DENNIS MUBAIWA

Write up of enzyme practical

Enzyme Concentration/%
10
8
6
4
2
0

Time taken for

reaction to reach end
point/s
240
232
223
304
276
312
338
331
334
402
398
403
550
557
545
>3000

Average/s
232
297
334
401
551

Enzyme
Concentration/%

Rate Of Reaction/ s- x
10-

Avg Rate Of
Reaction/s- x 10-

10

4.17

4.31

4.48

4.31

3.29

3.62

2.21

3.37

2.96

3.02

3.00

3.00

2.49

2.51

2.48

2.49

1.82

1.80

1.48

1.81

Our results were fairly reliable as they were not too far off each other in the
repeat experiments. 10% and 8% concentrations were all relatively around the
(220 < x < 320) zone. But as expected the reactions began to take longer as the
enzyme concentrations got less and less. This is evident as 4% concentration
was between 398 and 403 seconds but 2% was between 545 and 557 seconds.
The rate of reactions were inverses of the times multiplied by a thousand.
So what does all this show?
This trend therefore corresponds to and seems to support our hypothesis, which
was, as the enzyme concentration gets higher there are more substrates
available for collision therefore the rate of concentration is higher
The experiment showed that , by reducing the enzyme concentration, there were
less active sites available for the substrate to occupy. It was enzyme limiting.
This meant that there would be far less collisions occurring, therefore increasing
the amount of time it took for the reaction to complete.
Also, it was possible that the reactions would reach an end point whereby the
substrate has been completely expelled. This would explain the solution being

consistently cloudy, with no change being detected after a prolonged period of

stirring.
So what could have gone wrong?
Well since we are talking of reactions at a microscopic level, it is fair to
acknowledge that it is almost impossible to create a 100% reliable experiment.
However what is good to know is that these errors and uncontrolled variables
would not be significant enough to affect the validity of the whole experiment. Or
could they? Therefore I shall proceed to take note of the possible errors in the
procedure.
Also errors with timing could have been a large factor. If the concentration was
too low then the reaction reaches a point where all the active sites are occupied
but there is free substrate. This slowed down the rate of reaction, and nothing
seemed to happen. This was evident. There were cases where the solution
reaches a relatively high level of clearness but seemed to stay that way for
periods of up to 30 seconds. But this didnt particularly mean the reaction was
over, but then this was assumed to be the end point. Meaning the timing has
stopped while the reaction is still very much in process. This was more
systematic as it was the case with every experiment.
Since this was more of a judgemental experiment, lots of errors arose. How clear
did the solution have to be for the reaction to be regarded as complete? This
was a question that was answered individually within groups as they all used eye
judgement to determine the end point. But two eyes never see the same, so end
points were different. A colorimeter could have been employed to rectify this, but
it was not in this experiment. This meant it was impossible to stop the clock at
identical levels of clearness affecting the validity of the experiment. So this
caused fluctuations within time. I expected the reaction times to be directly
proportional to each other, which they seem to be, but only very slightly.
Also, stirring the solution was a factor. All solutions were stirred with varying
amounts of vigour, which would have either increased or decreased the collisions
within the solution.

Any Random errors?

The first errors could have arisen in the dilution of the enzyme, as we attempted
to bring down the concentration. This would be maybe mixing too much enzyme
with water, or maybe the reverse. This was evident as some groups complained
of shortage of enzymes, yet it was made sure that every group had sufficient
enzyme solution for 3 repeat experiments. This could swing the results either
way as it could for starters shorten the amount of time the reaction took place in,
or lengthen in relative to the error. This is random as it occurred in some groups,

but did not in others.

Also, another error, should I say variable that could have affected the results
was temperature. One thing is that body temperature is higher than room
temperature. The solution was at more or less room temperature and we handled
the containers repeatedly. This would have accelerated the reaction as the
particles reacting would have gained more energy through the heat and collided
more, however it would have been fine if we handled the solutions in exactly the
same way for the same amount of time. But we did not so this could have been
responsible for the fluctuation in the time it took to complete the reaction. This
was also random as it could have happened during one part of the experiment.
Also, contamination could have played a part in affecting the results validity. For
example adding 6% concentration acid in a container contaminated with a
stronger concentration now meant that the concentration was stronger than it
was meant to be. This would make the reaction go a lot quicker than it was really
supposed to. Also affecting the validity of the experiment.
So what about your results?
The results were reliable enough to show the trend that supported our
hypothesis. But were they reliable? No. Were they valid? No. So why was this?
As mentioned earlier there were too many errors and too many uncontrolled
variable that could have affected the whole experiment. By far the most crucial
and largest was judging the end point. The whole set up of the experiment
made it impossible to know the end point. Without a colorimeter or a way of
declaring a level of clearness to be dubbed as the end point, it was impossible to
judge accurately. This was evident with 8% concentration. One of the times was
276s and the other 312s. That was 36 seconds time difference on only one
concentration, and it was reflected by the huge error bars on the graph. The
graph showed a positive gradient, meaning that concentration affected the rate
of reaction. I believe a perfect experiment would have shown a completely
straight line, possibly a gradient of 1.
How can the results be improved?
This is difficult, as there are too many errors possible. To make the experiment
better, I propose pre-made enzyme concentrations. These would eliminate any
dilution problems. Also, the use of a completed reaction as a colorimeter would
come in handy. This can be used so that it can be compared with an ongoing
reaction to judge where the reaction ends. Also maybe carrying the experiment
in a temperature controlled environment. This would eliminate random errors
associated with sudden temperature changes. Some of the procedures are
difficult to replace, but these are the simplest to do without changing the
purpose and feasibility of the experiment totally.