You are performing an enzyme reaction that works the same way as your wheat germ acid phosphatase (WGAP) assay. First, you dissolve 10.0 umol of product into 1.00 ml of buffer, and measure an absorbance of 0.500. Your cuvette has a 1.00 cm pathlength. Next, you perform the enzyme assay. Your enzyme solution has a concentration of 20.0 ug/ml. 1) Mix 0.250 ml substrate buffer with 0.250 ml enzyme 2) Incubate 20.0 min 3) Stop the assay with 0.500 ml NaOH 4) Read an absorbance of 0.200 *The units required for each answer in this problem are given. Answers to all three parts should be rounded to a maximum of three decimal places. Do not enter any letters. What is the extinction coefficient (ml/umol/cm) of the product? What is the catalytic activity (umol/min) measured in your assay? What is the specific activity (umol/min/ug) measured in your assay?
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.
You are performing an enzyme reaction that works the same way as your wheat germ acid phosphatase (WGAP) assay.
First, you dissolve 10.0 umol of product into 1.00 ml of buffer, and measure an absorbance of 0.500. Your cuvette has a 1.00 cm pathlength.
Next, you perform the enzyme assay. Your enzyme solution has a concentration of 20.0 ug/ml.
1) Mix 0.250 ml substrate buffer with 0.250 ml enzyme
2) Incubate 20.0 min
3) Stop the assay with 0.500 ml NaOH
4) Read an absorbance of 0.200
*The units required for each answer in this problem are given. Answers to all three parts should be rounded to a maximum of three decimal places. Do not enter any letters.
What is the extinction coefficient (ml/umol/cm) of the product?
What is the catalytic activity (umol/min) measured in your assay?
What is the specific activity (umol/min/ug) measured in your assay?
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