A)Calculate specific activity of each fraction. B) what fraction of likely to co
ID: 208952 • Letter: A
Question
A)Calculate specific activity of each fraction. B) what fraction of likely to contain purest enzyme 8. You have discovered a new enzyme you name leprechaunase. In the purification of leprechaunase, three fractions from a chromatographic step were collected. Y activity. You also take 0.5 mL of each to determine the protein concentration. Your assays yield the following results that catalyzes the conversion of lead into gold that ou take 200 uL of each and assay them for racon Vox (mmoles/sec) Protein Conc. (mg) 10 20 25 10 10 20 A. Calculate the specific activity of each fraction. B. Which fraction is most likely to contain the purest enzyme? 8. Which of the following substances can behave as either an acid or a base, depending on the circumstances? (a) CHsSH (b) Ca (c) NH3 (d) CH SCHs (e) NH4 (0) H2C-CH2 (g) CH3CO2 (h) "HsNCH2CO2Explanation / Answer
A) In the question unit of substrate converted (Vmax) is in mmoles/200µL/sec. We need to convert it in µmol/mL/min to calculate enzyme unit.
For Fraction 1 substrate converted = 10mmol/200µL/sec= (10 x 1000) µmol/200µL/sec = 10000 µmol/0.2mL/sec = 50000 µmol/mL/sec = 3000000 µmol/mL/min
Similarly for Fraction 2 substrate converted = 20mmol/200µL/sec = 6000000 µmol/mL/min
Similarly for Fraction 3 substrate converted = 25mmol/200µL/sec = 7500000 µmol/mL/min
Now, assay volume = 200µL = 0.2 mL
Enzyme units= Assay volume (in mL) x substrate converted (in µmol/mL/min)
Now, specific activity of enzyme = Number of units of activity per unit of total protein
= Enzyme units / [Volume (mL) x Protein concentration (mg/mL)]
B) As the fraction 2 has highest specific activity so it will most likely contain the purest enzyme as more specific activity means more purity.
Fraction Calculation Enzyme Units 1 0.2 x 3000000 600000 2 0.2 x 6000000 1200000 3 0.2 x 7500000 1500000