8. Answer the following questions. (6+3+3 pts) a Acid phosphatase catalyzes hydr

Question

8. Answer the following questions. (6+3+3 pts) a Acid phosphatase catalyzes hydrolysis of 4.0 umol of pNPP in 5 minutes when 2.5 ug of acid phosphatase is present in 5 mL. Calculate the i rate of reaction, ii enzyme activity and iii) specific activity b) Estimate k, the first order rate constant, for lysozyme when V 6.5 umole/L min and Km is 2.5 mM c) Determine the KM using the data shown in the plot below: 0.5 [S] mM a) 0.1 mM b) 0.2 mM c) 2.0 mM d) 4.0 mM e) Km cannot be determined from this data

Explanation / Answer

8. For the enzymatic reaction,

a)Calculating

i) rate of reaction of substrate concentration (M)/time = 4 micromol/0.005 L x 5 min = 160 microMolar/min

ii) Enzyme activity = 4 micromol/5 min = 0.8 micromol/min

iii) specific activity = 4 micromol/0.0025 mg x 5 min = 320 micromol/mg.min

b) rate constant = (0.0065 millimol/L.min)/(2.5 millimol/L) = 2.6 x 10^-3 min-1

c) Km using the plot

Km is substrate concentration at Vmax

Answer : b) 0.2 mM

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