DEPARTMENT OF BIOCHEMISTRY - St. Berchmans …sbcollege.ac.in/wp-content/.../2015/03/Biochemistry-complementary.pdf · DEPARTMENT OF BIOCHEMISTRY ... Every student shall undergo at

DEPARTMENT OF BIOCHEMISTRY

Syllabi for Complementary Course for

Model III Industrial Microbiology and Zoology

&

Model III Biotechnology and Botany

Programmes

(with effect from 2015 admissions)

GENERAL OBJECTIVES

The Course intends to provide students with sufficient knowledge of

biomolecular structure to understand the determining properties of biological function

at the level of cells and the body. This will allow students at a later stage:

To understand physiology and physiopathology at the molecular level; the

molecular basis of diagnosis, therapeutics, disease prevention and health

promotion.

Become familiar with and understand the basic structures and functions of cells

in the human body, applying biomedical concepts and terminology.

Apply biochemical analysis and reasoning in order to solve problems related to

physiology and cellular physiopathology.

Learn to use a biochemical approach in the study of cellular functions that will

provide an understanding of future advances in the molecular bases of

physiology, physiopathology, diagnostics, therapeutics, disease prevention,

health promotion and the continuous updating of knowledge.

The Course aims to prepare students:

To acquire and apply the relevant biochemical information in order to solve

potential biomedical problems.

Provide students with basic theoretical and practical knowledge of the principal

methodologies and techniques for investigation of biomolecules; operation,

potential and limitations and selected personal experiences of laboratory work.

Understand the theoretical and practical basis of biochemistry as applied to the

investigation and measurement of cell functions.

Assist students in understanding the scientific method.

Help students develop observation and critical analysis skills: collection,

evaluation and classification of data; deducing conclusions; formulating

hypotheses.

Assist students in developing self-learning and the ability to keep knowledge

and skills up to date; team work and communication.

BOARD OF STUDIES

Members

1. Dr. Jose D Kaipallil

Head, Department of Zoology

S B College

Changanassery

2. Dr. Joe Prasad Mathew

Associate Professor, Department of Zoology

S B College

Changanassery

3. Dr. M S Latha

Professor

School of Biosciences

MG University

Kottayam

4. Dr. Anie Y

Assistant Professor

School of Biosciences

MG University

Kottayam

5. Dr. Valsa A k

Associate professor

Dept. of Microbiology

Sreesankara College

Kaladi

6. Dr. Ajith Joy K

Chairman and Managing Director

DDRC SRL Pvt. Ltd

Ernakulam

7. Mr. V S Sudheesh

Factory head and HR manager

Heart Valve Division

TTK Healthcare Ltd

Thiruvananthapuram

8. Ms. Biby Sara John

Lecturer

Dept. of Biochemistry

S B College

Changanassery

9. Ms. Ramlath Beegum L

Lecturer

Dept. of Biochemistry

S B College

Changanassery

10. Ms. Shema Jacob

Lecturer

Dept. of Biochemistry

S B College

Changanassery

i

Evaluation of the Courses

1. Evaluation

The evaluation of each course shall contain two parts.

i Internal or In-Semester Assessment (ISA)

ii External or End-Semester Assessment (ESA)

Both ISA and ESA shall be carried out using indirect grading. The ISA:ESA ratio shall

be 1:4, for courses.

1.1 In-semester assessment

The components of the internal or in-semester assessment and their marks are as below.

For all courses with practical

Internal assessment of theory courses

There are three components for ISA of theory courses, which include attendance,

assignment/seminar/viva-voce and in-semester examination. All the three components

of the internal assessment are mandatory.

ISA - Components of Theory Marks

Attendance 2

Assignment/Seminar/Viva-Voce 3

In-semester examination (2×2.5 = 5) 5

Total 10

Marks for attendance

% of Attendance Marks

90 and above 2

75 - 89 1

(Decimals shall be rounded off to the next higher whole number)

Internal assessment of practical courses

The internal assessment of practical courses shall be conducted in each semester. The

components for internal assessment are given below.

