The chemical structure of polysaccharides isolated from the Ochrobactrum rhizosphaerae PR17T.

The bacteria from Ochrobactrum genus are commonly found in the soil and association with the roots of plants. The O-polysaccharide and the glucan were isolated from the Ochrobactrum rhizosphaerae PR17T strain. Purified polysaccharides were analysed using chemical methods and NMR spectroscopy. Sugar and absolute configuration assignment combined with NMR data revealed the chemical structure of the repeating unit of the O.

Pretreatment with Ochrobactrum immobilizes chromium and copper during sludge pyrolysis.

To increase the degree of immobilization of heavy metals subjected to sludge pyrolysis, we investigated the effects of pretreating sludge with Ochrobactrum supplementation on the immobilization of chromium (Cr) and copper (Cu) during sludge pyrolysis. The sequential extraction procedure was used to test the metallic forms of Cr and Cu. The immobilization of Cr and Cu was characterized with X-ray diffraction, scanning electron microscopy, Fourier transform infrared spectroscopy, etc. Results show that: 1) the addition of Ochrobactrum (1-8%) can accelerate the mineralization process in blank sludge and can accelerate the conversion of the oxidizable forms of Cr and Cu into the residual forms subjected to pyrolysis; 2) pretreatment with Ochrobactrum supplementation can inhibit the volatilization of Cr and Cu during sludge pyrolysis, particularly in the case of a high concentration of Cu. Notably, the pretreatment with Ochrobactrum can reduce 20.38-85.09% of the potential ecological risk of Cr and Cu. The pretreatment with Ochrobactrum contributes to the immobilization of Cr and Cu subjected to sludge pyrolysis and thus can prevent pollution of the environment. The results of this study can be used for harmless disposal of municipal sludge.

Ochrobactrum soli sp. nov., Isolated from a Korean Cattle Farm.

A Gram stain negative, motile, non-spore-forming, rod-shaped, strictly aerobic, beige-pigmented bacterium, designated strain BO-7T, was isolated from soil of cattle farm, in Seosan, Republic of Korea. On the basis of 16S rRNA gene sequencing, strain BO-7T clustered with species of the genus Ochrobactrum and appeared closely related to O. haematophilum CCUG 38531T (98.9%), O. daejeonense KCTC 22458T (98.1%), O. rhizosphaerae DSM 19824T (98.1%), O. pituitosum DSM 22207T (98.0%), and O. pecoris DSM 23868T (98.0%). The digital DNA-DNA hybridization and average nucleotide identity between strain BO-7T and the closely related strains were 21.9-39.1%, 78.5-89.5%, respectively, indicating that BO-7T is a novel species of the genus Ochrobactrum. The DNA G + C content of the genomic DNA was 57.1 mol%, and ubiquinone Q-10 was the predominant respiratory quinone. The polar lipids consisted of phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylmonomethyl-ethanolamine, di-phosphatidylglycerol, the major polyamines were spermidine, putrescine, and sym-homospermidine. The major cellular fatty acids (> 5%) were C16:0, C19:0 cycle ω7c, and C18:1ω7c and/or C18:1ω6c (summed feature 8). ANI calculation, digital DNA-DNA hybridization, physiological and biochemical characteristics indicated that strain BO-7T represents a novel species of the genus Ochrobactrum, for which the name Ochrobactrum soli sp. nov. is proposed. The type strain is BO-7T (= KACC 19676T = LMG 30809T).

Structure, gene cluster of the O antigen and biological activity of the lipopolysaccharide from the rhizospheric bacterium Ochrobactrum cytisi IPA7.2.

