Proteinase K From Engyodontium Album: P-Chloromercuribenzoate

Proteinase K
from Engyodontium album

(formerly Tritirachium album)
for Molecular Biology
Catalog Number P2308

Storage Temperature –20 C
2,3
CAS RN 39450-01-6 pH range: 7.5–12.0 (urea-denatured hemoglobin as
E.C. 3.4.21.64 substrate), but most often used in pH
Synonyms: Peptidase K, Endoproteinase K, range 7.5–9.0.
Endopeptidase K
Temperature profile: maximal activity at 37 C
3
Product Description (In the temperature range of 20–60 C, a minimum

Proteinase K is a stable serine protease with broad of 80% of the activity observed at 37 C is
substrate specificity. It degrades many proteins in the expected)
native state even in the presence of detergents.
Proteinase K was isolated from a fungus able to grow 2
pI: 8.9
on keratin and the enzyme can digest native keratin
1 2 1%
(hair); hence its name "Proteinase K". Evidence from Extinction coefficient: E = 14.2 (280 nm,
crystal and molecular structure studies indicates the 10 mM NaCl and 5 mM CaCl2, pH 8.0)
enzyme belongs to the subtilisin family with an
39 69 224 2+
active-site catalytic triad (Asp , His , and Ser ). The Activators: 1–5 mM Ca is required for activation.
predominant site of cleavage is the peptide bond When calcium is removed from the enzyme (by addition
adjacent to the carboxyl group of aliphatic and aromatic of EDTA), 25% of the catalytic activity is lost. However,
amino acids with blocked alpha amino groups. It is 2+
2-4
if the EDTA-Ca complex is removed from the enzyme
commonly used for its broad specificity. The mode solution by gel filtration, a total of 80% of the enzyme
4
and specificity of action has been studied. activity is lost and only a small activation will occur
2+ 2+ 16
upon addition of excess Ca to the Ca -free enzyme.
Proteinase K is frequently used in molecular biology
applications to digest unwanted proteins, such as Inhibitors: Proteinase K is inhibited by DIFP or PMSF
nucleases in DNA or RNA preparations from 3
5-11
(the latter used at final concentration 5 mM). It is
microorganisms, cultured cells, and plants. The partly inactivated, but not inhibited, by EDTA (see
enzyme is typically used at 50–200 g/ml in nucleic Activators). Proteinase K is not inhibited by iodoacetic
acid preparations at pH 7.5–8.0 and 37 C. Incubation acid, the trypsin-specific inhibitor TLCK, the
times vary from 30 minutes to 18 hours. Proteinase K chymotrypsin-specific inhibitor TPCK, or
is usually denatured by subsequent phenol extractions, p-chloromercuribenzoate.
3
although it can autodigest during long incubations.
Catalog Numbers P2308 and P4850 [a solution in 40% This product is supplied as a lyophilized powder.
(v/v) glycerol and Tris buffer] are tested for suitability in
molecular biology applications. Protein content: 90%
Proteinase K is active in 1% Triton X-100 and fully Specific activity: 30 units/mg of protein
active in 0.5% (w/v) SDS. SDS and urea will denature
protein substrates resulting in increased digestion rates. Unit Definition: One unit will hydrolyze urea-denatured
Proteinase K itself is denatured much more slowly by hemoglobin to produce color equivalent to 1.0 mole
3,12,13
these agents.
(181 g) of tyrosine per minute at pH 7.5 at 37 C.
14
Molecular mass: 28,930 Da (amino acid sequence)
15
28,500 Da (SDS-PAGE)
Impurities: References
DNase: none detected 1. Betzel, C. et al., Eur. J. Biochem., 178(1), 155-171
Endonuclease (nickase): none detected (1988).
RNase: none detected 2. Ebeling, W. et al., Eur. J. Biochem., 47(1), 91-97
DNA: 1 ppm (1974).
3. Enzymes of Molecular Biology, Vol. 16 (Burrell,
Precautions and Disclaimer M.M., ed.), Humana Press (Totowa, NJ), p. 307
This product is for R&D use only, not for drug, (1993).
household, or other uses. Please consult the Safety 4. Kraus, E., and Femfert, U., Hoppe Seylers Z.
Data Sheet for information regarding hazards and safe Physiol. Chem., 357(7), 937-947 (1976).
handling practices. 5. Lizardi, P.M., and Engelberg, A., Anal. Biochem.,
98(1), 116-122 (1979).
Preparation Instructions 6. Gross-Bellard, M. et al., Eur. J. Biochem., 36(1),
This product is soluble in water (1 mg/ml), yielding a 32-38 (1973).
nd
clear colorless solution. 7. Molecular Cloning: A Laboratory Handbook, 2 ed.
(Sambrook, J., et al., eds.), Cold Spring Harbor
Storage/Stability Press (Cold Spring Harbor, NY) p. 1.61 and p. B.16
Sigma recommends storage at –20 C, though others (1989).
have claimed stability at 2–8 C. When stored at 8. Kasche, V. et al., Prep. Biochem., 11(3), 233-250
–20 C, the product retains activity for at least 2 years. (1981).
9. Hansen, J.N., Prep. Biochem., 4(6), 473-488
A Proteinase K solution remains active over a broad pH (1974).
range (4.0–12.5, optimal pH 8.0) and also over the 10. Holm, C. et al., Gene, 42(2), 169-173 (1986).
temperature range of 25–65 C during use. At pH 8.0, 11. La Claire, J.W., and Herrin, D.L., Plant Mol. Biol.
solutions remain active for at least 12 months at Reporter, 15(3), 263-272 (1997).
12. Hilz, H. et al., Eur. J. Biochem., 56(1), 103-108
2–8 C. At pH 4–11.5, solutions containing Ca
3 2+
(1975).
(1–6 mM) are expected to remain active for several rd
13. Methods of Enzymatic Analysis, 3 Edition
weeks. An 80% ammonium sulfate suspension stored
(Bergmeyer, H.U., ed.), Academic Press (New
at 2–8 C remains active for at least 12 months.
2
York, NY), Vol. 2, p. 299 (1983).
14. Jany, K.-D. et al., FEBS Lett., 199(2), 139-144
(1986).
15. Jany, K.-D., and Mayer, B., Biol. Chem. Hoppe-
Seyler, 366(5), 485-492 (1985).
16. Bajorath, J. et al., Eur. J. Biochem., 176(2), 441-
447 (1988).
Triton is a trademark of The Dow Chemical Company or

an affiliated company of Dow.
SBC,GCY,RBR,RBG,MAM 10/18-1
2018 Sigma-Aldrich Co. LLC. All rights reserved. SIGMA-ALDRICH is a trademark of Sigma-Aldrich Co. LLC, registered in the US and other
countries. Sigma brand products are sold through Sigma-Aldrich, Inc. Purchaser must determine the suitability of the product(s) for their
particular use. Additional terms and conditions may apply. Please see product information on the Sigma-Aldrich website at
www.sigmaaldrich.com and/or on the reverse side of the invoice or packing slip.

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Proteinase K

from Engyodontium album

Catalog Number P2308

Product Description (In the temperature range of 20–60 C, a minimum

Triton is a trademark of The Dow Chemical Company or

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