A9934

Aminopeptidase I from Streptomyces griseus

lyophilized powder, ≥200 units/mg protein

• Synonym(s): Leucine Aminopeptidase IV
• CAS Number: 9031-94-1
• Enzyme Commission number: 3.4.11.22 (BRENDA, IUBMB)
• EC Number: 232-874-6
• MDL number: MFCD00166325
• UNSPSC Code: 12352204
• NACRES: NA.54

Properties

• form: lyophilized powder
• Quality Level: 200
• specific activity: ≥200 units/mg protein
• mol wt: 21 kDa by gel filtration; 33 kDa by SDS-PAGE
• composition: Protein, 40-60% Lowry
• storage temp.: −20°C

Description

General description

Aminopeptidase I from Streptomyces griseus is a thermostable enzyme with Glu131 and Tyr246 as key active site residues.

Application

Aminopeptidase I from Streptomyces griseus has been used:

• to test the biochar exposure effect on the enzyme activity
• in circular dichroism (CD) spectroscopy studies
• as a positive control in p-nitroanilide degradation assay

Biochem/physiol Actions

Aminopeptidase I from S. griseus has a fairly broad specificity, being able to remove the N-terminal residue of most proteins, except where the penultimate residue is an imino acid. It contains two Zn²⁺ binding sites. Aminopeptidase I from S. griseus is inhibited by 1,10-phenanthroline and is activated six-fold by Ca²⁺, which also stabilizes it against heat inactivation. This monomeric zinc metalloprotein has an isoelectric point (pI) of 5.4.

Aminopeptidase I may also be used as a reagent in the assay of endoprotease activities with a synthetic substrate in a two-stage assay. In the first stage, the endoprotease cleaves a peptide, such as Z-Y-X-Leu-p-nitroanilide, with the X, Y, and Z residues being chosen according to the specificity of the endoprotease.

Packaging

Package size based on protein content.

Unit Definition

One unit will hydrolyze 1.0 μmole of L-leucine-p-nitroanilide to L-leucine and p-nitroaniline per min at pH 8.0, 25 °C and 3.0 mM substrate concentration.

Physical form

Contains calcium acetate

Preparation Note

Reconstitute in 20 mM tricine, pH 8.0, with 0.05% bovine serum albumin. Dilute the enzyme with the reconstitution buffer to 0.15-0.3 U/mL for a working concentration. Solutions should be prepared fresh prior to use.

Other Notes

Endopeptidase contaminant: Not more than: 0.01 U/mg protein (as μmole tyrosine equivalent per min released from casein.)

Safety Information

• Pictograms: GHS08,GHS07
• Signal Word: Danger
• Hazard Statements: H315,H319,H334,H335
• Precautionary Statements: P261 - P264 - P271 - P302 + P352 - P304 + P340 + P312 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Target Organs: Respiratory system
• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves