Assay of thermostable proteins

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hmkvs
hmkvs's picture
Assay of thermostable proteins

Hello,

I have an enzyme from a hyperthermophile, I wanted to do the assay
of this protein. I have read that people incubate enzyme at different
temperatures and different times and do the assays for these at room
temperature itself. Suppose this protein is not at all active at room
temperature what should I do?
also the I need to use a coupled assay, so if I need to do the assay
at high temperature, the coupling enzymes wont support?
Can anyone give a possible soulution?

Thank you
hmkvs

Boopathi
Boopathi's picture
 You can use the temperature

 You can use the temperature which is slightly higher than the room temperature and below the optimum temperature for the enzyme. It should work for the both the enzymes without any problem.

g a
g a's picture
 Hi

 Hi

What are the optimum temperatures for the enzymes you are trying to use in  coupled assay

hmkvs
hmkvs's picture
Hello,

Hello,

Those enzymes has a temperature optimum of 37deg C. In
some of the papers they state that assays for the thermophilic
enzymes be better carried out at their actual temperatures,
because all thermophilic or hyperthermophilic enzymes will
not be functional at lower temperatures that we generally use.
Also it would be more relevent if the assays are carried out at
higher temperatures.

Thank you
hmkvs