Purification and characterization of an extracellular protease produced by Pseudomonas fluorescens M3/6.
Pseudomonas fluorescens strain M3/6 was inoculated into reconstituted NDM and incubated at 7 degrees C for 46 d. A significant amount of extracellular protease was produced, mainly during the latter part of the culture's life cycle. The protease was purified using ammonium sulfate fractionation, ion-exchange chromatography, and gel filtration. The isolated protease had activity on azocasein, alpha-, beta-, and kappa-caseins and a plasmin substrate but did not have plasminogen activator activity. The protease had a molecular weight of 45 kDa, an isoelectric point of pH 8.25, a broad temperature and pH range for activity, and was less heat stable in the isolated form than in the cell-free extract.
Kohlmann
K L
KL
Department of Food Science, Purdue University, West Lafayette, IN 47907.
Nielsen
S S
SS
Ladisch
M R
MR
D000818:Animals; D002364:Caseins / Q000378:metabolism; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D005516:Food Microbiology; D006863:Hydrogen-Ion Concentration; D008892:Milk / Q000382:microbiology*; D008970:Molecular Weight; D010958:Plasminogen / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D011551:Pseudomonas fluorescens / Q000201:enzymology* / Q000254:growth & development; D013696:Temperature
D002364:Caseins; D011480:Protease Inhibitors; D010958:Plasminogen; D010450:Endopeptidases