Would it be correct, if at some fixed concentration of substrate increasing enzyme concentration till the enzyme attain saturation, and then by dividing the corresponding enzyme activity of a saturation point by the enzyme amount at that saturation point.?
Would it be appropriate, prior knowing the Km, can we calculate the specific activity of enzyme.?
Thanks.
If you can figure out the rate (units), then divide it by mg of your enzyme SA=U/mg
Dear Irshad Baig:
The idea you suggested of increasing the enzyme concentration is close, but I think you would do better to increase the substrate concentration until you didn't get a significant increase in observed enzyme activity, then divide that activity rate by the concentration of enzyme. Specific activity for enzymes isn't exactly defined perfectly enough to say there is a "standard" way of doing it for all enzymes, but you'll get the most commonly defined way by doing it as suggested above.
Usually, the substrate is cheap and easy to get and the enzyme is hard, but if it is the other way around (for some enzymes it is easy and inexpensive to get the enzyme and difficult and expensive to get the substrate or run the assay) - if this is the case then one usually picks a standard concentration for the substrate (example would be 1 microgram/L, or micromole/L, or whatever concentration would be expected in vivo) and calculate it using this turnover rate.
Good luck, and I hope this addresses what your question really was.
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