TY - JOUR
T1 - Purification and partial characterization of a Nocardia brasiliensis extracellular protease
AU - Zlotnik, H.
AU - Schramm, V. L.
AU - Buckley, H. R.
PY - 1984
Y1 - 1984
N2 - Nocardia brasiliensis possess proteolytic activities that can be readily detected in a variety of media. In a modified formulation of a growth medium originally used for Streptomyces aureofaciens, N. brasiliensis was found to secrete proteolytic enzymes, one of which was capable of hydrolyzing casein. This enzyme was purified to homogeneity from cell-free culture filtrates of N. brasiliensis. The purification procedure included ion-exchange chromatography on carboxymethyl-Sepharose, gel filtration of Sephadex G-100, and affinity chromatography, using a hemoglobin-Sepharose resin. The molecular weight of the N. brasiliensis protease was found to be 25,000 by gel filtration and 35,000 by sodium dodecyl sulfate-discontinuous gel electrophoresis. The enzyme is inhibited by o-phenanthroline and 8-hydroxyquinoline-5-sulfonic acid but is not affected by EDTA. Average values for its kinetic parameters were 0.288 μmol of hemoglobin solubilized per min per mg of enzyme for V(max) and 0.76 mM for K(m), using hemoglobin as the substrate.
AB - Nocardia brasiliensis possess proteolytic activities that can be readily detected in a variety of media. In a modified formulation of a growth medium originally used for Streptomyces aureofaciens, N. brasiliensis was found to secrete proteolytic enzymes, one of which was capable of hydrolyzing casein. This enzyme was purified to homogeneity from cell-free culture filtrates of N. brasiliensis. The purification procedure included ion-exchange chromatography on carboxymethyl-Sepharose, gel filtration of Sephadex G-100, and affinity chromatography, using a hemoglobin-Sepharose resin. The molecular weight of the N. brasiliensis protease was found to be 25,000 by gel filtration and 35,000 by sodium dodecyl sulfate-discontinuous gel electrophoresis. The enzyme is inhibited by o-phenanthroline and 8-hydroxyquinoline-5-sulfonic acid but is not affected by EDTA. Average values for its kinetic parameters were 0.288 μmol of hemoglobin solubilized per min per mg of enzyme for V(max) and 0.76 mM for K(m), using hemoglobin as the substrate.
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U2 - 10.1128/jb.157.2.627-631.1984
DO - 10.1128/jb.157.2.627-631.1984
M3 - Article
C2 - 6363390
AN - SCOPUS:0021351157
SN - 0021-9193
VL - 157
SP - 627
EP - 631
JO - Journal of Bacteriology
JF - Journal of Bacteriology
IS - 2
ER -