I am posting this request for a colleague who is unable to
post from his computer. Please send all responses directly to
him at the email address listed below.
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We are just beginning to look at oxidative stress in E. coli.
One of the assays we will use is that for catalase activity,
using the disappearance of H2O2 as an indicator. All the
protocols we have use the French Press to break the cells.
We will probably be precluded from this method due to the
manner in which we generate the external oxidant. (We
essential add a metal composite to the medium in which the
bugs are growing.) Would sonication be a viable alternative
or would this be somehow detrimental to the catalase? I have
used sonication before in other enzyme work (nitrogenase,
specifically) and it did not hurt the complex as far as we
could tell. If any one has any thoughts on this and/or can
point me in the right direction for recent reports on cell-free
catalase assays in bacteria, it would be greatly appreciated.
Please reply to rlsmith at albert.uta.edu.
THANKS.
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