North Maharashtra University, Jalgaon, (With effect from JUNE2017)

 

MB363: Enzymology

 

Unit I: Bioenergetics 20

Role of cofactor in metabolism

Structure and biochemical functions of the following:

1. Nicotinic acid

2. Riboflavin

3. Thiamine

4. Biotin

5. Pantothenic acid

6. Folic acid

 

Unit II: Enzyme Regulation 20

Allosteric enzyme: ATCase

Isoenzyme: LDH

Definition and significance of KM and VMax

Reversible inhibition (Competitive, Uncompetitive and Noncompetitive)

Irreversible inhibition

Covalent modification

Proteolytic modification

 

Unit III: Enzyme Technology 20

Methods of enzyme purification

1. Molecular size: Gel exclusion chromatography

2. Solubility difference: Isoelectric precipitation, salt precipitation

3. Electric charge: Electrophoresis (SDS‐PAGE)

4. Adsorption – Affinity chromatography

Enzyme assay technique (Spectrophotometric assay)

Immobilization: Concept, Methods and Applications

Enzyme engineering : Objectives, Principle and Methodology

 

References

1. Lehninger, A I., (1982), Principles of Biochemistry, Butterworth Publishers, New York.

2. Moat, A., Foster, J., (1988), Microbial Physiology, 2nd Ed., Wiley Interscience

Publications, New York.

3. Palmer T., (1985). Understanding Enzymes. 2nd Ed., Ellis Horwood Ltd., Chichester

4. Price, N. C, Stevens, L, (1989), Fundamentals of Enzymology, 2"d Ed.,Oxford Sci. Publ.,

Oxford

5. Rose, A. H., (1983), Chemical Microbiology, 3rd Ed., Butterworth Publishers.

6. Stamen R. Y., Ingraham, J. L., Wheelis, M. L., Painter, P. R., (1990), General

Microbiology, 5th Ed., MacMillan Edu. Ltd., London

7. Stryer, L., (1988), Biochemistry, W H Freeman and Co., New York.

8. Satyanarayan U., (1999), Biochemistry, Books and Allied (P) Ltd. Calcutta


MB‐361: Molecular Biology

Unit I: Gene Regulation

  • Modes and Mechanism of regulation ‐ Induction and Repression
  • Diauxic growth phenomenon
  • Lac operon ‐Positive and Negative regulation
  • Lac mutants
  • Tryptophan operon


Unit II: Molecular Techniques 

  • Principle and applications of:
    • 1. Blotting techniques ‐ Southern, Northern and Western
    • 2. Autoradiography
    • 3. Gene sequencing ‐ Sanger, Maxam‐Gilbert and Ribotyping
    • 4. Polymerase Chain Reaction (PCR)

  • Gene library and cDNA library
  • Gene mapping ‐ Co‐transformation and interrupted mating experiment


Unit III: r‐DNA Technology

  • Concept and Methodology
  • Restriction endonuclease and DNA ligase
  • Vectors ‐ Plasmid, Cosmid, Phage, YAC
  • Applications of r‐DNA Technology in
    • 1.Health and Medicine: Insulin, Interferon, Hepatitis vaccine
    • 2. Agriculture: BT, Herbicide resistance