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蛋白酶 K ( Proteinase K from Tritirachium album) C122917

2018-04-09

Catalog Number:C122917

Size:20mg/100mg 

CAS Number:39450-01-6 


Applications

 Proteinase K is suitable for both protein and nucleic acid isolation. Exhibits proteolytic activity on proteins, peptides, glycoproteins, amides and esters. Also active with nitroanilides of amino acids with protected amino groups, excluding arginine. Useful in the isolation of DNA and RNA, in the analysis of membrane structures and protein structure. Addition of either 0.5-1% of sodium dodecyl sulfate or 1-4 mmol/l of urea increases the activity, because the substrates are more easily attacked when denatured. In experiment to recover the undigested DNA, proteins were digested with 0.1 mg/ml of proteinase K. 

Product Description 

A highly active stable endopeptidase with a broad spectrum of action was isolated by E. Merk's Darmstadt Biochemical Research Department in 1970 from a culture filtrate of the fungus, Tritirachium album Limber, suitable for both protein and nucleic acid isolation, it exhibits proteolytic activity on proteins, peptides, glycoproteins, amides and esters. Also active with nitroanilides of amino acids with protected amino groups, excluding arginine. 

Biochem/physiol Actions

 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 Anson unit is the amount of enzyme which will liberate 1 mole of folin-positive amino acids per minute at pH 7.5 and 35°C using hemoglobin as substrate. 


Others: 

             Specific Activity:>30 mAnson u/mg 

             Synonyms:Endopeptidase K, Proteinase, Tritirachium album serine 

             Enzyme Commission Number:E.C.3.4.21.64 

             Molecular Weight:~30 kDa 

             EC Number:254-457-8 

             MDL Number:MFCD00132129 

             Keywords:Nucleic acid isolation, Endopeptidase K, 3.4.21.14. , Proteolytic Enzymes, Proteolytic Enzymes and Substrates, Peptidases, Hydrolases, Biochemicals and Reagents, Core Bioreagents, Enzyme Class Index, Enzymes, Enzymes, Inhibitors, and Substrates, Life Science Reagents for Cloning, Modifying Enzymes,Molecular Biology, Molecular Biology Enzymes, Plant Molecular Biology, Plant Nucleic Acid Purification

             Key Applications:Protein and nucleic acid isolation 

             Product Type:Proteins, Enzymes & Peptides 

             Biochemical Category:Proteases 

             Presentation:White to off white lyophilized powder Isoelectric point (pI):8.9 

             pH:pH optimum (denatured hemoglobin as substrate): pH 7.5-12.00 

             Specificity:Proteinase K cleaves peptide bonds mostly after the carboxyl group of N-substituted hydrophobic aliphatic and aromatic amino acids, as shown by specificity trials with amino acid-4-nitroacilides. Thus, it shows similarities with alkaline Asperigillus proteases. However, unlike the latter, Protease K also cleaves peptide amides, comparable to the alkaline serine-proteases from Bacillus species. The specificity of ester cleavage is also high. 

             Solubility:Soluble in water (1 mg/mL) 

             Storage & Handling:+4 °C, dessicate 

             Foreign activity: 

                                       RNase <5 × 10-4 u/mg D

                                       Nase <5 × 10-4 u/mg




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