I have carried out enzyme activity studies for strains A, B , C . Though the activity is different for all, the specific activity is almost same. So which data should I represent or will have more meaning? any explanation ?
Hi Rothangmawi, is your activity assay specific for a single enzyme or are there other enzymes that that can react to it?
If your assay is specific to a single enzyme, then specific activity (activity/protein conc.) is the best way to present the data.
There will be a problem if there are more than one enzyme that can be positive in your assay.
Hi there,
If you get different activities and the same specific activity, it means that your three extracts contain different protein concentrations. to evaluate the samples, you should consider specific activity which makes the link between the enzyme activity and it's relative abundancy in the lysate. But it might that your enzyme is subjected to different kind of regulation in your 3 strains. To get a better idea, you should also try to visualize the actual protein level in your samples (using WB for instance). Then you could make the real connection between activity and protein level.
Hope it helps.
Thank you Dominique for the reply. Yes indeed the protein concentration are very different, even thought I started with the same concentration of the sample. i guess i should try WB.
thanks
Hi Rothangmawi, is your activity assay specific for a single enzyme or are there other enzymes that that can react to it?
If your assay is specific to a single enzyme, then specific activity (activity/protein conc.) is the best way to present the data.
There will be a problem if there are more than one enzyme that can be positive in your assay.
Specific activity is very important when you go for protein purification. It depends on your research work that on what area you wish to concentrate? If you are just doing application work then enzyme activity is sufficient, but if you are going for purification specific activity becomes very important.
The specific activity (AS) is very important. It means that in all your samples, your protein is at the same concentration in all strains (let's say you have 1u/mg). If you knew the AS of the pure enzyme (let's say 100 U/mg) then you could calculate the real amount of your protein in the samples (here 10 µg of your protein in 1 mg total)
As Deepti has indicated, which is most important depends entirely on what you want to do. Do you want to USE this enzyme activity or to study the enzyme itself? If you want to use an activity e.g. in a crude extract it may be totally unimportant whether you have a tiny number of incredibly active enzyme molecules or a huge number of only moderately active molecules. On the other hand, if you want to study the enzyme for its own sake, or in your case, compare your A, B and C forms, then the specific activity (and lots of other things like substrate specificity, Km values, pH dependence etc. etc.) are really important.
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