Acetyl-CoA acetyltransferase- Activity study protocol - (Jan/03/2006 )

Hello Friends,

If anyone working on the Acetyl-CoA acetyltransferase gene from Clostridium or any other bacterial source , could you please share me the activity study procedure.I am looking for a standard protocl to measure the activity in wild and recombinant bacteria.Thanks in advance.

Sincerely

Saikat Chakraborty
Mie University
Japan

Can you mix your enzyme with some free cholesterol and 14C oleoyl-CoA let it incubate at 37C for 10-20 minutes, extract the cholesterol ester that is made, run it up a TLC plate, then scrape and count the cholesterol ester band.