Internal assessment of practical courses evaluated in each semester

ISA - Components of Practical Marks

Attendance 2

Lab involvement 2

Record* 3

Test (one) 1

Viva-Voce 2

Total 10

ii

*Marks awarded for Record should be related to number of experiments/practicals

recorded.

Marks for attendance

% of Attendance Marks

90 and above 2

75 - 89 1

(Decimals shall be rounded off to the next higher whole number)

1.2 Assignments

Assignments shall be submitted for every course in the first four semesters. At least

one assignment for each course shall be submitted in each semester.

1.3 In-semester examination

Every student shall undergo at least two in-semester examinations as class test as an

internal component for every course.

1.4 To ensure transparency of the evaluation process, the ISA mark awarded to the students

in each course in a semester shall be published on the notice board according to the

schedule in the academic calendar published by the College. There shall not be any

chance for improvement for ISA. The course teacher and the faculty mentor shall

maintain the academic record of each student registered for the course which shall be

forwarded to the office of the Controller of Examinations through the Head of the

Department and a copy should be kept in the office of the Head of the Department for at

least two years for verification.

1.5 A student who has not secured minimum marks in internal examinations can redo the

same before the end semester examination of the semester concerned.

1.6 End-semester assessment

The end-semester examination in theory and practical courses shall be conducted by the

College.

1.7 The end-semester examinations shall be conducted at the end of each semester. There

shall be one end-semester examination of three (3) hours duration in each lecture based

course.

1.8 The question paper should be strictly on the basis of model question paper set by Board

of Studies.

1.9 A question paper may contain short answer type/annotation, short essay type

questions/problems and long essay type questions.

iii

Section Type of Questions

Number of

Questions to be

answered

Marks Total

Marks

A Very short answer type 8 out of 8 1 8

B Short answer type 6 out of 10 2 12

C Short essay solving type 4 out of 6 4 16

D Essay type 2 out of 4 12 24

60

1.10 Photocopies of the answer scripts of the external examination shall be made available to

the students for scrutiny as per the regulations in the examination manual.

1.11 Practical examination shall be conducted in each semester. The duration and frequency

of practical examination shall be decided by the respective Board of Studies.

1.12 Practical examination shall be conducted by one external examiner and one internal

examiner. The question paper setting and evaluation of answer scripts shall be done as

per the directions in the examination manual of the College.

1.13 The marks for end-semester theory and practical examinations are given below

Course Marks

Courses with practical 60

Practical (assessment in each semester) 20

1.14 For all courses (theory and practical) an indirect grading system based on a ten (10)

point scale according to the percentage of marks (ISA + ESA) is used to evaluate the

performance of the student in that course. The percentage shall be rounded

mathematically to the nearest whole number.

Percentage of

Marks Grade Performance Grade Point

90 and above A+ Outstanding 10

80 - 89 A Excellent 9

70 - 79 B Very Good 8

60 - 69 C Good 7

50 - 59 D Satisfactory 6

40 - 49 E Adequate 5

Below 40 F Failure -

1

OUTLINE OF THE COMPLEMENTARY COURSES

Course Code Title of the Course

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Credits ISA ESA Total

Semester I

ADBC101 Biophysical Chemistry 2 36 2 10 60 70

ADBC1P01 Biophysical Chemistry (P) 2 36 1 10 20 30

Semester II

ADBC202 Biomolecules 2 36 2 10 60 70

ADBC2P02 Qualitative Analysis of

Biomolecules (P) 2 36 1 10 20 30

Semester III

ADBC303 Enzymology and Biological

Techniques 3 54 3 10 60 70

ADBC3P03 Enzymology and

Chromatographic Techniques (P) 2 36 1 10 20 30

Semester IV

ADBC404 Metabolism 3 54 3 10 60 70

ADBC4P04 Quantitative Analysis of

Biomolecules (P) 2 36 1 10 20 30

2

3

SEMESTER I

ADBC101: BIOPHYSICAL CHEMISTRY

Total hours of instruction: 36 Credits: 2

Unit I: Water, Acids, Bases and buffers (14 h)

Dissociation of water, ionic product of water, concepts of pH, pOH, simple numerical

problems of pH, determination of pH using indicators, pH meter and theoretical calculations.

Dissociation of weak acids and electrolytes, Bronsted theory of acids and bases, shapes of

titration curve of strong and weak acids and bases. Meaning of Ka and pKa values, Buffers:

buffer action, buffers in biological system, Henderson -Hasselbach equation with derivation,

simple numerical problems involving application of this equation.

Unit II: Solutions (6 h)

Meaning of normality, molarity, molality, percentage solution, mole fractions, simple

numerical problems from the above, Fundamental principles of diffusion and osmosis,

biological importance of osmosis. Isotonic,hypotonic and hypertonic solutions.

Unit III: Colloids (8 h)

Meaning of true solution, colloidal solution, and coarse suspension, distinction between

lyophilic and lyophobic sols, Fundamental study of Donnan equilibrium- application in

biological system, membrane permeability, methods of preparation of colloidal solution,

separation of colloidal solutions, elementary study of charge on colloids, Tyndall effect,

emulsion and emulsifying agents, application of colloidal chemistry.

Unit IV: Isomerism and bonding in biomolecules (8 h)

Classification of isomerism, Intra and Intermolecular interactions in biological system:

Hydrogen bond, Covalent bond, hydrophobic interaction, disulphide bond, Peptide bonds,

glycosidic bond, Phosphodiester linkage, Van der Waal’s forces.

Reference:

A Text Book of Biochemistry (1974) E.S. West, W.R. Todd, H.S. Mason and J.T. van

Bruggen, Oxford and IBH Publishing Co., New Delhi.

Principles of Physical Chemistry (2008) by Puri B R, Sharma L R, Madan S Pathania

Vishal Publishing Co, India ISBN: 8188646008ISBN-13: 9788188646005, 978-

8188646005

4

Biochemistry: A Students survival Guide (2002) Hiram. F. Gilbert Publishers: McGraw-

Hill ISBN 0-07-135657-6

Biochemistry (2012) Reginald H. Garrett and Charles M. Grisham Publisher: 5th Edition,

Cengage Learning, ISBN-13: 978-1133106296, ISBN-10: 1133106293.

A Text Book of Biophysics(2005) R N Roy Publisher: New Central Book Agency(P)

Ltd., Calcutta, India, ISBN: 8173811458

Lehninger Principles of Biochemistry(2004) David L. Nelson, Cox, Fourth Edition ;

Publisher: W. H. Freeman; ISBN-10: 0716743396 ISBN-13: 978-0716743392

5

PRACTICAL

ADBC1P01: BIOPHYSICAL CHEMISTRY

Total hours of instruction: 36 Credits: 1

1. Preparation of solutions:

Percentage solutions

Molar solutions

Normal solutions

Dilution of Stock solutions

2. Bioinstrumentation (Any one to be performed)

a. Use of pH meter

Standardization of pH meter.

Measurements of pH of solutions using pH meters.

b. Colorimetry and Spectrophotometry techniques-Verification of Beer –Lambert’s law

3. Preparation of buffers using the Henderson Hasselbach equation

4. Preparation of Colloidal solutions:

Preparation of Colloidal solution of Prussian blue and Arsenious Sulfide by double

decomposition

Preparation of Colloidal Ferric Hydroxide by Hydrolysis

Reference:

Hawk’s Physiological Chemistry, P.B. Hawk (Author), B. L. Oser (Editor) 14th

Edition Mc Graw Hill Book Company LTD, New York. ISBN-10: 0070478007,

ISBN-13: 978-0070478008

Experimental Biochemistry: A Student Companion (2005), Beedu Sasidhar Rao &

Vijay Deshpande (ed), I.K International Pvt. LTD, New Delhi ISBN 81-88237-41-8.

