GRADE & PURITY EnzymoPure™, ≥500 units/mg dry weight

Accession #

P0C1U8

Bioactivity

≥500 units/mg dry weight

Overview

Protease S. aureus, V8 (endoproteinase-Glu-C) specifically cleaves peptide bonds on the COOH-terminal side of either aspartic or glutamic acids (Drapeau et al. 1972). Houmard and Drapeau (1972) report that in the presence of ammonium buffers the enzyme specificity can be limited to glutamoyl bonds. Its rather unique specificity, which can be considered as opposite to that of trypsin, makes it a useful tool for protein chemistry and peptide mapping studies (Cleveland et al. 1977) and (Hall et al. 1978).
Characteristics of Protease from S. aureus, V8:
Characteristics of Protease, Staph aureus (Endoproteinase Glu-C)
Molecular Weight
27,000 (Drapeau 1978).
Optimal pH
4.0 and 7.8 with hemoglobin substrate. (Drapeau et al. 1972).
Extinction Coefficient
extinction coefficient = 4.26 (Houmard 1976).
Inhibitors
Diisopropyl fluorophosphate (DFP) and monovalent anions such as F-, Cl-, Br-, CH3COO-, and NO3- (Houmard 1976).
Method
Enzyme activity is determined by the casein digestion assay described by Drapeau (1976). Since substrate grade caseins can vary from lot to lot and according to the manner produced, the standardization of casein digestion assays has been difficult to achieve. The use of a reference enzyme preparation is suggested. One unit is that amount of enzyme which releases acid soluble fragments equivalent to 0.001 A280 per minute at 37°C and pH 7.8 under the specified conditions.
Reagents
1% Casein in 0.05 M Tris⋅PO4 buffer, pH 7.8. (Dissolve 1 gram "Hammersten" casein in 50 ml 0.01 N NaOH with gentle heating and stirring. Add 40 ml reagent grade water and 5.0 ml 1.0 M Tris. Adjust pH to 7.8 with H3PO4 and q.s. to 100 ml.)
10% Trichloroacetic acid (TCA)
Enzyme
Dissolve at 1 mg/ml in reagent grade water.
Procedure
Equilibrate a series of tubes with 5.0 ml of 1% casein at 37°C for 5 minutes. At zero time add 10 ul or 20 ul of enzyme. Mix. Include a reagent blank. Exactly ten minutes after adding sample, stop reaction by adding 5.0 ml TCA. Mix. Allow tubes to stand ten minutes and then filter. Read A280 of the filtrate.
Calculation 

Specifications

Product Name

Endoproteinase Glu-C from Staphylococcus aureus V8, CAS No.66676-43-5

Synonyms

V8 Protease | Protease, Staph aureus from Staphylococcus aureus V8 | glutamyl endopeptidase | endoproteinase Glu-C | protease V8 | staphylococcal serine proteinase | Protease S Aureus | Glu-C Protease | GLU-C S.aureus

Grade

EnzymoPure™

Specifications & Purity

EnzymoPure™, ≥500 units/mg dry weight

Biochemical and Physiological Mechanisms

Preferentially cleaves peptide bonds on the carboxyl-terminal side of aspartate and glutamate. Along with other extracellular proteases it is involved in colonization and infection of human tissues. Required for proteolytic maturation of thiol protease Ss

Bioactivity

≥500 units/mg dry weight

Accession #

P0C1U8

Predicted molecular weight

27 kDa

CAS

66676-43-5

Enzyme Commission Number

3.4.21.19

Molecule Type

Enzyme

Storage and Shipping

Concentration

≥500 units/mg dry weight

Storage

Store at 2-8°C

Shipped In

Wet ice

Unit definition

One Unit causes a change of 0.001 A280 nm per minute at 37°C, pH 7.8 using casein as the substrate.

References

1. Wang Hongying, Li Zhiyan, Yang Wei, Chang Le, Yan Cunling.  (2025)  Development of commutable stabilizing control materials to improve external quality assessment of hemoglobin A1c assay.  ANALYTICAL AND BIOANALYTICAL CHEMISTRY,      [PMID:40736571] [10.1007/s00216-025-06018-4]