thermostable proteases

Dr. Peter Gegenheimer PGegen at UKans.nolospamare.edu
Fri Jan 28 20:33:50 EST 2000


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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