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Enzyme Kinetics Lab (Variables (substrate concentration (exhibits…
Enzyme Kinetics Lab
Variables
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time
more time passes, reaction increases until the substrate becomes a limiting factor/reaction reaches equilibrium
approach to equilibrium, but not its position, is accelerated by enzymes
pH
determine the optimal pH of the enzyme (best at 5 pH) makes sense because at alkaline pH it gets denatured
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Background about Enzymes
Enzyme Assay
mixing an enzyme of interest
with a substrate at a particular set of conditions, and then measuring the amount of
reaction that has occurred over a particular amount of time, to determine the effect of
enzymes on reaction rates.
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importance of stopping the reaction RIGHT before measuring in order to ensure that the timing is correct (to determine velocity/proper rate)
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Effects of Inhibitors
Competitive
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competes for ability to bind to the free enzyme (can't bind to the enzyme at the same time as the substrate)
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Background for the Lab
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product (pNP) turns yellow in basic conditions and the reaction stops in this pH range (enzyme is denatured
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