APPLIED AND ENVIRONMENTAL MICROBIOLOGY, July 1979, P. 169-172 0099-2240/79/07-0169/04$02.00/0
Vol. 38, No. 1
Exohemagglutinins: New Products of Vibrios KUNIO OISHI,'* SOJI YOKOSHIMA,' TETSUO TOMIYAMA,2 AND KO AIDA' Institute ofApplied Microbiology' and University Branch Hospital,2 The University of Tokyo, Bunkyo-ku, Tokyo, Japan Received for publication 8 February 1979
A number of vibrio strains isolated from marine water produced high units of phytohemagglutinin-like agglutinins. Sugar specificity of the hemagglutinins was different from that of the sugar-binding bacterial toxins and that of the sugarbinding pili on the bacterial cell surfaces. Cholera exoenterotoxin specifically binds to ganglioside GM, in animal cell membranes and causes the hydrolysis of nicotinamide adenine dinucleotide and the consequent activation of adenylate cyclase in the cells (2, 9, 17, 27). This toxin has multiple binding sites for the oligosaccharide portion of GM, (1, 6, 24), and the binding induces temperature-dependent redistribution of the receptors on cell surfaces into aggregates and caps (1, 19). In these points, cholera toxin resembles phytohemagglutinin (phytoHA, lectin), although the toxin exhibits no hemagglutinative activity of phytoHA. The toxin consists of two different kinds of subunits, A and B (3, 10, 14, 15, 23), which are easily separable from each other in the course of purification (4). Subunit A has the activity of NADase (17) and the subunit B has the GMjbinding activity (3) responsible for the phytoHA-like characteristics of the toxin. These subunits were considered to be synthesized in-
dependently and assembled simultaneously (18). These data raise questions whether vibrios are able to produce exohemagglutinin (exoHA) different from the subunit B of the toxin which makes hybrids with the subunit A to form the toxin analogs or acts independently as an antagonist against the binding of the toxin to the animal cell membranes. We have examined the occurrence of exoHA producers in vibrio strains and, if any, the specificity of the HAs for the erythrocytes of various aninal species.
Samples of marine water were collected in June, 1977, from a 700-km-long strip of Pacific coast around Tokyo, inoculated to the plates of thiosulfate-citrate-bile-sucrose medium (Eiken Chemical Co., Ltd., Tokyo, Japan) with the ordinary smearing method, and incubated at 27°C for 1 day. The organisms grown were transferred to marine water containing 3% peptone and 0.5% yeast extract (pH 7.0) and cultured aerobically
TABLE 1. Properties of HA-producing vibrios Vibrio
Growth in NaCl 7% 10% + Dc
0%
V. choleraeb V. parahemolyticusb Biotype1 Biotype 2 V. anguillarumb 363 60
+ -
Voges-Pros- HemagglutinAcid from: kauer ating unita Arabinose Sucrose -
+ +
+
D -
D
-
-
-
+
D
+ +
D
Hemolytic
unt"
+
64 512 512
Vol. 38, No. 1
Exohemagglutinins: New Products of Vibrios KUNIO OISHI,'* SOJI YOKOSHIMA,' TETSUO TOMIYAMA,2 AND KO AIDA' Institute ofApplied Microbiology' and University Branch Hospital,2 The University of Tokyo, Bunkyo-ku, Tokyo, Japan Received for publication 8 February 1979
A number of vibrio strains isolated from marine water produced high units of phytohemagglutinin-like agglutinins. Sugar specificity of the hemagglutinins was different from that of the sugar-binding bacterial toxins and that of the sugarbinding pili on the bacterial cell surfaces. Cholera exoenterotoxin specifically binds to ganglioside GM, in animal cell membranes and causes the hydrolysis of nicotinamide adenine dinucleotide and the consequent activation of adenylate cyclase in the cells (2, 9, 17, 27). This toxin has multiple binding sites for the oligosaccharide portion of GM, (1, 6, 24), and the binding induces temperature-dependent redistribution of the receptors on cell surfaces into aggregates and caps (1, 19). In these points, cholera toxin resembles phytohemagglutinin (phytoHA, lectin), although the toxin exhibits no hemagglutinative activity of phytoHA. The toxin consists of two different kinds of subunits, A and B (3, 10, 14, 15, 23), which are easily separable from each other in the course of purification (4). Subunit A has the activity of NADase (17) and the subunit B has the GMjbinding activity (3) responsible for the phytoHA-like characteristics of the toxin. These subunits were considered to be synthesized in-
dependently and assembled simultaneously (18). These data raise questions whether vibrios are able to produce exohemagglutinin (exoHA) different from the subunit B of the toxin which makes hybrids with the subunit A to form the toxin analogs or acts independently as an antagonist against the binding of the toxin to the animal cell membranes. We have examined the occurrence of exoHA producers in vibrio strains and, if any, the specificity of the HAs for the erythrocytes of various aninal species.
Samples of marine water were collected in June, 1977, from a 700-km-long strip of Pacific coast around Tokyo, inoculated to the plates of thiosulfate-citrate-bile-sucrose medium (Eiken Chemical Co., Ltd., Tokyo, Japan) with the ordinary smearing method, and incubated at 27°C for 1 day. The organisms grown were transferred to marine water containing 3% peptone and 0.5% yeast extract (pH 7.0) and cultured aerobically
TABLE 1. Properties of HA-producing vibrios Vibrio
Growth in NaCl 7% 10% + Dc
0%
V. choleraeb V. parahemolyticusb Biotype1 Biotype 2 V. anguillarumb 363 60
+ -
Voges-Pros- HemagglutinAcid from: kauer ating unita Arabinose Sucrose -
+ +
+
D -
D
-
-
-
+
D
+ +
D
Hemolytic
unt"
+
64 512 512
