TY - JOUR
T1 - Purification and characterization of a lysine aminopeptidase from Kluyveromyces marxianus
AU - Ramírez-Zavala, Bernardo
AU - Mercado-Flores, Yuridia
AU - Hernández-Rodríguez, César
AU - Villa-Tanaca, Lourdes
N1 - Funding Information:
This study was supported by a research grant from CONACYT (26437-N) and CGPI-IPN (20020679), B. Ramı́rez-Zavala and Y. Mercado-Flores were fellows from CONACYT and PIFI-IPN. L. Villa-Tanaca and C. Hernández-Rodrı́guez received COFAA-IPN and EDD-IPN support. L. Villa-Tanaca was hired by the “Programa de Contratación para el Apoyo a la Investigación y el Posgrado del IPN”.
PY - 2004/6/15
Y1 - 2004/6/15
N2 - A lysine aminopeptidase was purified from the yeast Kluyveromyces marxianus. This enzyme was purified 100-fold from a soluble extract obtained at 100,000g. The purification procedure consisted in fractionated precipitation with ammonium sulfate and five chromatography steps. The native enzyme had a molecular mass of 46 kDa assessed through gel filtration. This aminopeptidase depicted an optimal pH of 7.0 and was stable at a pH range of 4-8, its optimal temperature was 45°C and the enzyme became unstable at temperatures above 55°C. The isoelectric point of the purified enzyme was 4.4. Michaelis constant and Vmax for L-lysine-p-nitroanilide were 0.33 mM and 2.2 mM min-1 per milligram of protein, respectively. The enzyme was strongly inhibited by bestatin, o-phenanthroline and, to a lesser extent, by EDTA, suggesting that this enzyme is a metalloprotease. Our results suggest that the lysine aminopeptidase from Kluyveromyces marxianus might be of biotechnological relevance.
AB - A lysine aminopeptidase was purified from the yeast Kluyveromyces marxianus. This enzyme was purified 100-fold from a soluble extract obtained at 100,000g. The purification procedure consisted in fractionated precipitation with ammonium sulfate and five chromatography steps. The native enzyme had a molecular mass of 46 kDa assessed through gel filtration. This aminopeptidase depicted an optimal pH of 7.0 and was stable at a pH range of 4-8, its optimal temperature was 45°C and the enzyme became unstable at temperatures above 55°C. The isoelectric point of the purified enzyme was 4.4. Michaelis constant and Vmax for L-lysine-p-nitroanilide were 0.33 mM and 2.2 mM min-1 per milligram of protein, respectively. The enzyme was strongly inhibited by bestatin, o-phenanthroline and, to a lesser extent, by EDTA, suggesting that this enzyme is a metalloprotease. Our results suggest that the lysine aminopeptidase from Kluyveromyces marxianus might be of biotechnological relevance.
KW - Aminopeptidase
KW - Kluyveromyces marxianus
KW - Protease
KW - Purification
UR - http://www.scopus.com/inward/record.url?scp=2942613202&partnerID=8YFLogxK
U2 - 10.1016/j.femsle.2004.05.009
DO - 10.1016/j.femsle.2004.05.009
M3 - Artículo
SN - 0378-1097
VL - 235
SP - 369
EP - 375
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
IS - 2
ER -