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SRE0047

Sigma-Aldrich

Proteinase K from Tritirachium album

Suitable for manufacturing of diagnostic kits and reagents, lyophilized powder, ≥30 units/mg protein, for molecular biology

Synonym(s):

Endopeptidase K

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About This Item

CAS Number:
39450-01-6
Enzyme Commission number:
3.4.21.64 (BRENDA, IUBMB)
EC Number:
254-457-8
MDL number:
MFCD00132129
UNSPSC Code:
12352204
NACRES:
NA.54

biological source

fungus (Tritirachium album)

Quality Level

400

grade

for molecular biology

form

lyophilized powder

specific activity

≥30 units/mg protein

mol wt

28.93 kDa

impurities

≤1 ppm DNA

application(s)

diagnostic assay manufacturing

foreign activity

DNAse, Nickase and RNAse, none detected

shipped in

wet ice

storage temp.

−20°C

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General description

Proteinase K is an extracellular endopeptidase. It is synthesized by the mold Tritirachium album Limber. Proteinase K belongs to a new subfamily of the subtilisins. It is a 277 amino acid protein with a molecular weight of 28,930 Da. Active proteinase K is characterized with an unhydrolysed protein chain and autolysed polypeptide chains.

Application

Proteinase K catalyzes the hydrolysis of keratin.
Useful for the proteolytic inactivation of nucleases during the isolation of DNA and RNA.
Removes endotoxins that bind to cationic proteins such as lysozyme and ribonuclease A.
Reported useful for the isolation of hepatic, yeast, and mung bean mitochondria
Determination of enzyme localization on membranes
Treatment of paraffin embedded tissue sections to expose antigen binding sites for antibody labeling.
Digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research.

Biochem/physiol Actions

Proteinase K catalyzes the hydrolysis of keratin.
Proteinase K is a stable and highly reactive serine protease. Evidence from crystal and molecular structure studies indicates the enzyme belongs to the subtilisin family with an active-site catalytic triad (Asp39-His69-Ser224). It is stable in a broad range of environments: pH, buffer salts, detergents (SDS), and temperature. In the presence of 0.1-0.5% SDS, proteinase K retains activity and will digest a variety of proteins and nucleases in DNA preparations without compromising the integrity of the isolated DNA.

Unit Definition

One unit will hydrolyze urea-denatured hemoglobin to produce color equivalent to 1.0 μmole of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent).

Pictograms

Health hazardExclamation mark
GHS08,GHS07

Signal Word

Danger

Hazard Statements

H315,H319,H334,H335

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

Target Organs

Respiratory system

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

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Certificates of Analysis (COA)

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Marin Volarić et al.
Genes, 12(8) (2021-08-28)
The long-read Nanopore sequencing has been recently applied for assembly of complex genomes and analysis of linear genome organization. The most critical factor for successful long-read sequencing is extraction of high molecular weight (HMW) DNA of sufficient purity and quantity.
Proteinase K from Tritirachium album Limber
Ebeling W, et al.
FEBS Journal, 47(1), 91-97 (1974)
Amino acid sequence of proteinase K from the mold Tritirachium album Limber: Proteinase K ? a subtilisin-related enzyme with disulfide bonds.
Jany KD, et al.
Febs Letters, 199(2), 139-144 (1986)
Proteinase K from Tritirachium album Limber.
W Ebeling et al.
European journal of biochemistry, 47(1), 91-97 (1974-08-15)

Articles

Proteinase K: Introduction & Applications

Proteinase K aids in molecular biology applications by digesting structural proteins, removing nucleases, and isolating intact genomic DNA.

Proteinase K in Tissue Samples

Pro K aids in disrupting cell membranes for DNA release, crucial for downstream molecular biology techniques.

How Proteinase K Can Be Used in Blood DNA Extraction Applications

In blood DNA extraction, Proteinase K, an enzyme commonly used to degrade proteins, can help break down the cellular and nuclear membranes, releasing DNA from the cells that protect it from degradation and increase purity/yield making it more suitable for various molecular biology techniques.

Guide on How to Use Proteinase K in Different Procedures

Guidelines on use of proteinase K, an enzyme commonly used to degrade proteins, and protect DNA and RNA from degradation in samples.

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