Edman degradation and protein sequencing: Pfu Aminopeptidase I
Edman degradation is a commonly used method for protein sequencing via N-terminal amino acid cleavage. Pfu Aminopeptidase I is a thermostable exo-type aminopeptidase isolated from Pyrococcus furiosus. It is expressed as a recombinant protein and liberates N-terminal amino acids from proteins and peptides. While Pfu Aminopeptidase I has a wide range of substrate specificities, it does not hydrolyze peptide bonds at the α-amino group of proline (X-Pro). Enzyme activity is significantly increased in the presence of Co2+.
- Liberates the N-terminal amino acids up to X-Pro from proteins and peptides
Recombinant Escherichia coli encoding the Pyrococcus furiosus aminopeptidase I gene
- Molecular weight: 37.483 kDa (calculated), 36–37 kDa (SDS-PAGE)
- Isoelectric point: 4.6–4.65
- Inhibitor: EDTA (completely inhibited at 0.1 mM)
- Optimum pH: 5.5–6.0 (in the absence of Co2+ at 90°C)
5.5–8.0 (in the presence of 20 µM Co2+ at 90°C)
- Optimum temperature: 80°C (pH 6.0 in the absence of Co2+) 95°C (pH 6.0 in the presence of 20 µM Co2+)
- Thermal stability: 65% activity after 4 hours at 90°C (without Co2+ at pH 8.0)
Definition of activity
One unit of enzyme activity corresponds to the amount required to hydrolyze 1 µmol of Leucine-p-nitroanilide at 75°C, pH 8.0 in one minute.
- >16 U/mg protein (without Co2+)
- >80 U/mg protein (in the presence of 20 µM Co2+)
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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