Lipopolysaccharide (LPS) of Ochrobactrum cytisi IPA7.2, a bacterium isolated from the roots of Solanum tuberosum L., was extracted from dry bacterial cells and chemically characterized. The O-specific polysaccharide was obtained by mild acid hydrolysis of the LPS and studied by sugar analysis and 1H and 13C NMR spectroscopy, including 1H,1H COSY, 1H,1H TOCSY, 1H,1H ROESY, 1H,13C HSQC, and 1H,13C HMBC experiments. The polysaccharide was linear and consisted of trisaccharide repeating units of the following structure: A putative O-antigen gene cluster of O. cytisi IPA7.2 was identified and found to be consistent with the O-specific polysaccharide structure. The LPS of Ochrobactrum cytisi IPA7.2 promoted the growth of potato microplants in vitro.

Influence of Bacterial Physiology on Processing of Selenite, Biogenesis of Nanomaterials and Their Thermodynamic Stability.

We explored how Ochrobactrum sp. MPV1 can convert up to 2.5 mM selenite within 120 h, surviving the challenge posed by high oxyanion concentrations. The data show that thiol-based biotic chemical reaction(s) occur upon bacterial exposure to low selenite concentrations, whereas enzymatic systems account for oxyanion removal when 2 mM oxyanion is exceeded. The selenite bioprocessing produces selenium nanomaterials, whose size and morphology depend on the bacterial physiology. Selenium nanoparticles were always produced by MPV1 cells, featuring an average diameter ranging between 90 and 140 nm, which we conclude constitutes the thermodynamic stability range for these nanostructures. Alternatively, selenium nanorods were observed for bacterial cells exposed to high selenite concentration or under controlled metabolism. Biogenic nanomaterials were enclosed by an organic material in part composed of amphiphilic biomolecules, which could form nanosized structures independently. Bacterial physiology influences the surface charge characterizing the organic material, suggesting its diverse biomolecular composition and its involvement in the tuning of the nanomaterial morphology. Finally, the organic material is in thermodynamic equilibrium with nanomaterials and responsible for their electrosteric stabilization, as changes in the temperature slightly influence the stability of biogenic compared to chemogenic nanomaterials.

Exploring two contrasting surface-active exopolysaccharides from a single strain of Ochrobactrum utilizing different hydrocarbon substrates.

During production and characterization of exopolysaccharides (EPS) of Ochrobactrum pseudintermedium C1, it was observed that an experimental change in the basic hydrocarbon type of substrate for bacterial utilization led to elicitation of different surface-active properties in the EPS produced. In the sugar substrate, it elicited surfactant property, while in oil substrates it elicited emulsifying property, which indicated that the EPS might be different. Consequently, attention was focused on a detailed analysis of this substrate-specific EPS. Utilizing waste sugar, edible, and mineral oil substrates, EPS produced in each situation was characterized. Besides estimating surface activity and thermostability, each substrate-specific EPS was analyzed by Fourier-transform infrared spectroscopy, gas chromatography-mass spectroscopy, 1 H-nuclear magnetic resonance, and matrix-assisted laser desorption/ionization-time of flight mass spectroscopy to find any structural difference. The results were significantly contrasting although the similarity in molecular mass suggested a basic similarity in polysaccharide structure. Morphological differences were also evident both macroscopically and microscopically with scanning electron microscopy. As the surface-active property of EPS was dependent on the substrate utilized, their structural differences might account for it. These diverse surface activities of EPS produced by a single bacterial strain simply by changing the nature of substrate would also augment their bioapplications. Moreover, utilization of waste and easily available substrates should make such applications convenient, ecofriendly, and cost-worthy.

New Ansamycins from the Deep-Sea-Derived Bacterium Ochrobactrum sp. OUCMDZ-2164.

Two new ansamycins, trienomycins H (1) and I (2), together with the known trienomycinol (3), were isolated from the fermentation broth of the deep-sea-derived bacterium Ochrobactrum sp. OUCMDZ-2164. Their structures, including their absolute configurations, were elucidated based on spectroscopic analyses, ECD spectra, and Marfey's method. Compound 1 exhibited cytotoxic effects on A549 and K562 cell lines with IC50 values of 15 and 23 µM, respectively.

