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Enzyme Kinetics: Effect of Temperature and pH

An enzyme look more like a tangled chain of amino acids that strung up together. If it

zoomed even further you’d see tons of tons of atoms. And without this atoms s staying in the right

spot the substrate would not be able to fit in the active site of the enzyme. So in what conditions

do the atoms actually stay put and keep their shape and in what conditions do they fail to do so?

There are three conditions that affect this the most: the effect of temperature, pH and the substrate


Effect of Temperature on Enzyme Activity

When enzyme and substrate molecules are mixed they randomly collide forming enzyme

substrate complexes. Like most chemical reactions, as temperature increases so does the kinetic

energy of the molecule and therefore also the rate at which these collisions occur. Optimum

temperature is the temperature at which the maximum rate of reaction is achieved. A ten-degree

Centigrade rise in temperature will increase the activity of most enzymes by 50 to 100%.

Variations in reaction temperature as small as 1 or 2 degrees may introduce changes of 10 to 20%

in the results. If the temperature continues to increase beyond the optimum temperature, the

enzymes begin to lose their three-dimensional structure and becomes denature, therefore reducing

the rate of reaction.

As shown in the figure, the reaction rate increases

with temperature to a maximum level, then

abruptly declines with further increase of

temperature. Because most animal enzymes rapidly become denatured at temperatures above

40°C, most enzyme determinations are carried out somewhat below that temperature.

Over a period of time, enzymes will be deactivated at even moderate temperatures. Storage

of enzymes at 5°C or below is generally the most suitable. Some enzymes lose their activity when


Effect of pH on Enzyme Activity

Enzymes are affected by changes in pH. The most favorable pH value - the point where the enzyme

is most active - is known as the optimum pH. Extremely high or low pH values generally result in

complete loss of activity for most enzymes. This is graphically illustrated in the graph below.

Increasing pH values are as a result of adding hydroxide ions or OH-, decreasing pH is a

result of adding hydrogen ions H+. Either of these can potentially alter the charges on the amino

acids that make up the enzyme and this can lead to the enzymes active site changing shape because

the bonding changes for example and that could denature the enzyme.
The optimum pH value will vary greatly from one enzyme to another, as Table I shows:

Table I: pH for Optimum Activity

Enzyme pH Optimum

Lipase (pancreas) 8.0

Lipase (stomach) 4.0 - 5.0

Lipase (castor oil) 4.7

Pepsin 1.5 - 1.6

Trypsin 7.8 - 8.7

Urease 7.0

Invertase 4.5

Maltase 6.1 - 6.8

Amylase (pancreas) 6.7 - 7.0

Amylase (malt) 4.6 - 5.2

Catalase 7.0

The Effect of Substrate Concentration on Enzyme Activity

When the substrate concentration change, it is assumed that the enzyme concentration remains

constant. Initially, increasing substrate concentration from zero will increase the rate of reaction

as the substrate randomly collide with the enzymes and the enzymes catalyze the chemical

reaction. However, there comes a point where all of the enzyme active sites are filled and the
enzymes are already working as quickly as they can to catalyze the reaction, this is where the

maximum rate of reaction is reached and any further addition of substrate beyond this will not

increase the rate of reaction anymore as shown in the figure below.

Introduction to Enzymes. (n.d.). Retrieved June 19, 2017, from http://www.worthington- Effect of pH on Enzyme Activity

Introduction to Enzymes. (n.d.). Retrieved June 19, 2017, from http://www.worthington- Effect of Temperature on Enzyme Activity

Honey & Acetic Acid Research Proposal | Fixation ... (n.d.). Retrieved June 19, 2017, from




B. (2014, October 16). Retrieved June 19, 2017, from