Preliminary studies on the bacterial substance extracted from oral gram-negative bacteria]

In previous papers, Miura (Kuwahara), Fujimaki and Miura (Kuwahara) reported that the metabolic pathways of the genus Neisseria isolated from the oral cavity showed different patterns under various conditions. On the other hand, there is little reported about the bacterial substance of this genus.In the present paper, we report a preliminary attempt to extract the crude bacterial substance.The extraction of bacterial substance with carried out with the method of Takeda et al. using 0.3% diluted acetic acid solution.The biochemical properties of this extracted substance was based upon the following three experiments.1) Analysis of nitrogen, phosphate, reduced sugar and other reactions for the major component of this material is as follows:N: 3.0-3.5%, P (total P): 1.0-1.5%, reduced sugar (as glucose): 20-21%Biuretu0027s reaction: ++, Molischu0027s reaction: ++Elson-Morganu0027s reaction: +2) For the protein estimation, the ultraviolet absorption from 220 mμ to 340 mμ was studied using a Beckman DU2 spectrophotometer.The results showed a gentle absorption band at about 280 mμ for a 0.01 N NaOH solution of this material.3) Electrophoretic analysis was also made according to the method of zone-thin-layer procedure by agar gel.a) After zone electrophoresis, a 1 cm piece of the agar block was cut off and the protein fraction at 660 mμ was determined colorimetrically using Folin-phenol reagent. Two peaks were observed toward the cathode and one peak toward the anode.b) On the other hand, the thin-layer method showed only one migrating band on the anode side using suddan-BB dye.These results seem to indicate a lipopolysaccharide protein complex as the major component of this bacterial substance. Further details will be published later.