A Novel ZnONPs/PVA-Functionalized Biomaterials for Bacterial Cells Immobilization and its Strengthening Effects on Quinoline Biodegradation.

A novel bacterial cells immobilized carrier (ZnONPs/PVA), polyvinyl alcohol (PVA) composites decorated with ZnO nanoparticles (ZnO NPs), was prepared and used for immobilization of the strain Ochrobactrum sp. LC-1, and subsequently for quinoline degrading in water. Characterization of ZnONPs/PVA by using X-ray diffractometer and scanning electron microscopy demonstrated that ZnO NPs were coated on the surface of PVA cubes evenly and the bacterium grew well on the ZnONPs/PVA. Quinoline biodegradation results showed that the degradation effect of quinoline by ZnONPs/PVA immobilized cells was superior to the free cells significantly. The structure and physical properties of ZnNPs/PVA were maintained steady after the reuse of ZnNPs/PVA for cells immobilization several times. Reusability of the ZnONPs/PVA immobilized cells revealed that the quinoline removal ratio was above 97% within 8 h under the conditions of pH neutral, 37 °C when the initial quinoline concentration was 300 mg/L.

Ochrobactrum sp. MPV1 from a dump of roasted pyrites can be exploited as bacterial catalyst for the biogenesis of selenium and tellurium nanoparticles.

BACKGROUND: Bacteria have developed different mechanisms for the transformation of metalloid oxyanions to non-toxic chemical forms. A number of bacterial isolates so far obtained in axenic culture has shown the ability to bioreduce selenite and tellurite to the elemental state in different conditions along with the formation of nanoparticles-both inside and outside the cells-characterized by a variety of morphological features. This reductive process can be considered of major importance for two reasons: firstly, toxic and soluble (i.e. bioavailable) compounds such as selenite and tellurite are converted to a less toxic chemical forms (i.e. zero valent state); secondly, chalcogen nanoparticles have attracted great interest due to their photoelectric and semiconducting properties. In addition, their exploitation as antimicrobial agents is currently becoming an area of intensive research in medical sciences. RESULTS: In the present study, the bacterial strain Ochrobactrum sp. MPV1, isolated from a dump of roasted arsenopyrites as residues of a formerly sulfuric acid production near Scarlino (Tuscany, Italy) was analyzed for its capability of efficaciously bioreducing the chalcogen oxyanions selenite (SeO32-) and tellurite (TeO32-) to their respective elemental forms (Se0 and Te0) in aerobic conditions, with generation of Se- and Te-nanoparticles (Se- and TeNPs). The isolate could bioconvert 2 mM SeO32- and 0.5 mM TeO32- to the corresponding Se0 and Te0 in 48 and 120 h, respectively. The intracellular accumulation of nanomaterials was demonstrated through electron microscopy. Moreover, several analyses were performed to shed light on the mechanisms involved in SeO32- and TeO32- bioreduction to their elemental states. Results obtained suggested that these oxyanions are bioconverted through two different mechanisms in Ochrobactrum sp. MPV1. Glutathione (GSH) seemed to play a key role in SeO32- bioreduction, while TeO32- bioconversion could be ascribed to the catalytic activity of intracellular NADH-dependent oxidoreductases. The organic coating surrounding biogenic Se- and TeNPs was also characterized through Fourier-transform infrared spectroscopy. This analysis revealed interesting differences among the NPs produced by Ochrobactrum sp. MPV1 and suggested a possible different role of phospholipids and proteins in both biosynthesis and stabilization of such chalcogen-NPs. CONCLUSIONS: In conclusion, Ochrobactrum sp. MPV1 has demonstrated to be an ideal candidate for the bioconversion of toxic oxyanions such as selenite and tellurite to their respective elemental forms, producing intracellular Se- and TeNPs possibly exploitable in biomedical and industrial applications.

