Hallo,
Do you know a SAM-methyltransferase assay for a thermophilic methyltransferase?
I see a lot of kit, but they are for enzyme that works at 37°C
Can the enzyme reaction and the detection be done at different temperatures with these kits? In other words, what is preventing you from using the kit at the higher temperature?
Thanks for your reply, actually I do not know, but most likely my enzyme does not work at lower temperatures (or works less) while for the kits, the manufacturers tell me they have never been tested those temperatures
I looked at the G Biosciences kit. It is a continuous coupled system. It might be possible to separate the SAM methyltransferase reaction from the detection, making it a discontinuous assay. My idea is that you do the reaction of your enzyme at the high temperature it requires, then lower the temperature and add the detection reagents. The activity of your enzyme should be greatly reduced at the lower temperature, so the temperature reduction acts essentially as a quencher. Then, allow sufficient time for the coupling enzymes to complete the conversion of SAH and read the final absorbance.
Thank you for the answer!
It is what I was thinking of doing, 2 steps assay at 2 different temperatures.
thank you :)
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