The Rate Of Reaction Of Succinate Dehydrogenase

1537 words - 7 pages

The rate of reaction of Succinate dehydrogenase

Introduction:

Enzymes are protein molecules that function as biological catalysts
that can help break larger molecules into smaller molecules while
remaining unchanged themselves. They speed up the chemical reactions
by lowering the energy of activation barrier, are specific to one
molecule. The enzyme’s specificity arises from its active site, an
area with a shape corresponding to the molecule with which it reacts
(the substrate). The shape of the enzyme where the chemical binds only
allows the binding of that particular chemical, or inhibitor
substrates that are structually similar to the substrate, competing
for the active site. The enzyme ...view middle of the document...

3 test tubes

Distilled water

0.2M Na2HPO4 with 100ml distilled water (solution 1)

0.2M NaH2PO4 with 100ml distilled water (solution 2)

0.1M succinate with 100ml distilled water

0.0003 M DPIP with 100ml distilled water

BUFFERS

pH 5 buffer – 1ml solution 1 to 49ml of solution 2, mix – add 50 ml
distilled water

pH 7.3 buffer – 75ml solution 1 to 25ml solution 2, mix – add 100ml
distilled water

pH 9 buffer - 10ml solution 1 to 10ml distilled water

Stopwatch

Enzyme preparation: (Wright, 2005)

Chicken hearts were prepared according to notes in Lab (Wright 2005).

This liquid formed the enzyme.

A rack containing 3 test tubes were arranged containing:

tube 1 – 5.8 ml pH5 buffer

1 ml 0.1M succinate

.2 ml enzyme

tube 2. - 5.8 ml pH7.3 buffer

1 ml 0.1M succinate

.2 ml enzyme

tube 3. - 5.8 ml pH9 buffer

1 ml 0.1M succinate

.2 ml enzyme

1 ml of 0.0003M DPIP was added to each tube, noting the time the
addition was made. A dropper full of cooking oil was added to cover
surface of solution. The time taken for the blue colour to disappear
or become very faint, was recorded.

This procedure was repeated for tubes 2 and 3 (Wright, 2005).

Part 2. Effect of temperature on enzyme activity

3 test tubes were arranged as in part one, but using the pH buffer 7.3
in each tube.

Tube 1 was inserted into iced water, measured to be 4 degrees Celsius.

Tube 2 was left at room temperature, measured to be 17 degrees Celsius

Tube 3 was inserted into hot tap water measuring 41 degrees Celsius.

Reaction times were recorded.

Part 3. Activity of succinate dehydrogenase with different
substances

A set of 5 test tubes were arranged containing solutions succinate,
malonate and propionate, details can be obtained from lab manual
(Wright, 2005).

The reaction times were recorded. To calculate the rate of reaction in
moles DPIP used per minute the formula in lab manual was used (Wright,
2005).

GRAPH

Discussion

Effect of pH on enzyme activity

The pH that makes the enzyme most active is known as the optimum pH.
The optimum pH for most enzymes is between 6-8. A change in the
optimal pH will interfere with some of the chemical bonds, altering
the shape of the enzyme and the active site as well to the point where
the substrate molecule could no longer fit, and the chemical change
would be inhibited from taking place as efficiently or not at all. If
the pH is very high or very low, enzymes can be denatured.

As shown in table 1. succinate was most active at a pH of 5 (3.5
mins) indicating that this pH is close to its optimal pH, whereas a pH
of 9 ( no change) denatured the enzyme rendering it inactive. A pH of
7 increased the activation time (4.3 mins) by 58 seconds from the time
recorded for a pH of 5. This increase in activation time indicates
that the enzyme prefers a lower pH than that of most enzymes (6-8)..

Effect of temperature on enzyme activity

The activity of enzymes increases when the temperature increases (and...

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