit process with examples, units and dimensions, dimensional analysis,

Presentation and analysis of data,fluids vs solids, fluid static’s- Pressure measurement –

different types of manometers with simple numeric problems, steady flow –equation of

continuity , Derivation of Bernoullis equation for one dimensional flow, its limitation and

corrections and applications in flow meters.

UNIT II: Fluids mechamics; Introduction , Newtons law of viscosity, flow curves for

non-Newtonian fluids with examples from biotechnology, flow patterns –laminar and

turbulent, Reynolds experiment – Reynolds number, pressure drop due to friction,

relating Reynolds number with friction factor, packed beds and pressure drop through

packed beds ,Fludization – concept , significance in chemical industry, and pressure drop

across fluidized beds.

UNIT III: Heat transfer; Modes of heat transfer with examples, conduction, convection

and radiation, definitions and governing laws, Thermal boundary layer and Prandtal

number, conductivity and resistivity, compound resistances in series, heat transfer

through cylindrical pipe, analogy between momemtum and heat transfer, simple

numerical problems on conduction.

Convection-Natural and forced convection, dimensionless numbers in heat transfer,

concept of heat transfer coefficient and overall heat transfer coefficient, radiation,

combined heat transfer by conduction, convection and radiation.

19

UNIT IV : Heat exchanger equipment: Heat exchangers Over view of heat

exchangers-types and temperature area graphs, concept of LMTD, overview of other heat

transfer operations.

Boiling and evaporation; Mechanisms of boiling, Film and nucleate boiling, single and

multiple effect evaporators, economy and evaporators performance with various feeding

patterns viz, forward , backward and parallel.

UNIT V: Mass transfer Definition ,Ficks law of diffusion , Analogy with momemtum

and heat transfer, diffusivities of gases and liquids, fundamentals of mass transfer ,

theories of mass transfer, concept of mass transfer coefficients, dimesionless numbers,

overview of separation operations with examples; ideal stage concept, mass transfer

equipment-distillation, liquid extraction, gas absorption, drying.

TEXT BOOKS

1) Unit operations of Chemical Engineering, (1993), by McCabe, Simth and Harriott,

McGraw Hill publishers.

2) Introduction to Biochemical Engineering, Second edition, By D G Rao, Tata McGraw

Hill Publications.

3) Bioprocess Engineering principles By Pauline M Doran, Academic Press.

20


MBT-204 ENZYME TECHNOLOGY

THEORY

UNIT I: Overview of intermediary metabolism Fundamentals of enzymes: Chemical

Nature of enzymes, enzyme nomenclature, sources of enzymes, Isolation of enzymes

from different sources, Screening for novel enzymes, media for enzyme production,

Enzyme assay methods, purification of enzymes, preparation of enzymes, industrially

important enzymes, safety and regulatory aspects of enzyme use.

UNIT II: Enzymology Enzyme catalysis- active site, substrate, transition state,

activation energy, binding energy, enzyme specificity, Thermodynamics of enzyme

catalysis; mechanism of enzyme action- covalent catalysis, metal ion catalysis, general

acid-base catalysis, Induced-fit, proximity and orientation, factors effecting enzyme

activity.

UNIT III: Metabolism Enzyme kinetics- First, second, zero and pseudo-order kinetics;

Prs-steady and steady-state kinetics; Derivation of Michaelis-menten equation, Km, V max,

turnover number, catalyte efficiency, specificity constant, linear transformation to M.M

equation, kinetics of bi-substrate reactions, Random and sequential order, Ping-pong

mech, Differentiation of different mechanism of bi substrate reactions.

UNIT IV: Regulatory enzymes- Allostiric enzymes- MWC model and KNF model,

kinetic properties of allosteric enzymes; nemonical enzymes and enzyme memory.

Enzyme Inhibition; Reversible and irreversible inhibition and kinetic properties.

UNIT V: Isoenzymes, Multienzyme complexes and multifunctional enzymes.

Structure and activity of the enzymes Mechanism of action of chymotrypsin,

glyceraldehyde 3 Phosphate dehydrogenase, lysoenzyme, carboxy peptidase,

ribonuclease, aldolase etc. Various techniques used for the immobilization of

21

enzymes Applications of immobilized enzyme in Biotechnology. Enzyme activity in non-

aqueous media and then industrial applications.

Riboenzyme and catalytic antibodies- Functional proteins- structure and drug targets

(enzymes and receptors)

TEXT BOOKS :

1. "Biochemical Engineering fundamentals" by J E Bailey and D F Ollis, 2nd ed,

McGraw-Hill (1986).

