P5380

Protease from Bacillus licheniformis

Type VIII, lyophilized powder, 7-15 units/mg solid

• Synonym(s): Proteinase from Bacillus licheniformis, Subtilisin A
• CAS Number: 9014-01-1
• Enzyme Commission number: 3.4.21.62 (BRENDA, IUBMB)
• EC Number: 232-752-2
• MDL number: MFCD00132092
• UNSPSC Code: 12352204
• NACRES: NA.54

Properties

• biological source: Bacillus licheniformis
• Quality Level: 200
• type: Type VIII
• form: lyophilized powder
• specific activity: 7-15 units/mg solid
• mol wt: 27 kDa
• purified by: crystallization
• solubility: 10 mM NaAc (pH 7.5) and 5 mM CaAc: soluble, clear; H₂O: soluble, colorless
• foreign activity: DNase ≤5.0 Kunitz units/mg solid; RNase ≤0.05 Kunitz units/mg solid
• storage temp.: −20°C

Description

General description

The enzyme is found to be stable at pH 8-10, retaining activity of up to 90% for 24 hours. It shows maximum activity at temperatures between 55-60 °C.

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.

Application

Non-specific Protease Activity Assay Video

The product has been used with other enzymes for in situ proteolysis to produce crystals suitable for structure determination. It has also been used in the process of isolation of subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondria that can be used for functional in vitro studies.

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.

Biochem/physiol Actions

Protease catabolizes proteins by hydrolysis of peptide bonds. Proteases are inactivated by serine active-site inhibitors, such as phenylmethylsulfonyl fluoride (PMSF) and diisopropylfluorophosphate. Protease, Product P5380, is a serine endoproteinase with a broad specificity towards native and denatured proteins, and is active under alkaline conditions. It is active in some organic solvents such as dry octane.

Physical properties

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

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).

Safety Information

• Pictograms: GHS08,GHS05,GHS07,GHS09
• Signal Word: Danger
• Hazard Statements: H302,H315,H318,H334,H335,H410
• Precautionary Statements: P261 - P273 - P280 - P301 + P312 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Acute Tox. 4 Oral - Aquatic Acute 1 - Aquatic Chronic 2 - Eye Dam. 1 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Target Organs: Respiratory system
• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves