optical density was measured at 270 nm for enzyme assay after sample collection time interval of 15 sec
Let's see if I got your question right. You must have some standard concentration of enzyme to calculate the unknown concentration. So far as I know, for enzymes, one must use the theory that if this much enzyme (known concentration) is converting the substrate into product (indicated by optical density of solution) in this time period that the solution with unknown enzyme concentration must have this much conc. of enzyme.
Here is a handy tool that lets you use the protein sequence to estimate the OD of an enzyme.
https://web.expasy.org/protparam/
My understanding is that it uses the amount or aromatic residues in the chain to estimate an extinction coefficient, allowing one to get protein concentration in an experiment using beer's law, and without having to make your own standard curve.
Suleman, means I must hypothesize the amount by taking consideration of the previous data???/
No farhan, no previous data required. You must have a known concentration of an enzyme to make a standard curve. Then from that standard curve, you can easily estimate the unknown concentration of enzyme in solution.
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