On Fri, 28 Jan 2000 14:32:02, Kresten <kresten at my-deja.com> wrote:
ð I am looking for a thermostable protease preferably working around pH 5.
ð Does anybody know any proteases fulfilling these requirements? I do not
ð need specificity, actually the less specific the better.
ð
ð With regards to the temperature range, the bigger the better is my
ð answer at the moment although I might be satisfied by a protease working
ð around 70 degreesC. Also it would be nice if the protease was
ð commercially available.
You're in luck! Proteinase K, available from many sources, performs optimally
at 65 deg. C. A typical protocol for *complete* proteolysis would be 0.1 mg
proteinase/mL, 0.1-1.0% SDS. (The enzyme also tolerates urea up to 3-4 M, and
EDTA to 0.1 M or more.) There isn't much self-digestion at room temperature,
but you should keep the enzyme frozen when not in use. It's a serine esterase
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