2. Enzymes: Biochemistry, Biotechnology, Clinical Chemistry by T. Palmer Horwood

Publications 2001.

PRACTICAL

1. Estimation of acid phosphatase from plant/animal sources.

2. Enzyme concentration by salt fractionation.

3. Determination of optimum pH, temperature, Km etc of the enzyme.

4. Extraction of enzyme

5. Dialysis

6. Column Chromatography

7. Activity staining for enzyme

8. Enzyme immobilization by entrapment

22


MBT-205 ENVIRONMENTAL BIOTECHNOLOGY

UNIT I: Ecosystem And Biodiversity Ecosystem structure and function. Types of

ecosystem; Aquatic and terrestrial Values of biodiversity, Threats to biodiversity, various

methods of Conservation of biodiversity, Red Data Book

UNIT II:Environmental Pollution And Global Issues Environmental Pollution:

Types of pollution, Air Pollution, Water Pollution, their sources, effects and control

methods. Global environmental issues: Green House Effect, Global Warming, Acid

Rain, Ozone Depletion and their impact on environment.

UNIT III: Waste Water Treatment Technologies Waste water types, Major

contaminants in waste water. Physical, chemical and biological methods of waste water

treatment.Aerobic; Activated Sludge Process, Trickling Filters, Biological Filters,

Rotating Biological Contractors, Fluidized Bed Reactor.

Anaerobic; Contact Digesters, Packed Column Reactors, UASB biological treatment

process.

UNIT IV: Bioremediation And Phytoremediation Definition constraints and priorities

of Bioremediation, Types of bioremediation, In-situ and Ex-situ bioremediation

techniques, Factors affecting the use of bioremediation. Phytoremediation, advantages

and limitations, various phytoremediation processes; Phytoextraction, phytostabilization,

phytotrnsfermation.

UNIT V Environmental Management

Solid waste management.Development of biodegradable and ecofriendly products:

Biopolymers, Biodesel, Biofertilizers and Biopesticides.

Integrated Pest Management.

Concepts of Environmental Impact Assesment and Environmental Audit

23

PRACTICALS:

1. Biological Oxygen Demand

2. Chemical Oxygen Demand

3. MPN

4. Coliform bacteria

5. Reduction of BOD & COD after incubation with microbes

REFERENCE TEXT BOOKS:

1. Wastewater Engineering-Treatment, Disposal, and Resuse, Metcalf and Eddy,

Inc., Tata McGraw Hill, New Delhi.

2. Industrial Pollution control Engineering- AVN Swamy., Galgotia Publication,

(2006).

3. Environmental Biotechnology- Allan Stagg.

24

M.Sc. Biotechnology, III semester

MBT-301 GENETIC ENGINEERING

THEORY

UNIT I: Scope of Genetic Engineering Milestones In Genetic Engineering - Isolation of

enzymes, DNA sequencing, synthesis and mutation, detection and separation, cloning,

gene expression. Cloning and patenting of life forms. Genetic engineering guidelines.

UNIT II:Molecular Tools in genetic engineering: Restriction enzymes, ligases, s1

nuclease, terminal deoxynucleotides, transferases, Poly A polymerases, Reverse

Transcriptase, Alkaline phosphatase etc., modification enzymes, DNA, and RNA

markers.

UNIT III: Nucleic Acid Purification, Yield Analysis. Nucleic Acid Amplification (PCR)

and Its Applications, Restriction Mapping of DNA Fragments and Map Construction,

Nucleic Acid Sequencing, Gene Cloning Vectors; Plasmids, bacteriophages, phagemids,

cosmids, Artificial chromosomes.

cDNA Synthesis and cDNA library preparations, Cloning mRNA enrichment, reverse

transcription, DNA primers, Linkers, adaptors and their chemical synthesis, Library

construction and screening, Genomic libraries (complete sequencing projects)

Alternative Strategies of Gene Cloning Cloning interacting genes- Two-and three hybrid

systems, cloning differentially expressed genes. Site-directed Mutagenesis and Protein

Engineering

UNIT IV: How to Study Gene Regulation? DNA transfection, Northern blot, Primer

extension, S1 mapping, RNase protection assay, Reporter assays. Nucleic acid

microarrays (Protein & NA). Expression Strategies for Heterologous Genes Vector

engineering and codon optimization, host engineering, In vitro transcription and

translation, expression in bacteria, expression in Yeast, expression in insects and insect

cells, expression in mammalian cells, expression in plants. Processing of Recombinant

25

Proteins Purification and refolding, characterization of recombinant proteins, stabilization

of proteins

UNIT V: Phage Display, T-DNA and Transposon Tagging, Role of gene tagging in gene

analysis, T-DNA and transposon tagging, Identification and isolation of genes through T-

DNA or transposon, Transgenic and Gene Knockout Technologies Targeted gene

replacement, Chromosome engineering, Gene Therapy Vector engineering, Strategies of

gene delivery, gene replacement/augmentation, gene correction, gene editing, gene

regulation and silencing.

PRATICALS

1. Isolation of DNA, Restriction digestion & Ligation

2. Screening for recombinants

3. PCR-Amplification of DNA.

BOOKS

1. Molecular Cloning: a Laboratory Manual, J. Sambrook, E.F. Fritsch and T. Maniatis,

Cold Spring Harbor Laboratory Press, New York, 2000.

2. DNA Cloning: a Practical Approach, .M. Glover and B.D. Hames, IRL Press, Oxford,

1995.

REFERENCES:

1. Molecular and Cellular Methods in Biology and Medicine, P.B. Kaufman, W. Wu. D.

Kim and L.J; Cseke, CRC Press, Florida, 1995.

2. Methods in Enzymology vol. 152, Guide to Molecular Cloning Techniques, S.L.

Berger and A.R. Kimmel, Academic Press, Inc. San Diego, 1998

3. Methods in Enzymology Vol 185, Gene Expression Technology, D.V. Goeddel,

Academic Press, Inc., San Diego, 1990

4. DNA Science. A First Course in Recombinant Technology, D,A. Mickloss and G.A.

Froyer. Cold Spring Harbor Laboratory Press, New YorK, 1990.

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5. Molecular Biotechnology (2nd Edn.), S.B. Primrose. Blackwell Scientific Publishers,

Oxford, 1994

6. Milestones in Biotechnology. Classic papers on Genetic Engineering, J.A. Davies and

W.S. Reznikoff, Butterworth-Heinemann, Boston, 1992.

7. Route Maps in Gene Technology, M.R. Walker and R. Rapley, Blackwell Science

Ltd., Oxford, 1997.

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MBT-302 PLANT BIOTECHNOLOGY

THEORY

UNIT I: Introduction to cell and tissue Culture. Tissue culture media. Totipotency,

initiation and maintenance of aseptic cultures. Micropropagation, Organogenesis, somatic

embryogenesis. Shoot-tip culture: Rapid clonal propagation. Callus and Cell Suspension

cultures.

Protoplast isolation, culture and fusion; selection of hybrid cells and regeneration of

hybrid plants.

UNIT II: Anther, pollen and ovary culture for production of haploid plants and

homozygous lines. Cryopreservation, slow growth and DNA banking for germ plasm

conservation, Somaclonal Variation. Production of plant secondary metabolites in vitro,

commercial production of secondary metabolites. Technology for yield enhancement, cell

immobilization, , Transformed organ cultures (hairy roots, shooty teratomas).

UNIT III: Plant Transformation technology (Direct, Indirect), Agrobacterium based

vectors, Viral vectors, Different types of promoters, Selectable markers (Positive and

Negative), Reporter genes. Chloroplast Transformation.: advantages. Application of

Plant Transformation for productivity and performance (Biotic Stress: Herbicide

resistance, Insect resistance, Virus Resistance, Disease resistance, Nematode resistance)

Application of Plant Transformation for productivity and performance (Abiotic Stress:

drought, salinity, temperature, oxidative), Quality traits, Post harvest losses, Long shelf

life, Male sterility.

UNIT IV: Secondary metabolite pathway engineering and Molecular Farming for

Pharmaceuticals and Industrial products (plantibodies, edible vaccines, industrial

enzymes, therapeutic proteins, lysosomal enzymes, purification strategies, oleosin

partitioning technology).

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UNIT V: Conventional Plant Breeding, limitations, Molecular Marker-aided Breeding:

RFLP maps, RAPD markers, STS, microsatellites, SCAR (sequence characterized

amplified regions), SSCP (single strand conformational polymorphism), AFLP, Map

based cloning, molecular marker assisted selection.

