A cellulase enzyme was purified and volumes in microliters were drawn to breakdown CMC as substrate in enzyme characterization studies. How do I calculate the enzyme concentrations in microliters or convert the enzyme volumes in microliters to concentration in micrograms?
You begin by measuring the protein concentration of the purified protein using any one of the usual protein assays. Then you just have to multiply the volume of the enzyme solution in µl by its concentration in mg/ml (= µg/µl) to get the number of µg used.
Measure the protein content in your enzyme extract either by Lowry method or Biuret method.
Calculate protein concentration in ug micrograms per ml of enzyme extract from BSA Bovine Serum Albumin standard curve prepared by using different concentrations of BSA. You can calculate the micrograms of protein in the volume of enzyme extract used for measuring activity.
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mg/mL protein conentration can be calculated to mol/L conentration using MW of the protein.
For example: If you multiply the mg/mL protein concentration with the reziprocal MW (Da) you will get the respective protein cencentration in M (mol/L).
A 10 mg/mL Lysozyme (MW of 14300 Da) solution would be 0,0007 M Lysozym.
Multiply it once again by 10^6 to convert M in µM. Hence a 10 mg/mL Lysozyme solution would be a 700µM Lysozym solution
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