Biochemical Methods (2008) S. Sadasivam and A. Manickam, New Age International

(P) Limited, ISBN-8122421407, 9788122421408

6

7

SEMESTER II

ADBC202: BIOMOLECULES

Total hours of instruction: 36 Credits: 2

Unit I: Carbohydrates (10 h)

Carbohydrates: Classification, monosaccharides, D and L forms of glyceraldehyde,

Isomerism of carbohydrates, Structure (linear and cyclic structures) of glucose, galactose,

mannose and fructose, epimers and anomers with examples, mutarotation and its explanation

by ring structures, reactions of sugars (due to functional groups - hydroxyl, aldehyde and

ketone), action of acids and alkali on sugars, Reducing actions of sugars., Structure of methyl

α- D glucopyranose, 2- deoxy α- D ribofuranose. Amino sugars, glycosides. Structure and

biological importance of disaccharides-sucrose, lactose, maltose, isomaltose, trehalose and

cellobiose (elucidation of the structures is not included). Structure and important properties of

the following structural polysaccharides (cellulose, chitin, pectin) and storage

polysaccharides (starch, inulin, glycogen). Glycosaminoglycans - heparin, hyaluronic acid.

Unit II: Lipids (8 h)

Lipids: Definition, basic ideas about the biochemical functions of lipids, classification of

lipids with examples, classification of fatty acids, physical and chemical properties of fatty

acids, structure of the following fatty acids- stearic acid, oleic acid, linoleic acid, arachidonic

acid. Structure of triacylglycerol. Saponification number, acid number and iodine number of

fats. Essential and non-essential fatty acids with examples. Compound lipids: storage and

membrane lipids. Structure and functions of phospholipids- phosphatidic acid, lecithin,

cephalin, and phosphatidyl serine, Functions of Sphingolipids. Steroids: Structure and

functions of cholesterol and ergosterol.

Unit III: Proteins (8 h)

Amino acids: Definition, stereoisomerism, structure of 20 standard amino acids - single letter

abbreviations of amino acids, classification of amino acids based on charge and polarity,

essential and non essential amino acids, ionization of amino acids. Non-standard amino acids,

Amino acid derivatives of biological significance, Peptides: Formation of peptide

bond. Classification of proteins. General properties of proteins. Denaturation of proteins and

precipitation reactions of proteins. Elementary study of primary, secondary, tertiary and

8

quaternary structure of proteins, forces stabilizing the structure of protein. Outline of protein

sequencing (only basic principles of the methods employed).

Unit IV: Nucleic acids (6 h)

Nature of nucleic acids, Structure of purines and pyrimidines, nucleosides, nucleotides,

Stability and formation of phosphodiester linkages, Effect of acids, alkali and nucleases on

DNA and RNA, Structure of Nucleic acids- Watson-Crick DNA double helix structure, Brief

study of: circular DNA, super coiling, helix to random coil transition, denaturation of nucleic

acids- hyperchromic effect, Tm-values and their significance, Types of RNA and DNA,

Unusual bases in nucleic acids. DNA sequencing: Sanger and Dideoxy methods.

Unit V: Vitamins and Minerals (4 h)

Vitamins: Definition, classification- fat-soluble and water-soluble: sources, chemical nature

(without structure), and functions of vitamins. Minerals: requirements, macro and micro

minerals (source and functions).

Reference:

Lehninger Principles of Biochemistry(2004) David L. Nelson, Cox, Fourth Edition ;

Publisher: W. H. Freeman; ISBN-10: 0716743396 ISBN-13: 978-0716743392

A Text Book of Biochemistry (1974) E.S. West, W.R. Todd, H.S. Mason and J.T. van

Bruggen, Oxford and IBH Publishing Co., New Delhi.

Biochemistry (2004) Donald Voet, Judith G. Voet Publisher: John Wiley & Sons Inc

ISBN: 047119350X ISBN-13: 9780471193500, 978-0471193500

Principles Of Biochemistry (1995), Geoffrey L Zubay, William W Parson, Dennis E

Vance Publisher: Mc Graw-Hill Book Company, Koga ISBN:0697142752

Principles Of Biochemistry (2006); Robert Horton H, Laurence A Moran, Gray

Scrimgeour K Publisher: Pearsarson ISBN: 0131977369, ISBN-13:9780131977365, 978-

0131977365

Biochemistry (2007) Jeremy M. Berg John L. tymoczko Lubert Stryer Publisher: 6th

Edition B.I. publications Pvt. Ltd ISBN:071676766X ISBN-13: 9780716767664, 978-

716767664

Biochemistry (2008); Rastogi, Publisher: Mcgraw Hill ISBN:0070527954 ISBN-13:

9780070527959, 978-0070527959

Fundamentals of Biochemistry (2008) J. L. Jain, Sunjay Jain and Nitin Jain Publishers: S.

