Course Details
Subject {L-T-P / C} : BM3730 : Enzymology and Protein Engineering Laboratory { 0-0-3 / 2}
Subject Nature : Practical
Coordinator : Prof. Nandini Sarkar
Syllabus
Determination of Km and Vmax of an enzymatic reaction, Monitoring effects of pH on enzymatic rate, Monitoring effects of temperature on enzymatic rates, Monitoring effects of substrate concentration on enzymatic rates, ELISA, Monitoring protein stability through equilibrium unfolding method, Studying effects of pH on protein unfolding, Studying effects of temperature on protein unfolding, Monitoring protein folding/unfolding kinetics using stopped flow apparatus, Structural characterization of proteins using extrinsic and intrinsic fluorescence spectroscopy, Determination of secondary structures of a protein using Circular Dichorism (CD) spectroscopy, Monitoring protein-ligand interaction using FRET.
Course Objectives
- To study effects of external parameters like pH, temperature, substrate conc. on enzyme activity
- Protein folding studies using isothermal equilibrium method as well as kinetic studies.
- Structural characterisation of proteins using various biophysical methods
Course Outcomes
After the completion of the course, the students will be able to - <br />CO1 Plan and carry out activity measurements of enzymes and characterize their purity and stability. <br />CO2 Monitor effects of external factors as well as inhibitors on enzyme activity. <br />CO3 Use various biophysical methods for structural and biochemical analysis of proteins. <br />CO4 Monitor and understand effects of various environmental parameters on protein folding/unfolding and stability. <br />CO5 Analyze protein-ligand interactions.
Essential Reading
- G. Walsh, Proteins: Biochemistry and Biotechnology, Wiley Blackwell
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Supplementary Reading
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