Course

1. Preparation of phosphate and acetate buffer. Isolation of Alpha/Beta Amylase from saliva/sweet potato.
2. Standard Maltose Curve, Determination of enzyme activity.
3. Construction of Protein standard curve by Folin’s Lowry method and Determination of specific activity of enzyme.
4. Effect of substrate concentration on Enzyme kinetics and determination of Km and Vmax.
5. Effect of temperature on Enzyme kinetics.
6. Effect of pH on Enzyme kinetics.

LEARNING OBJECTIVES:

Students will begiven hands on exposure to experiments on enzymology which includes preparation of suitable buffer for the isolation of enzymes, velocity of enzymes, protein quantitation, specific activity, kinetics of enzyme and effect of pH and temperature on enzyme kinetics.

COURSE OUTCOMES:

After completing the course, students shall be able to

CO1. Describe and reproduce the fundamentals of calculations and problems in preparation of laboratory solutions and buffers.

CO2. Identify and demonstrate the vigorous methods techniques adopted in the isolation of the enzyme.

CO3. Analyze and conclude the characterization of enzymes by evaluating the velocity, specific activity and kinetic parameters in presence of different factors affecting enzyme kinetics.

• Beedu Sashidhar rao, Vijay Deshpande, “Experimental Biochemistry”, A student companion, 2005 Edition, I K International Pvt. , 9788188237418.
• Jayaraman, Laboratory Manual in Biochemistry; 2011 edition, New Age International Private Limited.
• Lowry, OH, NJ Rosbrough, AL Farr, and RJ Randall.  Biol. Chem. 193:265. 1951