Chand & Co Ltd ISBN: 81-219-2453-7.

9

Biochemistry (2012) Reginald H. Garrett and Charles M. Grisham 5th Edition Publisher:

CengageLearning,ISBN-13: 978-1133106296 ISBN-10: 1133106293

Harpers Illustrated Biochemistry(2012), Robert Murray, David Bender, Kathleen M.

Botham, Peter J. Kennelly, Victor Rodwell and P. Anthony Weil,29th Edition (LANGE

Basic Science); Publisher: McGraw-Hill Medical; ISBN 10: 007176576X;ISBN-

13: 9780071765763

10

11

PRACTICAL

ADBC2P02: QUALITATIVE ANALYSIS OF BIOMOLECULES

Total hours of instruction: 36 Credits: 1

1. Reactions of Carbohydrates, Amino acids, Proteins, Lipids and Non protein

nitrogenous compounds

A. Carbohydrates: (Glucose, fructose, Galactose, Xylose, Maltose, Lactose, Sucrose,

Starch, dextrin maybe given for analysis).

Molisch’s test, Iodine test, Test for reducing sugars (Fehling’s test, Benedict’s test,

Barfoed’s test), Seliwanoff’s test, Bial’s test, Mucic acid test, Acid hydrolysis of

Sucrose, Osazone test.

B. Amino acids: (tyrosine, tryptophan, cysteine, cystine, methionine, arginine, proline,

histidine may be given for analysis)

Ninhydrin test, Xanthoproteic test, Isatin test, Pauly’sdiazo test, sakaguchi test,

Ehrlich’s test, Sodium nitroprusside test, Millon’s test, Sullivan’s test.

C. Proteins: (Casein, Albumin, Gelatin, peptone may be given for analysis).

Biuret test, Ammonium sulfate precipitation test, Sulphosalicylic acid test, Heat

coagulation test, test for inorganic phosphate.

D. Lipids: Fats- tristearin, Fatty acids- palmitic acid, stearic acid, oleic acid, Glycerol,

Steroids, cholesterol

Solubility in Organic solvents, saponification test, Acrolein test, Test for unsaturation:

with bromine water or dilute potassium permanganate or Hubl’s iodine test, salkowski

test, Zak’s test

E. Non protein nitrogenous compounds: (Urea, Uric acid, creatinine)

Urease test, Phosphotungstic acid test and Jaffe’s test

2. Identification of Monosaccharide, Disaccharide, polysaccharide from a samplefollowing a

systematic scheme of analysis (only single component of above mentioned

carbohydrates).

3. Identification of amino acids and proteins following a systematic scheme for analysis

(single components only need be given)

4. Identification of lipids following a systematic scheme for analysis (single components

only need be given)

12

5. Identification of NPN following a systematic scheme for analysis (single components

only need be given)

Reference:

Hawk’s Physiological Chemistry, P.B. Hawk (Author), B.L. Oser (Editor) 14th Edition

Mc Graw Hill Book Company LTD, New York. ISBN-10: 0070478007, ISBN-13: 978-

0070478008

Experimental Biochemistry: A Student Companion (2005), Beedu Sasidhar Rao& Vijay

Deshpande (ed), I.K International Pvt. LTD, New Delhi ISBN 81-88237-41-8.

Introductory Practical biochemistry, S. K. Sawhney & Randhir Singh (eds) Narosa

Publishing House, New Delhi, ISBN 81-7319-302-9 p 15- 23, 33- 35, 50 -57.

Practical Biochemistry, R.C. Gupta & S. Bhargava (eds) CBS Publishers and Distributors,

New Delhi, ISBN 81-239-0124-0 p 9 – 27.

