[Atypical R-S dissociation in Azospirillum brasilense]. / Netipichnyi kharakter R-S dissotsiatsii Azospirillum brasilense.

It was found that atypical R-S dissociation in the type strain A. brasilense Sp7 is not accompanied by drastic changes in the carbohydrate moieties of bacterial lipopolysaccharides but is rather due to different contributions of two O-specific polysaccharides (found in both R and S dissociants) to the age-dependent architectonics of the cell surface.

Trace cobalt speciation in bacteria and at enzymic active sites using emission Mössbauer spectroscopy.

57Co emission Mössbauer spectroscopy (EMS) allows the chemical state of cobalt, as influenced by its coordination environment, to be monitored in biological samples at its physiological (trace) concentrations. To draw attention to EMS as a valuable tool for speciation of cobalt in biocomplexes, the process of cobalt(II) metabolism in cells of the plant growth-promoting rhizobacterium Azospirillum brasilense Sp245 was investigated using EMS of 57CoII-doped bacterial cells. EMS measurements also showed 57CoII-activated glutamine synthetase (GS, a key enzyme of nitrogen metabolism, isolated from this bacterium) to have two different cobalt(II) forms at its active sites, in agreement with data available on other bacterial GSs. Chemical after-effects following electron capture by the nucleus of the parent 57CoII during the 57Co-->57Fe transition, which contribute to the formation of a stabilised daughter 57FeIII component along with the nucleogenic 57FeII forms, are also briefly considered.

Structural effects of the Azospirillum lipopolysaccharides in cell suspensions.

The structural influence of Azospirillum lipopolysaccharides (LPS) and lipopolysaccharide-protein complexes (LPPC) on carrot, erythrocyte, and bacterial cell suspensions was explored. The structural potentialities of O-specific polysaccharide fragments of LPS and protein fractions of LPPC were also evaluated. An ability to induce the formation of three kinds of structures in the cell suspensions was revealed depending on the chemical composition of the preparations used. The first and the second ones were connected with effects of cell aggregation (a relatively fast process) and agglutination (a relatively slow process). The third one resulted in phase separation of erythrocyte suspensions (a medium-speed process), with segregating the cells to a separate homogeneous liquid phase.

Influence of divalent cations on the catalytic properties and secondary structure of unadenylylated glutamine synthetase from Azospirillum brasilense.

Fully unadenylylated glutamine synthetase (GS) from the endophytic bacterium Azospirillum brasilense Sp245 was isolated and purified. The enzyme was electrophoretically homogeneous and contained strongly bound metal ions, which could not be removed by dialysis. Mn2+, Mg2+, and Co2+ were found to be effective in supporting biosynthetic activity of the A. brasilense GS. Some kinetic properties of Mn2+-activated and Mg2+-activated unadenylylated GS were characterized. Circular dichroism analysis of the enzyme showed that the A. brasilense GS is a highly structured protein: 59% of its residues form alpha-helices and 13% beta-strands. Removal of the metal ions from the A. brasilense GS by treatment with EDTA resulted in alterations in the enzyme secondary structure.

Involvement of a 120-MDa plasmid of Azospirillum brasilense Sp245 in the production of lipopolysaccharides.

Using Omegon-Km mutagenesis, six Azospirillum brasilense Sp245 mutant derivatives lacking the capability to synthesize either one of the two major O-specific polysaccharides (O-PSs) were constructed in vivo. In all of the Lps mutants obtained, single Omegon-Km insertions were shown to be located on an indigenous plasmid DNA with molecular weight 120 MDa (p120). Physical and immunochemical analyses revealed two p120 loci coding for O-PSI and two p120 loci involved in the production of O-PSII. One of the lps loci from both groups was also shown to act in the production of Calcofluor-binding polysaccharides. It was demonstrated that two Sp245 plasmid bands with apparent molecular weights of 120 and 130 MDa (as visualized by analytical gel electrophoreses) seem to be the two topological forms of the same plasmid species (p120). Transfer properties of p120 were also examined.