TY - JOUR
T1 - Purification and characterization of aminopeptidase (pumAPE) from Ustilago maydis
AU - Mercado-Flores, Yuridia
AU - Noriega-Reyes, Yamilet
AU - Ramírez-Zavala, Bernardo
AU - Hernández-Rodríguez, César
AU - Villa-Tanaca, Lourdes
N1 - Funding Information:
Y. Mercado-Flores, B. Ramı́rez-Zavala, and Y. Noriega-Reyes are fellows from CONACyT and PIFI, IPN. C. Hernández-Rodrı́guez and L. Villa-Tanaca received COFAA, and EDD, IPN supports. This work was supported by Grant CGPI 20030462, IPN, Mexico. L. Villa-Tanaca was hired by “Programa Institucional de Contratación de Personal Académico de Excelencia para el Reforzamiento del Posgrado, IPN”. This work was awarded the “Premio Nacional de Ciencia y Tecnologı́a en Alimentos 2003”
PY - 2004/5/15
Y1 - 2004/5/15
N2 - The aminopeptidase pumAPE was purified from the haploid fungus Ustilago maydis FB1 strain. The purification procedure consisted of ammonium sulfate fractionation and three chromatographic steps, which included anion-exchange, hydrophobic interaction, and gel filtration chromatography, resulting in a 23% recovery. The molecular mass of the dimeric enzyme was estimated to be 110 kDa and 58 kDa by gel filtration chromatography and SDS-PAGE, respectively. Enzymatic activity was optimal at pH 7.0 and at 35°C toward Lys-pNA and the pI was determined to be 5.1. The enzyme was inhibited by EDTA-Na2, 1,10- phenanthroline, bestantin, PMSF and several divalent cations (Cu 2+, Hg2+ and Zn2+). The aminopeptidase showed a preference for lysine and arginine in the N-position. The Km value was 54.4 μM and the Vmax value was 408 μmolmin -1mg-1 for Lys-pNA.
AB - The aminopeptidase pumAPE was purified from the haploid fungus Ustilago maydis FB1 strain. The purification procedure consisted of ammonium sulfate fractionation and three chromatographic steps, which included anion-exchange, hydrophobic interaction, and gel filtration chromatography, resulting in a 23% recovery. The molecular mass of the dimeric enzyme was estimated to be 110 kDa and 58 kDa by gel filtration chromatography and SDS-PAGE, respectively. Enzymatic activity was optimal at pH 7.0 and at 35°C toward Lys-pNA and the pI was determined to be 5.1. The enzyme was inhibited by EDTA-Na2, 1,10- phenanthroline, bestantin, PMSF and several divalent cations (Cu 2+, Hg2+ and Zn2+). The aminopeptidase showed a preference for lysine and arginine in the N-position. The Km value was 54.4 μM and the Vmax value was 408 μmolmin -1mg-1 for Lys-pNA.
KW - Aminopetidase pumAPE
KW - Purification
KW - Ustilago maydis
UR - http://www.scopus.com/inward/record.url?scp=2342640068&partnerID=8YFLogxK
U2 - 10.1016/j.femsle.2004.03.035
DO - 10.1016/j.femsle.2004.03.035
M3 - Artículo
SN - 0378-1097
VL - 234
SP - 247
EP - 253
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
IS - 2
ER -