enzyme activity

[Arunas]

Hey everyone,

 

I'm doing a research on bacteria (color is reddish), that can digest coumarin. I sprayed 3-methylcatechol solution on those bacteria and after several hours bacteria became more yellow than reddish. It means that bacteria have 3-methylcatechol 2,3-dioxygenase (extradiol) activity. BUT I can't find any activity in their cellular extract. Due to this problem I can't purify this protein (molar weight also is unknown) and, furthermore, I can't get genes of coumarin digestion (we think that this dioxygenase participates in coumarin biotransformations).

 

So, any idea why I can't find enzyme activity in cellular extract?