Catalase

enzyme
observe how it is affected by concentration of
enzyme and substrate, temperature, pH, and
enzyme inhibitor

biological catalysts

Enzymes that carry out the thousands of

reactions that occur in living cells

prosthetic group

- generally large proteins

- made up of several hundred amino acids
- contain a non-protein group- prosthetic group
-important to catalytic reaction

substrate

In an enzyme-catalyzed reaction, the substance

to be acted upon binds to the active site of the
enzyme.

enzyme-substrate complex
holds enzyme and substrate together by

1. Enzyme-substrate complex holds enzyme

and substrate together with hydrophobic
bonds, hydrogen bonds, and ionic bonds
2. The enzyme then converts the substrate to
the reaction products in a process that often
requires several steps, and may involve the
breaking and formation of covalent bonds
3. Finally, the products are released into
solution and the enzyme is ready to form
another enzyme-substrate complex.

true of any enzyme

catalyst is not used up as it carries out the

reaction but is recycled

Each enzyme is specific for a certain reaction

because

its amino acid sequence is unique and causes it

to have a unique three-dimensional structure

active site

The business end of the enzyme molecule has

a specific shape so that only one of the
thousands of compounds present in the cell can
interact with it
- prosthetic group on the enzyme, it will
form part of the active site

prosthetic group on the enzyme

Any substance that blocks or changes the shape
of the active site will
denatured
determining the enzymes shape

interfere with the activity and efficiency of the

enzyme

-if large enough change; enzyme no

longer acts = denatured
Salt concentration, pH, Temp

Salt concentration
Salt is low or zero
1. Charged amino acid side chains of enzyme
stick together
2. Denature and form an inactive precipitate
Salt is high
1. Normal interaction of charged groups will
be blocked
2. New interactions occur
3. And again the enzyme precipitates
intermediate salt concentration
Blood salt concentration is?

(0.9%)
Is optimum for most enzymes.

pH
Enzyme amino acid side chains contain groups
such as
pH is lowered

pH is raised

COOH, and NH2 that readily gain or lose H+

ions
1. enzyme gain H+ ions
2. Enough side chains will be affected so the
enzyme shape is disrupted
1. enzyme will lose H+
2. lose its active shape

enzymes have an optimum pH in the

neutral pH range and are denatured at either

extremely high or low pH but in stomach those
will be appropriately low pH at optimum level

Temperature

All chemical reactions speed up as the

temperature is raised; same for enzymes
temperature increases; more kinetic energy to
undergo reaction

Temp increased past optimum level will

disrupt enzyme molecule
Many proteins are denatured by temperatures of
____ but still active at ____C and even
withstand______

40-50 70-80 being boiled

Small Molecules

Most small molecules will interact with

enzymes other than substrates

activator

molecule increases the rate of the reaction

inhibitor

decreases the rate of the reaction; denatures

cell can use these molecules to regulate

how fast the enzyme acts

Some inhibitors act by

reducing the -S-S- bridges that stabilize the

enzymes structure

Many inhibitors act by

reacting with side chains in or near the active

site to change or block it

enzyme inhibitors interfere with the active site

examples

potassium cyanide
curare

What does catalase do?

accelerates the breakdown of hydrogen peroxide

(a common end product of oxidative
metabolism) into water and oxygen

Formula for Catalase

catalase-mediated reaction important because

Catalase is found in

Exercise 6.1 Extraction of Catalase

enzyme activity

2 H2O2 + catalase

2 H2O + O2 + catalase

prevents the accumulation of hydrogen peroxide,

a strong oxidizing agent which can disrupt cell
animal tissue
plant tissues
-abundant in plant storage organs
-such as potato tubers
-turnips
-fleshy parts of fruits
1. will isolate catalase from potato tubers
2. measure its rate of activity under
different condition
3. glass fiber filter will be immersed in the
enzyme solution
4. Then placed in the hydrogen peroxide
substrate
5. oxygen produced from the subsequent
reaction will become trapped in the disk
and give it buoyancy
time measured from disc touching solution to
time it reaches the surface will be rate of the
enzyme activity

Exercise 6.2- Effect of Enzyme

Concentration
First demonstrate enzyme follows chemical
principles by

determining the effect of enzyme concentration

on the rate of activity, while using a substrate
concentration which is in excess

Exercise 6.3 Effect of Substrate Concent

Exercise 6.4 Effect of Temperature

Exercise 6.5 Effect of pH
Exercise 6.6- Effect of Enzyme Inhibitor
To determine the effect of substrate
concentration on enzyme activity prepare 40 mL
of the appropriate H2O2 solution

Hydroxylamine attaches to an iron atom (a part

of the catalase molecule) and thereby interferes
with the formation of the enzyme-substrate
complex.