The Preparation and Characterization of Cell Walls and the Preparation of Flagella of Vibrio fetus

Abstract

SUMMARYMethods are described for the preparation of cell-wall material and flagella from Vibrio fetus. In common with certain other Gram-negative organisms V. fetus apparently possesses a three-layer wall which, it is proposed, consists of an outer lipoprotein layer, a middle lipo-polysaccharide layer, and an inner mucopeptide layer. These layers were studied chemically and by electron microscopy after various extraction and enzymic degradation procedures with whole-wall material. None of the layers of the wall was found to possess a microscopically discernible substructure, although the extracted lipopolysaccharide agglomerated into a characteristic stringy array. The probable function, contribution to cellular rigidity, and composition of the wall layers is discussed.

Biology of the chemoheterotrophic spirilla.

“…Also, in contrast to most members of Aquaspirillum, the flagella ofBdellovibrio are sheathed. Furthermore, a "polar membrane" such as that described for A. serpens (135) and C. fetus (97,150) does not appear to be present in Bdellovibrio. The DNA base composition of Bdellovibrio is 42 to 43% G+C for some strains and ca.…”

“…These characteristics include: large bipolar fascicles of flagella; a strictly respiratory metabolism; inability to attack carbohydrates; intracellular PHB granules; positive reactions for catalase, oxidase, and phosphatase; lack of tolerance to 3% NaCl or glycine; and a G+C content of 62 to 65%. Furthermore, the occurrence of a "polar membrane" was demonstrated in thin sections; this structure has been found mainly in Aquaspirillum and other helical bacteria (75,76,97,135,149,150). However, it has also been observed in the rod-shaped organism Chromatium (135), and a structure similar in certain respects has also been demonstrated in Selenomonas (31).…”

The flagellar filament protein

“…However, for efficient harvesting they have been detached by shaking (Weibull 1948), occasionally with beads (Keeler 1966), or by blending in a food blender (Stocker and Campbell 1959). Purification is achieved by differential centrifugation to first remove the bacterial bodies and then sediment the free flagella; cuts of the flagella pellet have been made to further purify the product (Kobayashi et al 1959).…”

Fine Structure and Isolation of the Hook-Basal Body Complex of Flagella from Escherichia coli and Bacillus subtilis

“…Data from Proteus (2,15,33), Ectothiorhodospira (27), and Vibrio (31) suggest the same structure. Thin sections through these organisms show a cell wall structure very similar to E. coli (7,11,19,25,27,29,31,32). The basal body from B. subtilis lacks the top set of rings found on the E. coli basal body.…”