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A9934 Sigma-Aldrich

Aminopeptidase I from Streptomyces griseus

lyophilized powder, ≥200 units/mg protein

Synonym: Leucine Aminopeptidase IV

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Properties

Related Categories 3.4.x.x Peptidases, 3.x.x.x Hydrolases, Application Index, Biochemicals and Reagents, Enzyme Class Index,
Quality Level   200
form   lyophilized powder
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

Packaging

0.1 mg in serum bottle

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 & Documentation

Safety Information

Personal Protective Equipment 
RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 1
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Documents

Certificate of Analysis (COA)

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Certificate of Origin (COO)

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Protocols & Articles

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