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, 8.50 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 300 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 0.730 cm. The fraction with the highest activity had A540 of 0.743. The standard of 14.00 mM hydrolyzed sucrose had A540 of 1.087 (the concentration given is of the 100 µL sample). The no-enzyme control had A540 of 0.139. Calculate the activity of the fraction (in nmol min-1).
Enzyme kinetics
In biochemistry, enzymes are proteins that act as biological catalysts. Catalysis is the addition of a catalyst to a chemical reaction to speed up the pace of the reaction. Catalysis can be categorized as either homogeneous or heterogeneous, depending on whether the catalysts are distributed in the same phase as that of the reactants. Enzymes are an essential part of the cell because, without them, many organic processes would slow down and thus will affect the processes that are important for cell survival and sustenance.
Regulation of Enzymes
A substance that acts as a catalyst to regulate the reaction rate in the living organism's metabolic pathways without itself getting altered is an enzyme. Most of the biological reactions and metabolic pathways in the living systems are carried out by enzymes. They are specific for their works and work in particular conditions. It maintains the best possible rate of reaction in the most stable state. The enzymes have distinct properties as they can proceed with the reaction in any direction, their particular binding sites, pH specificity, temperature specificity required in very few amounts.
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, 8.50 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 300 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 0.730 cm. The fraction with the highest activity had A540 of 0.743. The standard of 14.00 mM hydrolyzed sucrose had A540 of 1.087 (the concentration given is of the 100 µL sample). The no-enzyme control had A540 of 0.139. Calculate the activity of the fraction (in nmol min-1).
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