Isolation, fractionation and some properties of polysaccharides produced in a bound form by<i>Azospirillum brasilense</i>and their possible involvement in<i>Azospirillum</i>-wheat root interactions

Коннова, С. А.; Makarov, O. E.; Skvortsov, Igor M.; Игнатов, В. В.

doi:10.1111/j.1574-6968.1994.tb06809.x

Cited by 72 publications

(12 citation statements)

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“…Bacterial growth, isolation of LPS and OPS . A. halopraeferens strain Au4 (IBPPM 221) [ 5 ] was obtained from the microbial culture collection of the Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences (IBPPM RAS, Saratov) and was cultivated under aerobic conditions at 41 °C in a liquid malate medium [ 8 ] supplemented with 0.09 M NaCl. In an analogous manner as described before [ 14 , 33 ] the cells were washed from the capsule and dried, LPS was extracted from the biomass (10 g) by the Westphal procedure [ 41 ] in a yield 7.4%, and OPS was isolated and purified by gel filtration in a yield 39% of the LPS mass.…”

Section: Methodsmentioning

confidence: 99%

“…A lipopolysaccharide (LPS) having an O-specific polysaccharide chain (OPS) called O antigen and capsular polysaccharide (K antigen) of Azospirillum , which is an extracellular form of LPS [ 8 – 9 ], are important for the interaction between bacteria and host plants. The cell-surface polysaccharides of Azospirillum are involved in overcoming of unfavorable conditions, including survival of bacteria under salinity [ 10 – 13 ].…”

Section: Introductionmentioning

confidence: 99%

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Elucidation of a masked repeating structure of the O-specific polysaccharide of the halotolerant soil bacteria Azospirillum halopraeferens Au4

Sigida¹,

Fedonenko²,

Shashkov³

et al. 2016

Beilstein J. Org. Chem.

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SummaryAn O-specific polysaccharide was obtained by mild acid hydrolysis of the lipopolysaccharide isolated by the phenol–water extraction from the halotolerant soil bacteria Azospirillum halopraeferens type strain Au4. The polysaccharide was studied by sugar and methylation analyses, selective cleavages by Smith degradation and solvolysis with trifluoroacetic acid, one- and two-dimensional 1H and 13C NMR spectroscopy. The following masked repeating structure of the O-specific polysaccharide was established: →3)-α-L-Rhap2Me-(1→3)-[β-D-Glcp-(1→4)]-α-D-Fucp-(1→2)-β-D-Xylp-(1→, where non-stoichiometric substituents, an O-methyl group (~45%) and a side-chain glucose residue (~65%), are shown in italics.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Elucidation of a masked repeating structure of the O-specific polysaccharide of the halotolerant soil bacteria Azospirillum halopraeferens Au4

Sigida¹,

Fedonenko²,

Shashkov³

et al. 2016

Beilstein J. Org. Chem.

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“…From the planktonic cells of H. lusitanum P6-12 EPS crude and CPS crude were extracted and characterized. However, Kdo has been found in the repeating units of the CPS of Rhizobium bacteria (Carlson and Yadav 1985), and in the CPS of some strains of Azospirillum (Konnova et al 1994), and E. coli (Whitfield and Roberts 1999). The even-numbered 3-hydroxy fatty acids are obligatory components of enterobacterial LPS (Maitra et al 1986).…”

Section: Discussionmentioning

confidence: 99%

Characterization of biopolymers produced by planktonic and biofilm cells ofHerbaspirillum lusitanumP6‐12

Velichko

Grinev

Fedonenko

2020

J. Appl. Microbiol.

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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.

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“…Under certain conditions, some bacteria can produce LPS extracellularly. The capsular glycans of some bacteria are represented by LPS (Konnova et al 1994;Whitfield and Roberts 1999;Smol'kina et al 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Characterization of the structure of glycopolymers is necessary for understanding their properties and functions, including those operating in the interaction of bacteria with other organisms and with the surroundings. The principal macromolecules implicated in the recognition of symbiotic partners are exopolysaccharide (EPS), capsular polysaccharide (CPS), and lipopolysaccharide (LPS), which determine the antigenic specificity of gram-negative bacteria (Kato et al 1980;Konnova et al 1994;Skvortsov and Ignatov 1998;Whitfield and Roberts 1999;Yirmiya et al 2000;Smol'kina et al 2010). Much information about the structure of these glycopolymers can be acquired not only by the destructive chemical methods but also by immunochemistry, which enables the study of antigenic determinants in vitro, in vivo, and in planta.…”

Section: Introductionmentioning

confidence: 99%

Phage antibodies for the immunochemical characterization of Herbaspirillum seropedicae Z78 glycopolymers

Velichko

Fedonenko

2019

Ann Microbiol

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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.

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Isolation, fractionation and some properties of polysaccharides produced in a bound form byAzospirillum brasilenseand their possible involvement inAzospirillum-wheat root interactions

Cited by 72 publications

References 11 publications

Elucidation of a masked repeating structure of the O-specific polysaccharide of the halotolerant soil bacteria Azospirillum halopraeferens Au4

Elucidation of a masked repeating structure of the O-specific polysaccharide of the halotolerant soil bacteria Azospirillum halopraeferens Au4

Characterization of biopolymers produced by planktonic and biofilm cells ofHerbaspirillum lusitanumP6‐12

Phage antibodies for the immunochemical characterization of Herbaspirillum seropedicae Z78 glycopolymers

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