Purification and characterization of x-prolyl-dipeptidyl aminopeptidase from Lactococcus lactis subsp. cremoris NRRL 634
| Autor: | Teresa Cruz y Victoria, Alfredo E. Pérez-Guzmán, Ramón Cruz-Camarillo, Humberto Hernández-Sánchez |
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| Rok vydání: | 2006 |
| Předmět: |
chemistry.chemical_classification
biology Physiology Lactococcus lactis subsp cremoris Lactococcus lactis General Medicine biology.organism_classification Applied Microbiology and Biotechnology Aminopeptidase Michaelis–Menten kinetics Endopeptidase Enzyme assay Enzyme chemistry Biochemistry biology.protein Polyacrylamide gel electrophoresis Biotechnology |
| Zdroj: | World Journal of Microbiology and Biotechnology. 22:953-958 |
| ISSN: | 1573-0972 0959-3993 |
| DOI: | 10.1007/s11274-006-9140-6 |
| Popis: | An X-prolyl-dipeptidylaminopep tidase (Pep-XP) was purified from the crude intracellular extract of Lactococcus lactis subsp. cremoris NRRL 634 by ion exchange and gel filtration chromatographies. The enzyme was purified 80-fold with a recovery of 6%, and appeared as a single band with a molecular weight of about 80 kDa on polyacrylamide gel electrophoresis with sodium dodecyl sulphate (SDS-PAGE). The peptidase showed its maximal activity on arginyl-proline-p-nitroanilide at pH 7.0 and at a temperature of 45 °C, although there was a good activity of Pep-XP in the pH range of 5.5–7.0 and temperatures between 40 and 50 °C. The Michaelis constant (Km) and the maximum reaction velocity (Vmax) values were 0.92 mM and 7.9 U/mg protein min, respectively. The activity of Pep-XP was completely inhibited by phenylmethanesulphonyl fluoride, an inhibitor of serine peptidases, and metal chelators had little effect on enzyme activity. The purified enzyme hydrolyzed synthetic substrates whose structure is X-Pro-Y like Lys-Pro-pNA, but did not hydrolyse Pro-pNA or azocasein, showing that the enzyme did not have aminopeptidase or endopeptidase activities. |
| Databáze: | OpenAIRE |
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