TY - JOUR
T1 - Purification and characterization of an extracellular non-aspartyl acid protease (pumAe) from Ustilago maydis
AU - Mercado-Flores, Yuridia
AU - Guerra-Sánchez, Guadalupe
AU - Villa-Tanaca, Lourdes
AU - Hernández-Rodríguez, César
PY - 2003/11/1
Y1 - 2003/11/1
N2 - The proteinase pumAe was purified to homogeneity from haploid U. maydis FB1 growing on acid mineral medium. The purification procedure consisted of ammonium sulfate fractionation and gel filtration chromatography, resulting in a 7.7% recovery and a 15.1-fold increase in specific activity. The molecular weight of the enzyme was estimated to be 72 kDa and 74 kDa by gel filtration chromatography and SDS-PAGE, respectively. Enzymatic activity was optimal at pH 4.0 and at 45°C toward hemoglobin, and the pI was determined to be 5.5. The effects of six protease inhibitors on pumAe were tested, and no inhibitory effect was observed. The pure enzyme degraded gelatin and albumin, but casein and collagen were not degraded. The Km value was 3.5 μM, and the Vmax value was 11430 μmol h-1 mg-1 for Suc-R-P-F-H-L-L-V-Y-MCA.
AB - The proteinase pumAe was purified to homogeneity from haploid U. maydis FB1 growing on acid mineral medium. The purification procedure consisted of ammonium sulfate fractionation and gel filtration chromatography, resulting in a 7.7% recovery and a 15.1-fold increase in specific activity. The molecular weight of the enzyme was estimated to be 72 kDa and 74 kDa by gel filtration chromatography and SDS-PAGE, respectively. Enzymatic activity was optimal at pH 4.0 and at 45°C toward hemoglobin, and the pI was determined to be 5.5. The effects of six protease inhibitors on pumAe were tested, and no inhibitory effect was observed. The pure enzyme degraded gelatin and albumin, but casein and collagen were not degraded. The Km value was 3.5 μM, and the Vmax value was 11430 μmol h-1 mg-1 for Suc-R-P-F-H-L-L-V-Y-MCA.
UR - http://www.scopus.com/inward/record.url?scp=0141669018&partnerID=8YFLogxK
U2 - 10.1007/s00284-003-4047-z
DO - 10.1007/s00284-003-4047-z
M3 - Artículo
SN - 0343-8651
VL - 47
SP - 408
EP - 411
JO - Current Microbiology
JF - Current Microbiology
IS - 5
ER -