Bio Lab Essay K101
The reactions and Results of Using Enzyme Turnip Root Peroxidase
Lab results from: Andrew Compton, Mickey. Results published 9/29/2012. TA:
In this series of laboratory experiments, my lab partner and I were to conduct an experiment about the oxidation rate of the enzyme peroxidase in the presence of its substrate guiacol. Also we used other substrates, such as hydroxylamine an enzyme inhibitor, to observe the weather the reaction rate was slowed down, sped up, or stopped reactions all together. These results were recorded by taking the materials in a test tube, then inserting them into a spectrophotometer to record the oxidation (intensity of color change from clear to reddish-brown) over a course of two minutes to observe color change. After all of the experiments that we ran we could conclude the enzyme oxidation rate (mmoles/min) depending on the amount of each product that was used in a single cubit test tube.
The following materials and Concepts were used to achieve oxidized/non-oxidized guaiacol:
* Boiled peroxidase (show the results of denatured enzymes/proteins when guaiacol is added) * Guaiacol (toxic substrate, common byproduct of cellular respiration) * Hydroxylamine (an extremely toxic carcinogen that is an enzyme inhibitor to peroxidase) * Peroxidase (enzyme from Turnip Root) * PH buffer of PH₅ * Spectrophotometer (record results of oxidation rate over two minutes.
The main objective of this lab was to observe the activity of enzyme peroxidase in real time under different experimental conditions. To see how peroxidase reacts with its substrate guaiacol under different conditions. We measure the amount of substances per test tube and then combine all of them together. Each test tube contains a measured amount of any listed substances including H₂O. Each measured amount of peroxidase along with its substrate guaiacol, and other listed substances will show how actively the enzyme oxidizes the substrate. To measure the
Lab results from: Andrew Compton, Mickey. Results published 9/29/2012. TA:
In this series of laboratory experiments, my lab partner and I were to conduct an experiment about the oxidation rate of the enzyme peroxidase in the presence of its substrate guiacol. Also we used other substrates, such as hydroxylamine an enzyme inhibitor, to observe the weather the reaction rate was slowed down, sped up, or stopped reactions all together. These results were recorded by taking the materials in a test tube, then inserting them into a spectrophotometer to record the oxidation (intensity of color change from clear to reddish-brown) over a course of two minutes to observe color change. After all of the experiments that we ran we could conclude the enzyme oxidation rate (mmoles/min) depending on the amount of each product that was used in a single cubit test tube.
The following materials and Concepts were used to achieve oxidized/non-oxidized guaiacol:
* Boiled peroxidase (show the results of denatured enzymes/proteins when guaiacol is added) * Guaiacol (toxic substrate, common byproduct of cellular respiration) * Hydroxylamine (an extremely toxic carcinogen that is an enzyme inhibitor to peroxidase) * Peroxidase (enzyme from Turnip Root) * PH buffer of PH₅ * Spectrophotometer (record results of oxidation rate over two minutes.
The main objective of this lab was to observe the activity of enzyme peroxidase in real time under different experimental conditions. To see how peroxidase reacts with its substrate guaiacol under different conditions. We measure the amount of substances per test tube and then combine all of them together. Each test tube contains a measured amount of any listed substances including H₂O. Each measured amount of peroxidase along with its substrate guaiacol, and other listed substances will show how actively the enzyme oxidizes the substrate. To measure the
References: Campbell, N and Reese, J.B (2006) Biology, p.142-149, Pearson/Benjamin Cummings, San Francisco CA Marrs, K (2007) K101 Laboratory Manual, Ex.5 “Characterization of Turnip Root Peroxidase” KhanAcademy (2012) “Oxidation and reduction cellular respiration” http://www.khanacademy.org/science/biology/cellular-respiration/v/oxidation-and-reduction-in-cellular-respiration.