NAME: MICHAEL TIMSON

DATE: 1/7/2015
FORM: L6-4
SUBJECT: Biology
TEACHER: Mrs. Mason
TITLE: EFFECTS OF INVERTASE CONCENTRATION ON THE HYDROLYSIS OF
SUCROSE.
AIM: To determine the effects of invertase concentration on the hydrolysis of sucrose.

INTRODUCTION:
Molecules that accelerate, or catalyze a chemical reactions are known as enzymes. In
reactions such as these, the molecules found at the beginning of the process are called substrates.
Enzymes converts these substrates into different molecules, called products. Most metabolic
reactions found in cells, require involution of enzymes catalyst to stimulate these processes at a
faster rate, to sustain life. The type of enzymes synthesized in a cell determines which metabolic
pathways occur in that cell. Enzymes are known to catalyze more than 5,000 biochemical
reaction types. Most enzymes are proteins with the exception of a few being catalytic RNA
molecules. Enzymes' specificity comes from their unique three-dimensional structures. Enzymes
are generally globular proteins, acting alone or in larger complexes. Like all proteins, enzymes
are linear chains of amino acids that fold to produce a three-dimensional structure. The sequence
of the amino acids specifies the structure which in turn determines the catalytic activity of the
enzyme.
Enzymes must bind their substrates before they can catalyze any chemical reaction.
Enzymes are usually very specific as to the substrates they bind and then the chemical reaction
catalyzed. Specificity is achieved by binding pockets with complementary shape, charges and
hydrophilic/hydrophobic characteristics to the substrates. In most biological environments, the
concentration of enzymes is lower than the concentration of substrates. As long as this is true, the

relationship between enzyme concentration and enzyme activity is directly proportional. On a

graph that shows reaction rate versus enzyme concentration, this directly proportional
relationship looks like a straight line with a slope of one. In other words, one additional enzyme
increases the rate by one reaction per unit of time, and one removed enzyme decreases the rate
by one reaction per unit of time.
Sucrose, commonly known as table sugar, is a disaccharide composed of an alphaglucose molecule and a beta-fructose molecule linked by an alpha-1,4-glycosidic bond. When
this bond is cleaved in a hydrolysis reaction, an equimolar mixture of glucose and fructose is
generated. Sucrose can be hydrolyzed in the presence of an enzyme called invertase.

APPARATUS:
Glucose solution, Benedicts Solution, Invertase, Boiling tubes, Water bath,

METHOD
2 cm3 of glucose solution was added to a boiling tube, followed by 2 cm3 of Benedicts
solution. The mixture was heated in a water bath for 5 minutes to obtain a standardized colour
change. 2 cm3 of clear blue Benedicts reagent was added to 2 cm3 of invertase solution. The
mixture was heated in a water bath for 5 minutes. The first step was repeated adding 2% sucrose
solution to 2 cm3 of Benedicts solution. Five clean boiling tubes were labelled and 2 cm3
enzyme concentrations, 0.1%, 0.5%, 1%, 2% and 4%, were each placed in a boiling tube. 2 cm 3
of sucrose solution was then added to each of the boiling tube. The boiling tubes were placed in
the water bath at 35 C and left for 10 minutes to allow then to equilibrate to their surroundings.
1cm3 of the 0.1 percent mixture was added to the boiling tube which containing 1cm 3 of
Benedicts solution. The mixture was then placed in a water bath at 100C. The time the solution
took to form the red precipitate was recorded. The test was repeated using the other mixtures
0.5%, 1%, 2% and 4%.

RESULTS
TABLE SHOWING THE INITIIAL RATE OF HYDROLYSIS OF SUCROSE FOR EACH
CONCENTRATIONS OF INVERTASE
Enzyme

Time taken

Initial Rate of

concentration (%)

T1

T2

Average time taken

Reaction (1/t)

0.1

60.2

64.8

62.5

0.0160

0.5

61.6

61.0

61.3

0.0163

1.0

58.9

58.7

58.8

0.0170

2.0

51.0

54.2

52.6

0.0190

4.0

46.8

48.4

47.6

0.0210

DISCUSSION
From the results obtained in this experiment, it is seen, in tabulated results, that as the
concentration of enzymes increased, the time taken to complete the reaction decreased and thus,
the initial rate of reaction had increased. From the graph, it was seen that between concentrations
of 0.5% to 2%, there was significant increase in the increasing rate of the reaction. That it,
between these concentrations, the increase in the rate of reaction was greatest between these
points. By increasing the concentration of enzyme molecules in the mixture, the number of
particles reacting with the solution are also increased and hence the reaction will take less time to
be completed.
Beyond the 2% concentration, it was observed that the increase in the rate of reaction as a
result of enzyme concentration was decreased as it approached the 4% concentration. Thus, for a
given substrate concentration, the rate of reaction increases with increasing enzyme
concentration up to a point, above which any further increase in enzyme concentration produces

no significant change in reaction rate. This is because there are excess amount of enzyme
molecules with comparison to the substrate. In the increase in concentration of invertase, there
was more active sites present. However the substrate concentration remained the same and thus
the initial rate of reaction remained constant as there were no substrate to bind with the available
active sites.
Within this experiment there were various sources of errors. There were contamination of
reagents to some extent which would have affected the time taken in the results and hence the
rate of hydrolysis of sucrose by invertase. The factor of reaction time was also involved in
stopping the stopwatch and the experiment was conducted in an air conditioning environment
and thus temperature fluctuations occurred. Temperature, being one of the factors affecting initial
rate of reactions of the enzymatic activities, would thus cause a slight depletion in the accuracy
of the results. It is recommended that before conducting the experiment, that all apparatus are
washed thoroughly to ensure that there are no residue of chemicals on the apparatus when
conducting the experiment. Also, the experiment could be conducted using more concentrations,
hence acquiring a more accurate observations pertaining to the effects of the increase of invertase
concentration.

CONCLUSION: By increasing enzyme concentration, the time taken for a reaction to complete
decreased. That is, the initial rate of reaction was increased.