Characterization and mass spectrometry analysis of aminopeptidase N from Pseudomonas putida Lup

• Autorzy: Jankiewicz U. , Swiontek-Brzezinska M. , Gorska E. B. , Kowalczyk P.
• Języki publikacji: EN

Abstrakty

EN

An intracellular aminopeptidase N synthesized by Pseudomonas putida Lup was purified and characterized. The approx. 150-fold purified enzyme showed highest activity against A-β-naphthylamide at pH 7.5 and at temperature 40°C and was 100% thermostable for 240 min at 40°C. P. putida lup aminopeptidase N is a monomer with molecular mass approx. 99 kDa determined by SDS-PAGE and gel permeation chromatography. The enzyme has broad substrate specificity, but is the most active against protein substrates with N-terminal alanine and arginine. The activity of P. putida Lup aminopeptidase N is strongly inhibited in the presence of specific metallopeptidase inhibitors and is partly recovered in the presence of Zn²⁺ and Co²⁺ ions. Co²⁺, Mg²⁺ and Ca²⁺ ions increased the activity of the enzyme. Moreover, the enzyme was inhibited by inhibitors of cysteine enzymes. Analysis of fragments of the amino acid sequence of the purified enzyme demonstrated high similarity to PepN of Pseudomonas putida GB-1.

Słowa kluczowe

EN

mass spectrometry analysis; aminopeptidase-N; Pseudomonas putida; enzyme; purification

• Wydawca: -
• Czasopismo: Polish Journal of Microbiology
• Rocznik: 2013
• Tom: 62
• Numer: 4
• Opis fizyczny: p.337-343,fig.,ref.

Twórcy

autor

Jankiewicz U.

• Department of Biochemistry, Warsaw University of Life Sciences – SGGW, Warsaw, Poland

autor

Swiontek-Brzezinska M.

• Department of Environmental Microbiology and Biotechnology, Institute of Ecology and Environmental Protection, Nicolaus Copernicus University, Torun, Poland

autor

Gorska E. B.

• Autonomous Department of Microbial Biology, Warsaw University of Life Sciences – SGGW, Warsaw, Poland

autor

Kowalczyk P.

• Autonomous Department of Microbial Biology, Warsaw University of Life Sciences – SGGW, Warsaw, Poland

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