I'm trying to assay for dioxygenase activity in Streptomyces (C12O, C23O,
PA34O, and PA45O) but I've been having some problems. First, I don't
have controls for ortho cleaving enzymes. Second, the spectrophotometric
method that I've been using for catechol 1,2-dioxygenase activity
(i.e. monitor for cis-cis-muconic acid formation at A260) hasn't been
working out due to the fact that I've been working with very crude
extracts. I found a colorimetric assay by K. Hosokawa which apparently
results in the formation of a purple color. Does anyone have any
experience with these assays? Also if anyone knows of or has any
Streptomyces or Pseudomonas (thought I'd prefer Streptomyces) isolates
that have ortho-cleavage activity I could really use them. If you have
any information please feel free to e-mail me:
Sakura Grace Iwagami
Dept of Microbiology
University of British Columbia
Vancouver, Canada
sakuragi at unixg.ubc.ca
or
sakura at davies.microbiology.ubc.ca