PRACTICAL

1. Preparation of media.

2. Surface sterilization.

3. Organ Culture.

4. Callus propagation, organogenesis, transfer of plants to Soil.

5. Cytological examination of regenerated plants.

6. Agrobacterium culture, selection of transformants, reporter gene (GUS) assays.

7. RAPD Analysis

8. Extraction & Quantification of Secondary Metabolitess.

BOOKS:

1. Bhojwani SS and Rajdan MK . Plant Tissue Culture : Theory and Practice, A revised

Edition. 2002

2. Hammond, R McGarvey and V. Yusibov (Eds.): Plant Biotechnology. Springer

Verlag, 2000

3. Glick BR and Pasternak JJ . Molecular Biotechnology principles and applicationsof

Recombinant DNA. 2006

REFERENCES:

1. H.S. Chawla: Biotechnology in Crop Improvement. International Book Distributing

Company, 1998.

2. Giri CC and Archana Giri : Plant Biotechnology Practical Manual . I K International

2007

3. R.J. Henry: Practical Application of Plant Molecular Biology. Chapman and Hall.

1997

4. P.K. Gupta.. Elements of Biotechnology. Rastogi and Co. Meerut. 1996.

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MBT-303 ANIMAL CELL SCIENCE AND TECHNOLOGY

UNIT I: Structure and organization of an animal cell, Types of animal cell culture – cell

culture, organ/tissue culture, organotypic culture and histotypic culture, Equipments and

materials needed for animal cell culture technology.

UNIT II: Introduction to the balanced salt solutions and growth medium, Brief

discussion on the chemical, physical and metabolic functions of different constituents of

culture medium, Role of carbon-di-oxide and role of serum and its supplements in

maintaining cells in culture medium, Serum and protein free defined media and their

application

UNIT III: Basic techniques of mammalian cell culture in vitro, primary and established

cell lines, Biology and characterization of the cultured cells, measuring parameters of

growth. Maintenance of cell culture, Cell separation

UNIT IV: Cell transformation, Cell synchronization; Somatic cell genetics, Cell culture

based vaccines, Genetic engineering of mammalian cells in culture, Scaling up of animal

cell culture, Stem cell cultures – embryonic and adult stem cells and their applications

UNIT V: Measurement of viability and cytotoxicity, Apoptosis – characteristic features

and molecular mechanisms, Measurement of cell death, three dimensional culture and

tissue engineering, Applications of animal cell culture technology (heterologous, Primary

culture/CEF culturing, Protein Expression).

PRACTICAL

1. Preparation of tissue culture medium and membrane filtration

2. Preparation of cell suspension from CEF

3. Cell counting and cell viability.

4. Macrophage monolayer from PEC, and measurement of phagocytic activity.

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5. Trypsinization of monolayer and subculturing.

6. Cryopreservation and thawing

7. GFP expression.

BOOKS:

1. Culture of Animal Cells, (3rd Edition), Fl. !an Froshney. Wiley-Liss.

2. Animal Cell Culture - Practical Approach, Ed. John R.W. Masters, OXFORD,

3. Cell Growth and Division: A Practical Approach. Ed. R. Basega, IRL Press.

4. Cell Culture Lab Fax. Eds. M Butler & M. Dawson, Bios Scientific Publications

Ltd..Oxford.

5. Animal Cell Culture Techniques. Ed. Martin Clynes, Springer.

6. Methods in Cell Biology, Vol. 57, Animal Cell Culture Methods. Ed. Jenni P Mather

and David Barnes. Academic Press.

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MBT-304 BIOPROCESS ENGINEERING AND TECHNOLOGY

THEORY

UNIT I: Introduction to Bio process Engineering; Introduction to Biotechnology and

Bioprocess Engineering, bioprocess techniques, biotechnology products. Raw material

used for industrial fermentation and processing, chemical, physical and physiochemical

treatment.

Media for Industrial Fermentation: Introduction, types of media, media formulation;

water, carbon source, energy source, nitrogen source, minerals, buffers, precursors, etc,,

UNIT II: Media for industrial fermentation; Media optimization techniques with

special emphasize on statical techniquies, Placket-Burman design, response surface

methodology, central composite design, etc.

Sterilization; Introduction, media sterilization, the design of batch sterilization process,

the design of continous sterilization process, sterilization of fermentor, sterilization of

feed, sterilization of air and filter design.

UNIT III: Microbial kinetics; Kinetics of microbial growth and death, Monod

equation, yield coefficient, conversion, selectivity.

Overview of Ideal Bioreactors; Batch reactor, Continuous Stirred Tank Reactor

(CSTR), Fed batch and Plug Flow reactors, Cascade reactors, Recycle reactors, Air lift

reactor, fluidized bed bioreactors, Immobilized bed bioreactors, etc,.

Transport phenomena in bioprocess systems; Introduction, oxygen transfer process,

oxygen transfer by aeration and agitation. Determination of oxygen transfer coefficient

by various methods. Factors effecting oxygen transfer coefficient.

