Question

Can anyone help me with the following question? Thank you so much!

Biochemistry Lab 6: Protein Purification 0,07 0,06 0,05 0 po A280 Tot 0,04 0,03 0,02 0,01 Dం,000 tom, 0000000 0,00 10 20 30 Fraction nº LDH specific activity was only observed in fractions 21-26. Question #9: Based on the results of the A vs. fraction number and specific activity of those fractions, what can you conclude? What's going on with the big Ae peak at fractions 1-3? The peak at fractions 21-26? 40 i. ii.

Answer 1

1. LDH is the lactate dehydrogenase enzyme and helps in conversion of lactate into pyruvate and back. And we all know, enzymes are proteinases in nature and all enzymes work at a particular or specific temperature, pH etc. The question that why LDH specific activity is observed in fraction 21-26 because LDH show its maximum yield at this fraction.

Specific activity is always equal to total activity divided by total protein . It means that it's maximum yield or the amount of its activity retaination is maximum at this point after each purification step.

2. The above graph shows that when the LDH is being eluted at absorption of 280 NM with various fractions it show its specific activity at fraction 21-26 and the other peak at fraction 1-3 show that the enzyme slowactivity but less than the fraction 21 -26.

3. Further the small letters, a, b, c indicates that enzyme are not active at these particular fractions.