Not available outside of the UK & Ireland.

Analysis Note

Amino acid analysis and isoelectric focusing electrophoresis consistent with subtilisin Carlsberg.

Application

This is a proteolytic enzyme isolated from the fermentation of Bacillus licheniformis. It is a serine endoproteinase with a broad specificity towards native and denatured proteins, and is active under alkaline conditions. Product P8038, also known as Subtilisin Carlsberg, has been used to hydrolyze cardiac cells to study the silencing of cardiac mitochondrial NHE1.

The enzyme from Sigma has been used to optimize release of all mitochondrial populations from homogenized ventricular tissue of rat heart. It has also been used in the pre-hybridisation treatment of formalin fixed, paraffin wax-embedded liver specimens for detecting human and viral DNA.

Biochem/physiol Actions

Proteinase catabolizes proteins by hydrolysis of peptide bonds. Proteases are inactivated by serine active-site inhibitors, such as phenylmethylsulfonyl fluoride (PMSF) and diisopropylfluorophosphate .

Packaging

1 g in glass bottle

50, 100, 250 mg in glass bottle

Physical properties

Subtilisin is a non-glycosylated single polypeptide chain without disulfide bonds and has a molecular weight of 27 KDa.

Specificity

Subtilisin A is a member of the Serine S8 Endoproteinase family. It has broad specificity with a preference for a large uncharged residue in the P1 position. It hydrolyzes native and denatured proteins, and is active under alkaline conditions.

Unit Definition

One unit will hydrolyze casein to produce color equivalent to 1.0 µmole (181 µg) of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent).