Purification and characterization of protease produced by Staphylococcus aureus isolated from a diseased chicken.
A protease produced by Staphylococcus aureus, isolated from a chicken suffering from dermatitis, was purified by successive precipitation with ammonium sulfate, ion-exchange chromatography on Q-Sepharose FF, Sp-Sepharose FF and Mono-Q columns. By Mono-Q column chromatography, two proteases (protease 1 and 2) were obtained. The molecular weights of protease 1 and 2 were estimated at 23.1 and 22.7 kDa, respectively, by SDS-polyacrylamide gel electrophoresis. Their isoelectric points were 5.85 and 5.55, respectively, and they possessed antigenic similarity when examined by the immunoblotting. The N-terminal amino acid sequences of both the proteases were identical (RAQYVNQLKNFKIRETQ). The activities of both the proteases were strongly increased by reducing agents such as L-cysteine and sodium thioglycolate. Their activity was inhibited by thiol protease inhibitors, but was not inhibited by metalloprotease or serine protease inhibitors. From the results, it seems likely that these proteases, produced by S. aureus from diseased chickens, might belong to the thiol protease group.
Takeuchi
S
S
Department of Bioscience, Faculty of Biotechnology, Fukui Prefectural University, Matsuoka, Japan. takeuchi@fpu.ac.jp
Kinoshita
T
T
Kaidoh
T
T
Hashizume
N
N
D000595:Amino Acid Sequence; D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D002364:Caseins / Q000737:chemistry; D011232:Chemical Precipitation; D002645:Chickens; D002852:Chromatography, Ion Exchange / Q000662:veterinary; D003545:Cysteine / Q000737:chemistry; D003872:Dermatitis / Q000382:microbiology / Q000662:veterinary; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D007106:Immune Sera / Q000096:biosynthesis; D007525:Isoelectric Focusing / Q000662:veterinary; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011201:Poultry Diseases / Q000382:microbiology*; D011817:Rabbits; D017421:Sequence Analysis; D013211:Staphylococcus aureus / Q000737:chemistry / Q000201:enzymology*; D013864:Thioglycolates / Q000737:chemistry
D002364:Caseins; D007106:Immune Sera; D013864:Thioglycolates; C026018:azocasein; C017487:2-mercaptoacetate; D010450:Endopeptidases; D003545:Cysteine; D000645:Ammonium Sulfate