Factors Affecting Enzyme Action

Rate: measured by the amount of substrate changed, or amount of product formed, during a period of time. Determine by measuring the slope of the tangent to the curve in the initial stage of the reaction. The steeper the slope, the greater the rate. If activity is measure over a period of time, the rate of reaction usually falls.

Optimal conditions favour the most active conformation for the enzyme molecule. Conditions that affect rate of enzymes:
pH

Temperature
[enzyme] [substrate]

Co-factors
Enzyme inhibitors

Maximum activity at optimum pH When pH value is altered above or below this value, the rate of enzyme activity diminishes. As pH decreases, acidity increases and the concentration of H+ ions increases. This increases the number of positive charges in the medium.

Changes in pH alter the ionic charge of the acidic and basic groups and therefore disrupt the ionic bonding that helps maintain the specific shape of the enzyme. The pH change leads to the alteration of the shape and the active site. In extremes conditions the enzyme will denature.

Rate of reaction

10

12 14

pH

Optimum pH
6

little activity at low temperature rate increases with temperature most active at optimum temperatures (e.g. 37C in humans) activity lost at high temperatures

Optimum temperature

Reaction rate

Temperature / oC
9

Low temperature :
enzyme inactive rate low

Increasing temperature :

increase K.E. / molecular motion of E & S increase frequency of effective collisions more E-S complex formed rate increases
10

Optimum temperature :
max activity

Above optimum temperature :

H bonds broken 2o & 3o structure disrupted active site configuration altered enzyme denatured rate decreases

11

increasing [E] increases rate of reaction ([S] constant) rate of change decreases when less substrate available max activity reached when limited by [S]

12

Maximum activity (Vmax)

Rate of reaction

Enzyme concentration
13

increasing [S] increases the rate of reaction ([E] constant) rate of change decreases as less active site available max activity reached when all active sites saturated

14

Maximum activity (Vmax)

Rate of reaction

Substrate concentration
15

Sucrase has an optimum temperature of 37C and an optimum pH of 6.2. Determine the effect of the following on its rate of reaction (1) no change (2) increase (3) decrease A. Increasing the concentration of sucrose B. Changing the pH to 4 C. Running the reaction at 70C

Sucrase has an optimum temperature of 37C and an optimum pH of 6.2. Determine the effect of the following on its rate of reaction (1) no change (2) increase (3) decrease A. 2, 1 Increasing the concentration of sucrose B. 3 Changing the pH to 4 C. 3 Running the reaction at 70C

Protease, which is an enzyme used in biological washing powders, is an enzyme that conducts proteolysis, that is, begins protein catabolism by hydrolysis of the peptide bonds that link amino acids together in the polypeptide chain forming the protein. a. How would a protease remove a blood stain on clothes? b. Most biological washing powders are recommended for use at low washing temperatures. Why is this? c. Washing powder manufacturers have produced proteases which can work at higher temp. than 40C. Why is this useful?

a. Haemoglobin colours blood stains. Protein-digesting enzymes hydrolyse haemoglobin to amino acids, which are colourless. They are also soluble, so will wash away in water.
b. Many protein-digesting enzymes have an optimum temp. of around 40C.

c. Other components of washing powders, such as the oil-removing detergents, work best at high temperatures.