Dynein Dynasty Team's Project: Difference between revisions
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Revision as of 20:21, 26 October 2013
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Background
As catalysts, enzymes serve to facilitate chemical reactions. Two specific enzymes, glucose oxidase and horseradish peroxidase, trigger related reactions in which one's products may act as the other's substrate. Specifically H202, the product of the reaction catalyzed by glucose oxidase, is also the substrate in a colormetric reaction which is catalyzed by horseradish peroxidase. Although the individual enzymes accelerate the reactions to a point, we believe that the velocity of the reaction and the rate of turnover can be further increased by binding the two enzymes together.
The Purpose (Mission)
Explore the possibilities of making enzymes more efficient by linking them together. The Project
In our project, we test the effect that crosslinking enzymes have on a colorimetric stepwise reaction. The horseradish peroxidase and glucose oxidase follow the stepwise reaction as below:
H20 +O2 + Glucose -GOx-> D-Glucose + ABTS + H202 -HRP-> ABTS (colored) + H20
It is hypothesized that the reaction will speed up since the products of the reaction with the first enzyme will already be near the site of its next reaction in which it is the reactants. Thus, the products of the first reaction do not simply wait until coincidentally meeting the next enzyme.
Methods
Azide-Alkyne Huisgen Cycloaddition
Azide-Alkyne Huisgen Cycloaddition is an example of "click chemistry" in which an azide from one molecule and aklyne from another react to form a triazole which connects the two previously separate molecules.
In our project, Azide Alkyne Huisgen Cycloaddition is used to connect Glucose Oxidase to Horseradish Peroxidase.
Crosslinking
Crosslinkning is the creation of a bond between two polymer chains (usually proteins) to connect the two. Crosslinking can be found in nature and also be used in biological and chemical research as it serves to strengthen a polymer. Once formed, crosslinks are difficult to break, explaining their usage in the vulcanization and durable objects such as tires.
