Aims: The goal of this study was to characterize biopolymers from two modes of the Herbaspirillum lusitanum P6-12 growth: planktonic, in which cells are free swimming, and biofilm life style, in which the cells are sessile. Methods and Results: Differences in biopolymers composition from planktonic and biofilm cells of H. lusitanum strain P6-12 were analysed using Fourier transform infrared spectroscopy (FTIR), sodium dodecyl sulphate-polyacrylamide gel electrophoresis, gas-liquid chromatography and spectrophotometry. A high degree of polymer separation and purification was achieved by ultracentrifugation, and column chromatography allowed us to identify the chemical differences between biopolymers from biofilm and planktonic H. lusitanum. It was shown that planktonic cells of H. lusitanum P6-12 when cultivated in a liquid medium to the end of the exponential phase of growth, produced two high-molecular-weight glycoconjugates (were arbitrarily called CPS-I and CPS-II) of a lipopolysaccharide (LPS) nature and a lipid-polysacharide complex (were arbitrarily called EPS). The EPS, CPS-I, CPS-II had different monosaccharide and lipid compositions. The extracellular polymeric matrix (EPM) produced by the biofilm cells was mostly proteinaceous, with a small amount of carbohydrates (up to 3%). From the biofilm culture medium, a free extracellular polymeric substance (was arbitrarily called fEPS) was obtained that contained proteins and carbohydrates (up to 7%). The cells outside the biofilm had capsules containing high-molecular-weight glycoconjugate (was arbitrarily called CPS FBC) that consisted of carbohydrates (up to 10%), proteins (up to 16%) and lipids (up to 70%). Conclusions: During biofilm formation, the bacteria secreted surface biopolymers that differed from those of the planktonic cells. The heterogeneity of the polysaccharide containing biopolymers of the H. lusitanum P6-12 surface is probably conditioned by their different functions in plant colonization and formation of an efficient symbiosis, as well as in cell adaptation to existence in plant tissues. Significance and Impact of the Study: The results of the study permit a better understanding of the physiological properties of the biopolymers, for example, in plant-microbe interactions.
Linear alkyl sulfates are a major class of surfactants that have large-scale industrial application and thus wide environmental release. These organic pollutants threaten aquatic environments and other environmental compartments. We show the promise of the use of a whole-cell electric sensor in the analysis of low or residual concentrations of sodium dodecyl sulfate (SDS) in aqueous solutions. On the basis of bioinformatic analysis and alkylsulfatase activity determinations, we chose the gram-negative bacterium
Herbaspirillum lusitanum
, strain P6–12, as the sensing element. Strain P6–12 could utilize 0.01–400 mg/L of SDS as a growth substrate. The electric polarizability of cell suspensions changed at all frequencies used (50–3000 kHz). The determination limit of 0.01 mg/L is much lower than the official requirements for the content of SDS in potable and process water (0.5 and 1.0 mg/L, respectively), and the analysis takes about 1–5 min. The promise of
H. lusitanum
P6–12 for use in the remediation of SDS-polluted soils is discussed.
Graphical abstract
Purpose Microbial carbohydrate antigens are targets of the immune systems of hosts. In this context, it is of interest to obtain data that will permit judgment of the degree of heterogeneity, chemical makeup, and localization of the antigenic determinants of the Herbaspirillum surface glycopolymers. Methods A sheep single-chain antibody-fragment phage library (Griffin.1, UK) was used to obtain miniantibodies to the exopolysaccharides (EPS-I and EPS-II), capsular polysaccharides (CPS-I and CPS-II) and lipopolysaccharide (LPS) of Herbaspirillum seropedicae Z78. To infer about the presence or absence of common antigenic determinants in the cell-surface polysaccharides of H. seropedicae Z78, we ran a comparative immunoassay using rabbit polyclonal and phage recombinant antibodies to the surface glycopolymers of H. seropedicae Z78. Results We isolated and purified the exopolysaccharides (EPS-I and EPS-II), capsular polysaccharides (CPS-I and CPS-II), and lipopolysaccharide (LPS) of Herbaspirillum seropedicae Z78. Using rabbit polyclonal antibodies, we found that these cellsurface polysaccharides were of a complex nature. EPS-I, EPS-II, CPS-I, CPS-II, and LPS contained common antigenic determinants. CPS-I, CPS-II, and LPS also contained individual antigenic determinants composed of rhamnose, N-acetyl-D-glucosamine, and N-acetyl-D-galactosamine-sugars responsible for cross-reactions with miniantibodies. Conclusions The anti-LPS miniantibodies were more specific for the core region of the LPS, in which rhamnose was the most abundant sugar, than they were specific for its O portion. The miniantibodies we isolated can be useful reagents not only in basic biochemical research but also in clinical diagnostic and therapeutic applications.
Diazotrophic endobionts Herbaspirillum spp. were studied in respect to the structural peculiarities of the lipopolysaccharides (LPS), O-specific polysaccharides (OPS) structure, antigenic composition and genetics.
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