A student performed an invertase activity assay on samples from a purification. All reactions occurred in a reaction volume of 100 µL and lasted 3.1 minutes. For each reaction, 5.00 µL enzyme was mixed with 50.0 mM sucrose in 10.0 mM acetate pH 4.8. Then 400 µL of DNS was added, the samples were heated to 90 °C for 5 minutes, and 500 µL of 50.0 mM acetate pH 4.8 was added before loading a 96 well plate in triplicate. Samples were measured at 540 nm using a path length of 0.680 cm. The fraction with the highest activity had A540 of 0.904. The standard of 14.00 mM hydrolyzed sucrose had A540 of 1.047 (the concentration given is of the 100 µL sample). The no-enzyme control had A540 of 0.174. Calculate the activity of the fraction (in nmol min-1).
A student performed an invertase activity assay on samples from a purification. All reactions occurred in a reaction volume of 100 µL and lasted 3.1 minutes. For each reaction, 5.00 µL enzyme was mixed with 50.0 mM sucrose in 10.0 mM acetate pH 4.8. Then 400 µL of DNS was added, the samples were heated to 90 °C for 5 minutes, and 500 µL of 50.0 mM acetate pH 4.8 was added before loading a 96 well plate in triplicate. Samples were measured at 540 nm using a path length of 0.680 cm. The fraction with the highest activity had A540 of 0.904. The standard of 14.00 mM hydrolyzed sucrose had A540 of 1.047 (the concentration given is of the 100 µL sample). The no-enzyme control had A540 of 0.174. Calculate the activity of the fraction (in nmol min-1).
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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A student performed an invertase activity assay on samples from a purification. All reactions occurred in a reaction volume of 100 µL and lasted 3.1 minutes. For each reaction, 5.00 µL enzyme was mixed with 50.0 mM sucrose in 10.0 mM acetate pH 4.8. Then 400 µL of DNS was added, the samples were heated to 90 °C for 5 minutes, and 500 µL of 50.0 mM acetate pH 4.8 was added before loading a 96 well plate in triplicate. Samples were measured at 540 nm using a path length of 0.680 cm. The fraction with the highest activity had A540 of 0.904. The standard of 14.00 mM hydrolyzed sucrose had A540 of 1.047 (the concentration given is of the 100 µL sample). The no-enzyme control had A540 of 0.174. Calculate the activity of the fraction (in nmol min-1).
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