I need some cheap model enzymes for kinetic studies. They should differ in their pI from acidic to basic. Substrates should be easily available. Product analysis should be possible by UV/Vis spectroscopy.
Thank you for the answers!
Look through the supplier catalogs for dehydrogenases. They can all be followed by the 340-nm absorbance change of NAD(P) NAD(P)H. Examples are lactic dehydrogenase and glucose-6-phosphate dehydrogenase.
Diaphorase with XTT and NAD(P)H.
There are various proteases with colorimetric peptide substrates (e.g. trypsin, chymotrypsin, etc.).
Beta-galactosidase with o-nitrophenyl-beta-D-galactoside, plus other glycosidases with their corresponding nitrophenyl substrates.
Nucleoside phosphorylase with MESG.
Once you have a list of enzymes, you will have to look up their pIs.
The most used enzyme used for Kinetic study was "Invertase" extracted from commercial yeast Saccharomyces cerevisiae.
The Substrate used was sucrose.
The enzyme was more active at pH 5.6
The products of the reaction (glucose and fructose) can be measured by DNSA (3,5 dinitrosalicilyc acid).
Rok Mravljak , There are a number of common enzymes you can utilize in this regard. In my opinion, the easiest one you can use is alpha-amylase, from the yeast, Saccharomyces cerevisiae. Its substrate is starch and you can decide to study its kinetics in terms of rate of maltose formed per unit time, at an absorbance of 540nm. Starch and maltose are readily available in most labs. Amylases have a fairly wide range of isoelectric points(3.25 - 10.1) depending on source.
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