Dear All
I am working on the retention (Adsorbtion) of different enzymes on the surface of different materials including soil, zeolite and etc. I need some information about the determination of enzyme concentration in solutions. I got some pure enzyme as powder form and should measure the capacity of this base material for retention of enzymes.
As long as the solid material does not contain any soluble protein and does not contribute a significant amount of colored material or turbidity to the liquid phase, then you can determine the amount of the added enzyme that remains in the liquid phase by using a protein assay. First, use a centrifuge to separate the solid material from the liquid phase. Then do the protein assay on the clarified liquid phase. Use the enzyme stock solution of known concentration as the standard. There are several protein assay reagents available commercially, including Bradford, BCA, Lowry, and 660 nm.
Hi there,
If the protein is pure then solution prepared from the pure powder will also be pure. The UV Spectrum of the solution will be specific of the presence of the protein at the defined concentration (provided the protein is sufficiently rich in aromatic aas). So comparing UV spectra of the liquid phases before and after treatment with your solid phase will tell you what proportion of protein is still in the liquid phase and therefore you may deduce how much is adsorbed on the solid phase.
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