Biochemical Methods (2008) S. Sadasivam and A. Manickam, New Age International (P)

Limited, ISBN-8122421407, 9788122421408

13

SEMESTER III

ADBC303: ENZYMOLOGY AND BIOLOGICAL TECHNIQUES

Total hours of instruction: 54 Credits: 3

Unit I: Introduction to enzymes (14h)

History of enzymology, holoenzyme, apoenzyme, and prosthetic group. Activation energy.

Interaction between enzyme and substrate- lock and key model, induced fit model. Features

of active site.Enzyme specificity and types with example. Enzyme Commission system of

classification and nomenclature of enzymes: six major classes of enzymes with one example

each. Ribozymes, Abzymes. Coenzymes and their functions - NAD, NADP+, FAD, FMN,

lipoic acid, TPP, pyridoxal phosphate, biotin and cyanocobalamin. Measurement and

expression of enzyme activity, Definition of IU, katals, enzyme turnover number and specific

activity.

Unit II: Enzyme kinetics (14 h)

Elementary study of the following factors affecting velocity of enzyme catalysed reactions-

effect of substrate concentration, enzyme concentration, temperature and pH; Km and its

significance, Michaelis Menton equation (without derivation), Line weaver- Burk

plot, Explanation of competitive and non competitive type of inhibition, their explanation on

the basis of double reciprocal plot. Brief study of allosteric regulation with an example. Brief

study of the activation of zymogen form of enzymes, covalent modification of enzymes.

Isoenzymes- Lactate dehydrogenase and creatine phosphokinase. Elementary study of

isolation of enzymes and the criteria of purity.

Unit III: Industrial applications of enzymes (8 h)

Immobilization of enzymes, methods of immobilization, Industrial uses of enzymes:

production of glucose from starch, cellulose and dextrans, use of lactase in diary industry,

production of glucose fructose syrup from sucrose, use of proteases in food, leather and

detergent industry. Diagnostic and therapeutic enzymes (brief study of name of enzyme and

role in diagnosis and therapy)

Unit IV: Biological Techniques (18h)

Chromatography - types of chromatography- adsorption chromatography, partition

chromatography, ion exchange chromatography, gel permeation chromatography and affinity

chromatography.

14

Electrophoresis - Principle, procedure and application of Agarose gel electrophoresis,

Polyacrylamide gel electrophoresis, Isoelectric focussing, blotting techniques.

Colorimetry and Spectrophotometry: - Principles of colorimetry and spectrophotometry.

Differential Centrifugation

ELISA, Chemiluminescence, Immunofluorescence.

Reference

Lehninger Principles of Biochemistry(2004) David L. Nelson, Cox, Fourth Edition;

Publisher: W. H. Freeman; ISBN-10: 0716743396 ISBN-13: 978-0716743392

A Text Book of Biochemistry (1974); E.S. West, W.R. Todd, H.S. Mason and J.T. van

Bruggen, Oxford and IBH Publishing Co., New Delhi.

Biochemistry (2004) Donald Voet, Judith G. Voet Publisher: John Wiley & Sons

Inc ISBN: 047119350X ISBN-13: 9780471193500, 978-0471193500

Principles of Biochemistry (1995) Geoffrey L Zubay, William W Parson, Dennis E

Vance Publisher: Mc Graw-Hill Book Company Koga ISBN:0697142752 ISBN-

13: 9780697142757, 978-0697142757

Principles of Biochemistry, (2006) Robert Horton H , Laurence A

Moran, GrayScrimgeourK Publisher: Pearsarson ISBN: 0131977369, ISBN-

13:9780131977365, 978-0131977365

Biochemistry (2007) Jeremy M. berg John L. Tymoczko Lubert Stryer 6th

Edition Publisher: B.I. Publications Pvt. Ltd ISBN:071676766X ISBN-

13: 9780716767664, 978-716767664

Biochemistry (2008) Rastogi Publisher: McgrawHill ISBN:0070527954 ISBN-

13: 9780070527959, 978-0070527959

Fundamentals of Enzymology: The Cell and Molecular Biology of Catalytic Proteins

(2000) Nicholas C. Price, Lewis Stevens, and Lewis Stevens Publisher: Oxford University

Press, USAISBN: 019850229X ISBN-13: 9780198502296, 978-0198502296

Enzyme Mechanism (2007) P.K. Shivraj Kumar Publisher: RBSA Publishers

ISBN: 8176114235 ISBN-13: 9788176114233, 978-8176114233

Enzymes: Biochemistry, Biotechnology, Clinical Chemistry (2007) Trevor Palmer, Philip

Bonner, Second Edition, Publisher: Horwood Publishing Limited ISBN: 1904275273 ISBN-

13: 9781904275275, 978-1904275275

Fundamentals of Biochemistry (2008) J. L. Jain, Sunjay Jain and Nitin Jain Publishers: S.

