table shows show results obtained in a purification of lactate dehydrogenase fro
ID: 188580 • Letter: T
Question
table shows show results obtained in a purification of lactate dehydrogenase from crocodile muscle. Volume Protein Enzyme (mL) Conc. activity Specific Overall Yield (mg/mL) (U/mL) activity purification ( (U/mg) (% of starting activity) Stepl 50 10 400 n/a Crude extract Supernatant after 55 precipitation Step 2 340 Step 3 Pooled active fractions from anion exchange column 25 600 Step 4 Pooled fractions2 from coenzyme affinity column 6,000 With use of data shown in the table, calculate i) the specific activity (U/mg) at each stage, (ii) the overall purification achieved at each stage, (iii) the yield at each stage and as % of the starting activity. (2.5 marks) Write your answers in the table under the appropriate columns. a. 3 marks) (3 marks)Explanation / Answer
Ans a) Specific Activity : Toatal enzyme activiy divided by Total Protein
Overall Purification : IT is obtained by dividing the specific activity of any stage by the specific activity of orginal homogenate.
Yield : Can be calculated by multiplting the total volume of protein with enzyme activity.
Yeild % ; In this we consider the original homgenate as 100%. The yield can be calculated by dividing total activity of that step by the total activity of starting step( original homogenate).
Volume (ml) ProteinConc. (mg/ml) Enzyme activity(U/ml) Specific activity(U/mg) Overall purification yield(U) Yield( % of starting activity) Step1 50 10 400 400/10=40 n/a 50*400=20000 Step2 55 6 340 340/6=56.6 56.6/40=1.4 55*340=18700 340/400*100=85 Step3 25 1 600 600/1=600 600/40= 15 25*600=15000 600/400*100=150 Step4 2 2 6,000 6000/2=3000 3000/40=75 2*6000=12000 6000/400*100=1500Related Questions
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