Isolation and characterization of two immunochemically distinct alkaline phosphatases from Pseudomonas aeruginosa.
We have isolated two alkaline phosphatases (H-AP and L-AP, for high and low molecular mass, respectively) from Pseudomonas aeruginosa PA01. These two enzymes were found to differ in mobility on sodium dodecyl sulphate polyacrylamide gels (H-AP, M(r) = 51,000 and L-AP, M(r) = 39,500), amino-terminal amino acid sequence and did not cross-react. Both enzymes were active as phosphomonoesterases while only L-AP demonstrated any phosphodiesterase activity. Both enzymes were purified from P. aeruginosa grown in phosphate limiting conditions using the same protocol and were identified in both periplasmic and extracellular locations. A low level of H-AP was produced constitutively whereas L-AP was produced only after induction by reduced phosphate concentration in the growth medium. An L-AP-like enzyme has been previously described, however, this is the first report of a second P. aeruginosa alkaline phosphatase.
Tan
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AS
Department of Microbiology, University of Manitoba, Winnipeg, Canada.
Worobec
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EA
D000469:Alkaline Phosphatase / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D001693:Biological Transport, Active; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D015870:Gene Expression; D007120:Immunochemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010710:Phosphates / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D010710:Phosphates; D000469:Alkaline Phosphatase