Quantitative Determination of Amines Produced by the Strains of Shigella and Escherichia

(Significance of the bacterial enzymes in ekiri, III)

Abstract

The amount of each of the amines which were produced by each strain of the groups, of Shigella and a strain of E. coli isolated from ekiri patients were measured by using paperchromatography.
Each strain was inoculated into the meat extract broth (pH. 7.2) containing 2 per cent glucose, 100γ/ml pyridoxine and 0.2 er cent amino acid (substrate).
After 16 hours incubation at 37° the cells were harvested by centrifugation, washed, resuspended in acetic acid buffer solution containing amino acid substrate in M/20 concentration, and incubated for 24 hours at 37°. After incubation the mixture was treated with alcohol, centrifuged, nd the precipitated protein was discarded. The supernatant was condensed and was dissolved into 0.1 ml of distilled water, and the total volume was put on a filter paper and was developed chromatographically. The clipped amine spots were eluted into 75 per cent aceton solution and the optical density was measured lorimetrically.
The amount of amine per mg N of the cells was calculated comparing with the standard curve.
Results obtained were as follows:
(1) The amount of histamine was the greatest in all amines produced.
(2) Agmatine was produced by all strains in a considerable amount, especially the Kondo Strain which was isolated from the stool of an ekiri patient.
(3) The amount of cadaverine produced was more than histamine but less than putrescine.
(4) utrescine and ry-aminobutyric acid were produced by all strains and their amounts were next to agmatine.
(5) Tyramine was not produced by all strains.
(6) The relationship between the pathogenesis of ekiri and the amount of amines produced in the intestinal canal by Shigella or Escherichia was discussed.