After several purification methods, 57.75 µg of the enzyme alkaline phosphatase was purified from a crude extract and was found to hydrolyse 14.5 μmols of the substrate p-nitrophenyl phosphate to its products in 8 mins. What is the specific activity of alkaline phosphatase in units μmol min-1 mg-1.
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Since 14.5 μmols of substrate were hydrolyzed in 8 minutes, we can calculate the rate per minute: \[ \text{Rate} = \frac{14.5 \, \text{μmols}}{8 \, \text{min}} = 1.8125 \, \text{μmols min}^{-1} \] Show more…
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You are performing an enzyme reaction that works the same way as your wheat germ acid phosphatase (WGAP) assay: First, you dissolve 50.0 umol of product into 1.00 ml of buffer and measure an absorbance of 0.400. Your cuvette has a 1.00 cm pathlength. Next, you perform the enzyme assay: Your enzyme solution has a concentration of 10.0 ug/ml: 1) Mix 0.250 ml substrate buffer with 0.250 ml enzyme 2) Incubate for 10.0 min 3) Stop the assay with 0.500 ml NaOH 4) Read an absorbance of 0.100 The units required for each answer in this problem are given: Answers to all three parts should be rounded to the appropriate number of significant digits. What is the extinction coefficient of the product? ml.umol/cm What is the catalytic activity measured in your assay? umol/min What is the specific activity measured in your assay? umol/min/ug
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