I intend to use cyclooxygenase enzyme activity as an indicator of water contamination with NSAIDs but I require pre-concentration of the drugs. Pre concentration step with SPE includes solvent recovery of the drugs. Is Cox2 enzyme stable in methanol solutions? If so, what is the degree of stability?
I am wondering about the source of your enzyme, and the stability of the enzyme that you are handling considering that it is a hemoprotein.
Aromatic residues at the interior of globular proteins are exposed by methanol and other denaturing alcohols. Alcohols as denaturing agents act through hydrogen bonding or hydrophobic interactions. Globular proteins undergo conformational changes and assume a helical conformation on exposure to high concentrations of methanol.
http://www.jbc.org/content/245/10/2588.full.pdf+html
Thank you for your answers. I will use both Cyclooxygenase 2 from human and Cyclooxygenase 1 from sheep. I found some papers related to enzyme activity in organic solvents, but I could not find COX specific data.
I am considering immobilizing the enzyme, any thoughts on that?
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