At high pH, alpha and beta-cyclodextrin have been found to have cleave aromatic esters. What experiment would most definitively support this mechanism? How would one test the Kinetics of a enzyme-substrate reaction? At high pH, alpha and beta-cyclodextrin have been found to have cleave aromatic esters. What experiment would most definitively support this mechanism? How would one test the Kinetics of a enzyme-substrate reaction? @Chemistry
I'm assuming you're talking about an enzyme that cleaves alpha and beta-cyclodextrin, because alpha and beta-cyclodextrin are aromatic esters themselves. Since the enzymes function depending on the specific pH of the environment, a direct experiment that would support this mechanism would have a changing pH condition. For example add enzyme and substrate in varying pH conditions, then measure the rate of the disappearance of the substrate or the creation of the product as a function of the pH of the environment, if your hypothesis is true, the rate of reaction for of the enzyme will be optimal under (basic) high pH conditions. Plot reaction rate vs. pH to verify. Enzymatic kinetics can be measured by varying the concentration of the substrate with a fixed amount of enzymes and record the reaction rate as a function of the substrate concentration. This is known as the Michaelis–Menten kinetics. The curve will eventually plateau when all the enzymes are saturated with the substrate, where no faster rate of transformation can be achieved. To actually detect the cleavage of the aromatic esters you can measure the specific absorbance of the reaction system with the help of fluorescence spectroscopy, since aromatic compounds display unique emission characteristics, a decrease in the peak at this specific wavelength can indicate that the actual substrate is being consumed by the enzyme.
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