Chand & Co Ltd ISBN: 81-219-2453-7.

15

Introduction to Biophysics (2008) Pranabkumar Banerjee Publishers: S. Chand &

Company Ltd ISBN: 81-219-3016-2 p 32.

The Tools of Biochemistry (1977) Cooper T.G. John Wiley and Sons, N.Y. USA.

16

17

ADBC3P03: ENZYMOLOGY AND CHROMATOGRAPHIC

TECHNIQUES

Total hours of instruction: 36 Credits: 1

Objective: The objective here is to make the students understand the basic steps involved in

extraction and determination of enzymatic activities & Calculation enzymatic activities from

experimental data

1. Extraction of enzymes: (Minimum 3 experiments to be done)

Acid phosphatase from Fresh Potato (Solanumtuberosum)

β- amylase from Sweet potato (Ipomoea batates)

Catalase from Bovine /Porcine liver

Urease from Jack bean (Canavaliaensiformis)

Phytase from Seeds

2. Enzyme Assay: (Minimum 2 experiments to be done, enzymes extracted from above

experiment can be used)

Acid phosphatase

β- amylase

Catalase

Urease from Jack bean

Phytase

3. Biochemical separation techniques- Chromatographic techniques (Any one to be

performed)

Separation of amino acids and simple sugars by Paper chromatography (Descending or

ascending)

Separation of amino acids and lipids by Thin Layer chromatography

Separation of Plant pigments by Column chromatography

References

Experimental Biochemistry: A Student Companion (2005), Beedu Sasidhar Rao

(author)& Vijay Deshpande (ed), I.K International Pvt. LTD, New Delhi ISBN 81-88237-

41-8.

18

Introductory Practical biochemistry, S. K. Sawhney & Randhir Singh (eds) Narosa

Publishing House, New Delhi, ISBN 81-7319-302-9, p 110 - 155

Standard Methods of Biochemical Analysis, S. K. Thimmaiah (ed), Kalyani Publishers,

Ludhiana ISBN 81-7663-067-5, p 184 – 255.

Biochemical Methods (2008) S. Sadasivam and A. Manickam New Age International (P)

Limited, ISBN-8122421407, 9788122421408

19

SEMESTER IV

ADBC404: METABOLISM

Total hours of instruction: 54 Credits: 3

Unit I: Carbohydrate metabolism (16 h)

Digestion of carbohydrates and absorption of sugars, Reactions of glycolytic sequences with

the names of enzymes and intermediates (with structures), Fate of pyruvate in alcoholic

fermentation, Substrate level phosphorylation. Glycogenesis and glycogenolysis, Role of

cyclic AMP and hormones in glycogen metabolism, Gluconeogenesis and pentose phosphate

pathway (with structures of intermediates). Decarboxylation of pyruvate, Reactions of citric

acid cycle (with structures of intermediates), Calculation of energy yield (as ATP) of aerobic

and anaerobic oxidation of carbohydrates, the mitochondria, arrangement of electron carriers

in the electron transport chain: Oxidative phosphorylation, site of ATP formation in the

electron transport chain, Chemiosmotic hypothesis.

Unit II: Lipid metabolism (14 h)

Outline study of lipid digestion and absorption. Outline study of β-oxidation scheme (with

structures). ATP yield in β-oxidation, Cytoplasmic biosynthesis of fatty acid, elongation and

desaturation, Physiological functions of phospholipids, Outline study of cholesterol synthesis

(without structure).

