Purification and characterization of an antibacterial protein in the skin secretion of rockfish Sebastes schlegeli.
An antibacterial protein in the skin secretion of rockfish (Sebastes schlegeli) was purified by lectin affinity chromatography on Con A-Sepharose and gel filtration on TSKgel G3000SW. The antibacterial protein featured the high molecular mass and selective action against Gram-negative bacteria. The molecular mass of the protein was estimated to be approximately 150 kDa in gel filtration and approximately 75 kDa by SDS-PAGE, suggesting that it is dimeric. The antibacterial principle was an acidic glycoprotein with pI 4.5, 3.4% reducing sugar and 2.8% amino sugar. Its sugar chains had N-type (high mannose-type) oligosaccharide and sialic acid components. It inhibited strongly the growth of Aeromonas salmonicida, Photobacterium damselae and Shewanella putrefaciens with a minimum inhibitory concentration (MIC) of approximately 3 microg/ml, and moderately the growth of Vibrio parahaemolyticus and A. hydrophila with a MIC of 12.5 microg/ml and 25 microg/ml, respectively. The values of the minimum bactericidal concentration were almost equivalent to those of MIC. The potent sensitivity against virulent pathogens such as A. hydrophila, A. salmonicida and P. damselae may contribute considerably to the innate host defense mechanism to combat microbes on the mucosal surfaces of the rockfish.
Nagashima
Y
Y
Department of Food Science and Technology, Tokyo University of Fisheries, Minato, 108-8477, Tokyo, Japan. yujicd@tokyo-u-fish.ac.jp
Kikuchi
N
N
Shimakura
K
K
Shiomi
K
K
D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005399:Fishes; D006023:Glycoproteins / Q000032:analysis; D006090:Gram-Negative Bacteria / Q000187:drug effects; D008970:Molecular Weight; D009077:Mucins / Q000737:chemistry; D009093:Mucus / Q000737:chemistry / Q000378:metabolism*; D012867:Skin / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D006023:Glycoproteins; D009077:Mucins