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UNIT IV: Downstream processing; Introduction, removal of microbial cells and solid

matter, foam based separation, precipitation, filtration, centrifugation, cell disruptions,

liquid-liquid extraction, chromatography, membrane process, drying and crystallization,

effluent treatment and disposal.

UNIT V: Applications of bioprocess-I; Alcohol (Methanol, Butanol, Ethanol, etc),

Acids (citric acid, Acetic acid ,Gluconic acid etc), Solvents (glycerol, Acetone, Butanol

etc).

Applications of bioprocess-II; Antibiotics (penicillin, Streptomycin, tetracycline)

Aminoacids (Lysine, Glutamic acid), Single Cell Protein, Mineral beneficiation & Oil

recovery.

PRACTICAL

1. Study of enzyme kinetics of invertase

2. Effect of pH on enzyme kinetics

3. Enzyme inhibition

4. Enzyme immobilization experiments with help of different methods.

5. PLACKETT BUKMAN DESIGN for Media design

6. Response surface methodology for media design

7. Sodium sulphite oxidation method for determination of mass transfer coefficient

Dynamic gassing method for determination of mass transfer coefficient

8. Growth kinetics in batch culture.

9. Ethanol production from S. cerevesiae

10. Determination of Enzyme activity for cellulase

11. Pretreatment techniques for lignocellulosic Biomass for ethanol production

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TEXT BOOKS

1) Bailey JE, Ollis DF; Biochemical Engineering fundamentals Year of Publication 1986

2) Blanch HW and Clark DS: Biochemical Engineering Marcel Decker Year of

Publication 1987

3) Introduction to Biochemical Engineering by Dr D G Rao.the Mc Graw Hill companies

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MBT-305 BIOINFORMATICS

UNIT I: Introduction to Bioinformatics -History & Overview and applications, Emerging

areas of Bioinformatics, Primer on: Biology/Genetics/Mol. Genetics, Genomes and their

Importance, genome sequencing, Physical Maps

UNIT II: Biological Databases: Definition, Purpose, Formats, Content, Access - PDB,

Genbank, Swissprot, NRL3D, PIR

DNA Analysis- Strategies for sequencing genomes- Systematic approach, random

approach, EST based approach

Introduction to Whole Genomic analysis, Elements of DNA Sequences, ORF finding,

DNA Motifs and Patterns, Micro-satellite Repeat patterns, Gene Structure (promoter

elements, splicing, termination, etc) Gene Identification: Gene identification methods:

grail, genscan etc

UNIT III : Sequence Alignments and Statistics- Introduction to identity, similarity,

Homology, Sequence Alignment methods, Alignment programs, Pairwise sequence

alignment, Heuristic alignment concept and tools (BLAST- all flavors), mutation matrix,

global vs local alignment, Dot plots, PAM and BLOSUM matrices

Multiple sequence Alignment- algorithms, Phylogenetic studies, dendrograms,

phylograms, cladograms

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UNIT IV: Primer on: Proteins- Types, Functions and applications, Protein Sequence

Analysis -Physico-chemical properties of Amino acids, Protein sequence motifs, Signal

and membrane Proteins, Overview of Protein structure - Principles & characterization,

Secondary structure elements, Domains, Folds, Motifs, Overview of Protein Secondary

Structure Prediction Methods

UNIT V: Introduction to Molecular Modeling concepts and Applications

Overview of Bioinformatics applications -Automated gel reading, Primer Design,

Restriction Endonuclease mapping, Planning of cloning using bioinformatics tools

PRACTICALS:

1. Primer design

2. Pair wise Alignment

3. Multiple sequence Alignment

4. Nucleotide and Amino acid level

5. Construction of phylogenetic tree

6. Gene & Promoter finding by methods of different software’s

TEXT BOOKS:

1. Bioinformatics : Genome and sequence analysis by David W Mount.

2. Bioinformatics : A practical guide to analysis of genes and proteins by Baxevanis,

Andreas D Wiley – Interscience publishers.

REFERENCE BOOKS:

1. Computational Molecular Biology – An Introduction by Peter Clote, Rolf Backofen,

Jhon Wiley & Sons

2. Essential Bioinformatics : by Jin Xiong, Cambridge University Press

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3. Informatics in Proteomics by Srivasthava CRC Publishers.

4. Protein Structure Prediction: Bioinformatics approach by Igor F.Tsigelny

International University Line

5. An introduction to bioinformatics algorithms by Neil C.Jones, Pavel A Pevzner

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M. Sc Biotech 2010 JNTUH

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