o a points) The colorless the effect of experiment investigates the activity of
ID: 223472 • Letter: O
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o a points) The colorless the effect of experiment investigates the activity of an enzyme of hydrogen with the to a colored form production of tration will be presence oxidation, water and catalyzes breakdown changed from of oxygen. Certain the from a a hydrogen peroxide and that allows dyes will go coloriess dye a yellow oxidase, orho of amine (OPD) detection compound as it is oxidized H.0. (colorless) (reduced form) Using a spectrophotometer, from the OPD you quantitated the enzyme reaction by measuring the absorbency faster at The more colored OPD produced in a given time period, the and the enzymatic reaction. The following are the contents of Table 1) the absorbance readings al test tubes Absorbency HYDROGEN ENZYME at 410mm PEROXIDE (Potato (blank) 0.300 3.1 0.1 0.2 s 0.1 0.3 1.250 3.0 0.1 A. Write a hypothesis for this experiment (1 point) B. What is the substrate of the enzyme peroxidase in this experiment? (I point) c. which test tube contains the experiment designed as "no enzyme control" (see Table 1? point)Explanation / Answer
Answer A:
Peroxidase enzyme catalyzes the transfer of electrons from hydrogen peroxide to a colorimetric indicator. Its function is to break down hydrogen peroxide into water and oxygen, and in the presence of oxygen, o-phenylenediamine is first oxidized to the reactive o-quinone diimine, which reacts with another molecule of o-phenylenediamine to form 2,3-diaminophenazine (yellow-brown color). 2,3-Diaminophenazine was detected as a product of peroxidase-catalyzed oxidation of o-phenylenediamine.
Answer B:
As the enzyme name indicated, peroxidase enzyme use peroxides as substrate. Therefore, in the present experiment, hydrogen peroxide acts as a substrate for enzyme peroxidase.
Answer C:
Tube 1 (Blank) is considered as “no enzyme control” because it contains all the reagents except enzyme.
Answer D:
Buffer (PBS) is the independent variable because the amount of buffer can be varied to keep the total volume constant (3.5 ml).
Enzyme serves the dependent variable because the rate of oxidation of o-phenylenediamine is only dependent on the amount of enzyme present into the tube.
Answer F:
Competitive inhibition is a form of enzyme inhibition where binding of the inhibitor to the active site on the enzyme prevents binding of the substrate and vice versa. At any given moment, the enzyme either may be bound to the inhibitor or the substrate, but it cannot bind both at the same time. In competitive inhibition, a molecule similar to the substrate but unable to be acted on by the enzyme competes with the substrate for the active site. Because of the presence of the inhibitor, fewer active sites are available to act on the substrate. But since the enzyme's overall structure is unaffected by the inhibitor, it is still able to catalyze the reaction on substrate molecules that do bind to an active site.
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