Unit III: Protein metabolism (10 h)

Proteolytic enzymes of the gastrointestinal tract and their activation (from zymogen forms),

Digestion of proteins, Absorption of aminoacids from the intestine, Decarboxylation,

deamination and transamination of aminoacids (without molecular mechanisms), Urea cycle,

Fate of carbon skeleton: entry into glucogenic and ketogenic pathways. N2 fixation (detailed

study expected).

Unit IV: Nucleotide metabolism (6 h)

Metabolism of nucleotide: Biosynthesis of purine and pyrimidine nucleotides -de novo and

salvage pathway (no structure required), end products of purine and pyrimidine metabolism.

Unit V: Photosynthesis (8 h)

Light reactions: cyclic and non-cyclic electron transport and photophosphorylation. Dark

reactions: the path of carbon- C3 & C4 Pathways (structure not needed), glyoxylate cycle and

its significance.

20

Reference:

Lehninger Principles of Biochemistry(2004) David L. Nelson, Cox, Publisher: W. H.

Freeman; Fourth Edition ; ISBN-10: 0716743396 ISBN-13: 978-0716743392

A Text Book of Biochemistry (1974), E.S. West, W.R. Todd, H.S. Mason and J.T.

van Bruggen, , Oxford and IBH Publishing Co., New Delhi,

Biochemistry (2004) Donald Voet, Judith G. Voet Publisher: John Wiley & Sons Inc

ISBN: 047119350X ISBN-13: 9780471193500, 978-0471193500

Principles Of Biochemistry (1995) Geoffrey L Zubay, William W Parson, Dennis E

Vance Publisher: Mc Graw-Hill Book Company, Koga ISBN:0697142752

Principles Of Biochemistry (2006) Robert Horton H, Laurence A Moran, Gray

Scrimgeour K Publisher: Pearson ISBN: 0131977369, ISBN-13:9780131977365,

978-0131977365

Biochemistry (2007) Jeremy M. Berg John L. Tymoczko Lubert Stryer 6th Edition

Publisher: B.I .publications Pvt. Ltd ISBN:071676766X ISBN-13: 9780716767664,

978-716767664

Biochemistry (2008) Rastogi Publisher: Mc Graw Hill ISBN:0070527954 ISBN-13:

9780070527959, 978-0070527959

Harpers Illustrated Biochemistry(2012),Robert Murray, David Bender, Kathleen M.

Botham, Peter J. Kennelly, Victor Rodwell and P. Anthony Weil29th Edition

(LANGE Basic Science)Publisher: McGraw-Hill Medical;ISBN-

10: 007176576X;ISBN-13: 9780071765763

21

ADBC4P04: QUANTITATIVE ANALYSIS OF BIOMOLECULES

Total hours of instruction: 36 Credits: 1

A. Estimation of Carbohydrates: (Colorimetric)

1. Quantitation of total sugars by anthrone method

2. Determination of reducing sugars by Nelson and Somogyi’s method

3. Determination of fructose by Roe’s resorcinol method

B. Separation and Estimation of Lipids: (Colorimetric)

1. Estimation of Cholesterol by Zak’s method

2. Determination of acid value of fats

3. Determination of peroxide value of oils

C. Estimation of Proteins and Amino acids: (Colorimetric)

1. Estimation of protein by Lowry’s method

2. Determination of protein by Biuret method

D. Estimation of Nucleic acids: (Colorimetric)

1. Estimation of DNA by Diphenylamine method

2. Determination of RNA by orcinol method

Reference:

Experimental Biochemistry: A Student Companion (2005), Beedu Sasidhar Rao

(author) & Vijay Deshpande (ed), I.K International Pvt. LTD, New Delhi ISBN 81-

88237-41-8.

Introductory Practical biochemistry, S. K. Sawhney & Randhir Singh (eds) Narosa

Publishing House, New Delhi, ISBN 81-7319-302-9, p 15 - 109

Standard Methods of Biochemical Analysis, S. K. Thimmaiah (ed), Kalyani

Publishers, Ludhiana ISBN 81-7663-067-5, p 49- 181, p 269- 285.

Biochemical Methods (2008) S. Sadasivam and A. Manickam New Age International

(P) Limited, ISBN-8122421407, 9788122421408.