specific activity(IU/mg protein)?

Question

ive done the enzyme essay expt both purified and crude ones.
i need to get specific activity and i have done it
bt i am not so sure about it so plz check my answers
and if its wrong plz correct me!!

Activity of the purified enzyme
ΔAbs /min of 0.538
[got if from the spec with 400nm]
From Beer-Lambert Law A= εcl;
0.538=18300c ∴ c= 0.0000294 mol/l =0.0294mmol/l =0.0294umol/ml
The cuvette volume was 4mls
∴ 0.1ml AP transformed 0.1176umol pNPP per min
∴ 1ml AP 1.176 umol pNPP per min.
∴ the activity of the enzyme is 1.176 IU/ml enzyme.
Activity of the crude enzyme
ΔAbs /min of 0.320
A= εcl → 0.320=18300c ∴c=0.0000175 mol/l = 0.0175mmol/l=0.0175umol/ml
The cuvette volume was 4mls
∴ 0.1ml of crude enzyme transformed 0.07umol pNNP per min
∴ 1ml of crude enzyme 0.7umol pNPP per min
∴ the activity of the enzyme is 0.7 IU/ml enzyme



protein conc for crude is 13.6mg/ml
protein conc for purified is 0.05mg/ml

i dnt know if i need to use those protein conc figures!!

Answer 1

Suggest that you work backwards from what you want i.e.
enzyme specific activity. You will certainly need to take into account your protein concentrations in your calculations.
The crude enzyme protein concentration is much higher so that there's enough enzyme there to assay. The purified enzyme protein concentration is much lower otherwise you'd have too much enzyme. Suggest include the 'dilution factor '!

Answer 2

i'm grateful for having my comprehensive well being and being completely abled and having all 5 senses. i'm grateful that i become born and stay in a typical worldwide united states of america and dont ought to stroll miles for water, i only turn on the faucet. Im grateful for being captivating, its delivered me issues much less captivating human beings ought to never get so certainly :p

Answer 3

whats this?