7 - Optimum Conditions for Enzymatic Activity of -Amylase...

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Optimum Conditions for Enzymatic Activity of α -Amylase from Human Saliva Objectives • A. Enzymatic Degradation of Starch with α -amylase. • B. Measurement of starch concentration • C. Dependence of α -amylase Catalytic Activity on pH Things You Should Know After Completing Exercise 7 Optimum Conditions for Enzymatic Activity of α -Amylase How to make a parallel or serial dilutions of a stock starch solution. •H o w t o u s e I 2 +KI solution to test for the presence of starch in a solution. How to use a spectrophotometer to measure the starch concentration of a solution containing a starch and I 2 +KI solution. How to determine the saliva dilution that cause the rate of decrease of A 620 closest to 0.3/min. • How to plot A 620 values vs. time on a linear graph, calculate the slopes of the lines and identify the pH that provides the highest enzymatic activity. Enzymes • A protein that acts as a catalyst in biochemical reactions by forming an enzyme-substrate complex. • Enzymes accelerate the rate of a reaction by lowering the energy of activation needed to start a spontaneous reaction. Enzymes Catalyze Reactions (speed up reaction rate 10 5 -10 17 times) SUCROSE GLUCOSE & FRUCTOSE ± ! In the absence of an enzyme, even a spontaneous reaction occurs very slowly. Sucrose Sucrose Sucrose Glucose & Fructose 100 years 5 minutes Sucrase Enzyme ± ! ± !
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Some Characteristics of Enzymes • Substrate specific • Temperature (50 C+ lose tertiary structure) • pH (many denatured by pH extremes) • Salt composition of environment (affects ionic and hydrogen bonding) • Often associated w/ cofactors and coenzymes Other Factors Affecting Rate of Reaction • Concentration of: • (1) Ions in the buffer
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This note was uploaded on 02/22/2010 for the course BIO 50100 taught by Professor Brand during the Fall '09 term at University of Texas at Austin.

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7 - Optimum Conditions for Enzymatic Activity of -Amylase...

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