Enzyme Lab

Purpose: To see what effect the concentration of the substrate has on enzyme activity (rate of reaction)
We used the enzyme catalase to quicken the breakdown of hydrogen peroxide, which can be dangerous
to the cell, into oxygen and water.
Background:
Enzymes are catalysts that aid in speeding up chemical reactions occurring in living cells. There is a
special region on the enzyme, called the active site that has a shape that fits with specific substrate
molecules. Because of this, each enzyme is only specific to a certain reaction. These substrate molecules
bind to the active site of the enzyme, where they may undergo a chemical change- forming a new
product. After releasing this product into the solution, the enzyme will return to its original shape, to
perform this action again and again. This process of substrate conversion would often times require
several chemical steps. Enzymes help to create something like a short-cut for these reactions. (see lab:
enzyme catalysis Adapted from 2006 AP workshop, Campbell AP biology book)


Procedure/ Set up:
We lined up all 18 beakers, (9 per trial) labeled them accordingly, and filled them with the appropriate
amount of room temperature water; 40 mL for the control, and then following the table for the rest of
them. We prepared the serial dilutions (also according to the table) with the hydrogen peroxide. Using
tweezers, 2 people would consecutively dip the filter paper cut-outs in the enzyme solution, dry it off,
and place it at the bottom of the beaker. Someone would then be timing while the last person recorded
the data. (see lab: enzyme catalysis Adapted from 2006 AP workshop)

Variables:
Independent: The enzyme catalase
Dependent: The substrate concentration
Control: Papers used, enzyme source, water source

Results: The data shows a clear trend in which the higher the concentration of hydrogen peroxide the
faster the filter paper would float to the top. The enzyme was more active at higher concentration
levels.




Data:
Substrate Concentration
(from serial dilution)
Time to Float (seconds)
Trial 1 Trial 2 Class Average Class Rate
0 (control) None None
0.1% hydrogen peroxide 46.3 52.13
0.2% hydrogen peroxide 27.66 23.93
0.3% hydrogen peroxide 20.9 17.6
0.5% hydrogen peroxide 13.06 13.3
0.8% hydrogen peroxide 9.01 9.08
1.0% hydrogen peroxide 6.6 2
2.0% hydrogen peroxide 4.41 3.6
3.0% hydrogen peroxide 2.65 4.6




Sources of error:
Timing using our cell phone stop watches is not going to be completely accurate- there was always at
least a +-.005 difference. It was slightly difficult to tell exactly when the disk reached the top, as well as
to properly get the disk on the bottom of the beaker to begin with. Because the enzyme solution (as well
as our groups hydrogen peroxide concentrations) took a while to prepare, we had some issues with
timing and had to rush towards the end. As always, we would have benefitted from doing more trials
and having more data to compare.

Discussion questions:
1) When increasing the concentration of the enzyme, the rate of reaction increases linearly as well;
concentration is doubled, rate of activity should essentially double too.
2) Increasing the concentration of the substrate will result in the enzymes rate of reaction increasing as
well. However, at some point all that excess substrate would not be able to find active sites on enzymes
available to bind to- the rate would then level out.
3) Once the substrate concentration was at 40.% hydrogen peroxide (and according to our data, well
before then) the rate would have stopped changing and become more constant
4) In our experiment,

5)

6) Enzymes are proteins, and can sometimes be sensitive to (especially high) temperatures. Each
enzyme has a optimum temperature at which is preforms its best. (http://www.hartnell.edu) For the
most part, as in all chemical reactions, the rate of activity will increase with heat- however, should the
temperature be too high, the enzyme can become denatured and inactive. The temperature groups
data showed that the paper took longest to float when the solution was 65+ degrees Celsius , so the
enzyme must have been reaching a temperature too high for it to work efficiently in.