Aeromonas spp. can secrete Escherichia coli alkaline phosphatase into the culture supernatant, and its release requires a functional general secretion pathway.
Aerolysin is a channel-forming protein secreted by Aeromonas hydrophila. To determine if regions of aerolysin could direct the secretion of another protein, portions of aerA were fused to phoA, the Escherichia coli alkaline phosphatase gene and cloned into E. coli, Aeromonas salmonicida, and A. hydrophila. We were surprised to find that secretion of the enzyme by both Aeromonas spp. was independent of the aerolysin segments fused to it. The smallest fusion product contained only the signal sequence and two amino acids of aerolysin. The largest had more than 90% of the aerolysin molecule. The fusion proteins were found in the periplasms of E. coli and A. salmonicida grown in LB medium containing glucose, as well as in the shocked cells. Aerolysin itself was secreted by A. salmonicida under these conditions. In contrast, when A. salmonicida containing any of the fused genes was grown in LB medium without glucose, most of the alkaline phosphatase activity was extracellular, whereas beta-lactamase remained in its normal periplasmic location. Similar results were obtained with A. hydrophila. The change in location of the enzyme in A. salmonicida appeared to be related to the pH of the growth medium. A. salmonicida and A. hydrophila also secreted native E. coli alkaline phosphatase, but A. hydrophila strains with mutations in the general secretion pathway were unable to release the enzyme. We conclude that the Aeromonas secretion system can recognize the E. coli enzyme as an extracellular protein and direct it outside the cell.
Wong
K R
KR
Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
Buckley
J T
JT
D000333:Aeromonas / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D016980:Aeromonas hydrophila / Q000235:genetics / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000096:biosynthesis* / Q000302:isolation & purification; D001345:Autoradiography; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005798:Genes, Bacterial; D005947:Glucose / Q000378:metabolism; D015151:Immunoblotting; D007473:Ion Channels / Q000096:biosynthesis / Q000302:isolation & purification; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D052899:Pore Forming Cytotoxic Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D013462:Sulfur Radioisotopes; D013997:Time Factors
D001427:Bacterial Toxins; D004251:DNA Transposable Elements; D007473:Ion Channels; D052899:Pore Forming Cytotoxic Proteins; D011993:Recombinant Fusion Proteins; D013462:Sulfur Radioisotopes; C032676:aerolysin; D008715:Methionine; D000469:Alkaline Phosphatase; D005947:Glucose