Reasons for enzyme purification? - (Jun/01/2006 )
This is a very general and fundamental question. I just recently purified lactate dehydrogenase from beef heart and am currently doing a report. However, when I wondered about what the AIM of purifying an enzyme was, I could only come up with a few reasons after some research and thought.
1.) To understand what is going on in a cell and between cells: reconstruction of metabolic and regulatory pathways. Pure enzymes/proteins required to study reactions, kinetics, regulation, etc.
2.) To understand deviations in normal metabolism or regulation processes, etc., due to abnormal enzymes/proteins (e.g. mutants of globins --> thalassemia's and sickle cell disease).
3.) To make a rational design of drugs possible, based on the 3D-structure of a protein.
4.) Many proteins/enzymes have themselves an added value, as biocatalysts (proteases, lipases, glucose isomerase), therapeutics (insulin, interleukins), etc.
5.) To study details concerning the purified enzyme e.g. 3D crystal structure, a.a. sequence.
I would appreciate it if you guys can help me add to the list!
ummmm maybe those answers are casting the net a little to wide
normally the aim, whould be what you set out to do at the onset of your experiment i.e. why isolate lactate dehydrogenase? what were you going to do with the enzyme once isolate? your aim is specific to your experiment
Maybe I wasn't very clear sorry. The AIM of the experiment I had was to purify lactate dehydrogenase from a crude tissue homogenate. But I am trying to explore into WHY do we purify lactate dehydrogenase or any other enzymes for that matter. The report i am writing is simply based on the purification part but not explaining why =S So perhaps I thought I could go into possible explanations of what implications there would be for obtaining a pure enzyme.
I would put that information in the discussion, and it seems pleanty of possible applcations as is