Extracellular neuraminidase production by a Pasteurella multocida A:3 strain associated with bovine pneumonia.
The properties of an extracellular neuraminidase produced by a Pasteurella multocida A:3 strain that was isolated in a case of bovine pneumonia were examined during growth in a defined medium. This enzyme (isolated from concentrated culture supernatants of P. multocida A:3) was active against N-acetylneuramin lactose, human alpha-1-acid glycoprotein, fetuin, colominic acid, and bovine submaxillary mucin. Enzyme elaboration was correlated with the growth of the organism in a defined medium, with maximum quantities produced in the stationary phase. The enzyme was purified by a combination of ammonium sulfate fractionation, ion exchange on DEAE-Sephacel, and gel filtration on Sephadex G-200. The purified neuraminidase possessed a specific activity of 9.36 mumol of sialic acid released per min per mg of protein against fetuin. The enzyme possessed a pH optimum of 6.0 and a Km of 0.03 mg/ml. The P. multocida A:3 neuraminidase had a molecular weight of approximately 500,000 as estimated by gel filtration. The enzyme was stable at 4 and 37 degrees C for 3 h. Approximately 75% of the neuraminidase activity was lost within 30 min at 50 degrees C. Greater than 90% of the enzyme activity was destroyed within 10 min at temperatures of > or = 65 degrees C. The P. multocida neuraminidase does not appear to be serologically related to the Pasteurella haemolytica A1 neuraminidase since antiserum prepared against the purified P. haemolytica enzyme did not neutralize the P. multocida enzyme.
White
D J
DJ
Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock 79430, USA.
Jolley
W L
WL
Purdy
C W
CW
Straus
D C
DC
D000818:Animals; D000907:Antibodies, Bacterial; D002417:Cattle; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D016978:Mannheimia haemolytica / Q000201:enzymology; D008970:Molecular Weight; D009439:Neuraminidase / Q000145:classification / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D009500:Neutralization Tests; D016979:Pasteurella multocida / Q000145:classification / Q000201:enzymology* / Q000254:growth & development; D012766:Pasteurellosis, Pneumonic / Q000382:microbiology*; D018410:Pneumonia, Bacterial / Q000382:microbiology / Q000662:veterinary*; D013045:Species Specificity; D013379:Substrate Specificity
D000907:Antibodies, Bacterial; D009439:Neuraminidase