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The effect of Hydrogen Peroxide substrate concentration on t

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Shivani Tumukuntala

on 16 September 2015

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Transcript of The effect of Hydrogen Peroxide substrate concentration on t

Source One: "the concentration of enzyme is maintained ...reaction rate will increase ...substrate is increased...some point...increasing substrate does not increase the reaction rate."
Source Two: "enzyme activity by out-competing competitive inhibitors that block enzyme ...protecting the enzyme against harsh pH environments."
Source Three: " As substrate concentration increases, the enzyme becomes saturated with substrate. As soon as the catalytic site is empty, more substrate is available ...rate of formation of product now depends on the activity of the enzyme...adding more substrate will not affect the rate of the reaction."

Revisit Purpose
Tested how various concentrations of substrate and affects upon the rate of enzyme activity
It was found that the more concentrated the substrate, the more enzyme activity there was, up to a certain point.
The effect of hydrogen peroxide substrate concentration on the rate of enzymatic activity of peroxidase.
Novel Application
Enzymatic reactions are constantly occurring in our bodies, hence understanding how factors can influence the rate of reaction is crucial. Helps us understand disorders that may be caused by failure in enzymatic activity by various outside factors. Doctors may be able to use this information to diagnose diseases and treat them.
To determine the effects of different substrate concentrations upon the rate of enzymatic activity.
Null Hupothesis:
The increase in substrate concentration has no effect on the rate of enzymatic activity.
Alternative Hypothesis:
If there is an increase in substrate concentration, then there is an increase in rate of enzyme activity up until a certain point.
Revisit Hypothesis
Future Question
15 mL of peroxidase was obtained
paper disc soaked in peroxidase and stuck onto the inside of the reaction chamber
Create Hydrogen Peroxide solutions with 10 mL of solution
Solution A: 0%
Solution B:1.5%
Solution C: 3% (control)
Solution D:4.5%
Solution E: 6%
10 mL of a hydrogen peroxide solution was placed into the reaction chamber
a pan was filled completely with water
50 mL graduated cylinder was submerged upright
reaction chamber was placed into pan of water(with the disks facing upward)
mouth of the graduated cylinder was placed over the top of the chamber tubing
reaction chamber was rotated
gas levels were measured in 30 second increments for 10 minutes.
Repeat with rest of solutions
6% Hydrogen Peroxide Solution
Distilled water
50 mL peroxidase
Reaction chamber
6 Droppers
Fall in line with previous research
Source One clearly shows that, provided enzyme concentration is kept constant, increased substrate will increase enzymatic activity, up until a maximum level of enzymatic activity is reached and it levels off.
The graph in Source One shows the same trends as our data.
Source Two also matches our findings- the higher concentration of hydrogen peroxide we used, the more enzymatic activity was observed via oxygen evolved.
Source Three supported as it showed that, as substrate increased, so did enzymatic activity initially, before leveling off, as seen in our first line graph.
As time increased, mL of O2 evolved also increased
a positive correlation between the two.
The rate of O2 produced
highest: beginning of the experiment (0-2 second range)
leveled off: 8 seconds
the higher the concentration of hydrogen peroxide in a solution, the steeper the rate of O2 produced
The 0% solution was the exception
3% hydrogen peroxide
the original solution
no increase or decrease in substrate concentration
controlled variables
total mL of substrate solution
the amount of peroxidase
time intervals of 30 seconds

0%: 6:00 there is a 0.1 mL of oxygen gas but it was predicted to have no mL of oxygen all throughout.
From an air bubble in the graduated cylinder that wasn’t taken out before inverting.
Prevented by moving the water in graduated cylinder back and forth to take out all the bubbles present.
3.0%: at time of 6-8 minutes a spike in mL of oxygen gas.
From a gas bubbles which were trapped in the pipe as the graduated cylinder was pressing on pipe and in between the time of 6-8 this was dislodged
Can be prevented by now placing pressure on the graduated cylinder when holding
Due to the fairly slow speed of our reactions it is only possible to measure the time of the reaction to the nearest 0.1 second even though the stopwatch shows the measurements to the nearest 0.01 second.
For overall effectiveness the experiment could be repeated more times

How does an increase in substrate concentration affect enzymatic activity in plant vs animal cells?
What is the highest concentration point to be reached by Hydrogen Peroxide for the rate of reaction to slow down?
What is the maximum possible rate of reaction?
How does change in substrate concentration affect rate in enzymatic activity at different pressures?

supported alternate hypothesis

increased substrate concentration from 0% to 6% caused the rate of enzymatic activity to increase with every interval.

rejected null hypothesis

our data clearly shows that the trend with increased substrate concentration is an increased rate of enzymatic activity.

Higher concentration of hydrogen peroxide resulted in an increase in oxygen production
More substrate molecules were present to collide with the active sites of the enzymes and thus they reacted at a faster, more efficient rate.
The highest peak of enzymatic activity during the first 0-2 minutes of each trial.
As the number of available active sites depleted, the rate of oxygen production gradually declined.
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