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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

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Synonym(s):
Endopeptidase K
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.54

biological source

fungus (Tritirachium album)

Quality Level

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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This Item
P8044P2308SAE0009
grade

for molecular biology

grade

-

grade

for molecular biology

grade

-

biological source

fungus (Tritirachium album)

biological source

maize, microbial (T.album
T. ALBUM), plant seeds (barley), soybean, yeast

biological source

-

biological source

fungus (Tritirachium album)

impurities

≤1 ppm DNA

impurities

-

impurities

≤1 ppm DNA

impurities

-

Quality Level

400

Quality Level

200

Quality Level

400

Quality Level

200

application(s)

diagnostic assay manufacturing

application(s)

diagnostic assay manufacturing

application(s)

-

application(s)

-

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

Exclamation markHealth hazard

signalword

Danger

Hazard Classifications

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

target_organs

Respiratory system

Storage Class

11 - Combustible Solids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


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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 is commonly used in molecular biology and biochemistry applications to digest structural proteins and enzymes. It is useful in removing nucleases that can degrade DNA and RNA, as well as in the isolation of intact genomic DNA from various sources.

The use of Pro K in combination with other reagents, such as detergents and chaotropic agents, can help to disrupt the cell membranes and release DNA from tissue. This is particularly important for downstream applications such as PCR, sequencing, and other molecular biology techniques that require pure and intact DNA.

The cost of proteinase K can vary depending on the source, purity, manufacturing process and vendor’s quality management system. It is important to balance the cost with the desired quality, performance, documentation and technical/quality support to select the optimal Proteinase K for the intended application

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.

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