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.[1] 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.[2]

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