Living and Conducting: Coating Individual Bacterial Cells with In†Situ Formed Polypyrrole.

Coating individual bacterial cells with conjugated polymers to endow them with more functionalities is highly desirable. Here, we developed an in situ polymerization method to coat polypyrrole on the surface of individual Shewanella oneidensis MR-1, Escherichia coli, Ochrobacterium anthropic or Streptococcus thermophilus. All of these as-coated cells from different bacterial species displayed enhanced conductivities without affecting viability, suggesting the generality of our coating method. Because of their excellent conductivity, we employed polypyrrole-coated Shewanella oneidensis MR-1 as an anode in microbial fuel cells (MFCs) and found that not only direct contact-based extracellular electron transfer is dramatically enhanced, but also the viability of bacterial cells in MFCs is improved. Our results indicate that coating individual bacteria with conjugated polymers could be a promising strategy to enhance their performance or enrich them with more functionalities.

Structure of the O-specific polysaccharide from the legume endosymbiotic bacterium Ochrobactrum cytisi strain ESC1(T).

The O-specific polysaccharide was obtained from the lipopolysaccharide of the legume-endosymbiotic bacterium Ochrobactrum cytisi strain ESC1(T) and studied by chemical analyses and 1D and 2D NMR spectroscopy. The polysaccharide was found to have a disaccharide repeating unit containing α-d-fucose and ß-N-acetyl-d-galactosamine residues connected via (1→3)-glycosidic bonds, resulting in the following structure: →3)-α-d-Fucp-(1→3)-ß-d-GalpNAc-(1→ The d-GalpNAc residue was nonstoichiometrically substituted with a 4-O-methyl group (∼10%) or with a 4,6-O-(1-carboxy)-ethylidene residue (pyruvyl group) (∼10%).

The synergism of temperature, pH and growth phases on heavy metal biosorption by two environmental isolates.

In real environmental applications, such as heavy metal bioremediation, microorganisms are generally not kept at their optimum growth conditions; therefore, it is imperative to investigate their heavy metal removal performance under diverse environmental conditions. The present study aims to investigate the effects of pH, temperature and growth phases on the removal of Cu(2+) and Cr(6+) by two environmental isolates identified as Ochrobactrum intermedium LBr and Cupriavidus metallidurans CH34. Results showed that cells in logarithmic phase presented better biosorption capacity than cells in stationary phase for both isolates. The Cr(6+) metal was removed more efficiently by live O. intermedium LBr than dead cells; while dead C. metallidurans CH34 biosorbed better than live ones. It was also found that the pH and temperature affected the biosorption capacity. The optimum temperatures were determined to be 37°C and 27°C, and the optimum pH values were 6 and 7 for O. intermedium LBr and C. metallidurans CH34, respectively. Additionally, both microorganisms preferentially adsorbed Cu(2+) in Cu(2+)/Cr(6+) mixtures. The main mechanism of adsorption was determined to be through carboxylic, hydroxyl, and amino functional groups.

Highly sensitive, highly specific whole-cell bioreporters for the detection of chromate in environmental samples.

Microbial bioreporters offer excellent potentialities for the detection of the bioavailable portion of pollutants in contaminated environments, which currently cannot be easily measured. This paper describes the construction and evaluation of two microbial bioreporters designed to detect the bioavailable chromate in contaminated water samples. The developed bioreporters are based on the expression of gfp under the control of the chr promoter and the chrB regulator gene of TnOtChr determinant from Ochrobactrum tritici 5bvl1. pCHRGFP1 Escherichia coli reporter proved to be specific and sensitive, with minimum detectable concentration of 100 nM chromate and did not react with other heavy metals or chemical compounds analysed. In order to have a bioreporter able to be used under different environmental toxics, O. tritici type strain was also engineered to fluoresce in the presence of micromolar levels of chromate and showed to be as specific as the first reporter. Their applicability on environmental samples (spiked Portuguese river water) was also demonstrated using either freshly grown or cryo-preserved cells, a treatment which constitutes an operational advantage. These reporter strains can provide on-demand usability in the field and in a near future may become a powerful tool in identification of chromate-contaminated sites.

[Genotypic and phenotypic similarity between bacteria of Brucella genus and Rhizobiaceae family as a basis for reconsideration of Brucella taxonomy].

Accumulated data on assessment of genome of bacteria from Brucella genus and Rhizobiaceae family (results of sequencing, DNA-rRNA hybridization, 16S rRNA gene sequencing etc.) as well as their phenotypic characteristics (first of all, composition of cell fatty acids) were summarized. Data point to phylogenetic proximity of these bacteria and possibility to unite them in one Rhizobiaceae family together with the closest relatives of Brucella--first of all, with bacteria from Ochrobactrum genus). This seems to be more objective than recreation of Brucellaceae family (Rhizobiales order) with genera Brucella, Ochrobactrum and, possibly, others.

Ochrobactrum haematophilum sp. nov. and Ochrobactrum pseudogrignonense sp. nov., isolated from human clinical specimens.

Three Gram-negative, rod-shaped, non-spore-forming bacteria were isolated from clinical specimens between 1992 and 2000. On the basis of 16S rRNA gene sequence similarities, these strains (CCUG 30717T, CCUG 43892 and CCUG 38531T) were shown to belong to the Alphaproteobacteria, most closely related to Ochrobactrum grignonense (99.0 and 98.2% similarity to the type strain). Chemotaxonomic data (major ubiquinone Q-10; major polyamines spermidine, sym-homospermidine and putrescine; major polar lipids phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol and phosphatidylcholine; major fatty acids C18:1omega7c and C19:0 cyclo omega8c) supported the affiliation of the isolates to the genus Ochrobactrum. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of the isolates from described Ochrobactrum species. Isolates CCUG 30717T and CCUG 43892 were closely related on the basis of DNA-DNA reassociation experiments and therefore represent one novel species, for which the name Ochrobactrum pseudogrignonense sp. nov. is proposed, with the type strain CCUG 30717T (=CIP 109451T). Isolate CCUG 38531T was different from these strains and also from other Ochrobactrum species. For this strain, the name Ochrobactrum haematophilum sp. nov. is proposed, with the type strain CCUG 38531T (=CIP 109452T).

Ochrobactrum pseudintermedium sp. nov., a novel member of the family Brucellaceae, isolated from human clinical samples.

Three novel Gram-negative, non-fermenting aerobic bacilli were isolated from human clinical samples. They shared more than 99.8 % of the 16S rRNA gene nucleotide positions. The strains were related to Ochrobactrum intermedium with about 97.48 % 16S rRNA gene sequence similarity. In 16S rRNA gene-, dnaK- and rpoB-based phylogenies, the strains were grouped in a lineage that was distinct from other Ochrobactrum species in the family Brucellaceae. Fatty acid composition, polar lipids, quinone system, DNA-DNA relatedness, genome organization, and physiological and biochemical data differentiated these isolates from recognized species of the genus Ochrobactrum. The three clinical strains therefore represent a novel species within the genus Ochrobactrum, for which the name Ochrobactrum pseudintermedium sp. nov., is proposed. The type strain is ADV31(T) (=CIP 109116(T)=DSM 17490(T)). The DNA G+C content of strain ADV31(T) was 54.5 mol%.

Description of Pseudochrobactrum gen. nov., with the two species Pseudochrobactrum asaccharolyticum sp. nov. and Pseudochrobactrum saccharolyticum sp. nov.

Two Gram-negative, rod-shaped, oxidase-positive, non-spore-forming, non-motile bacteria (CCUG 46016(T) and CCUG 33852(T)), isolated from a knee aspirate of a 66-year-old man and an industrial glue, respectively, were studied for their taxonomic position. On the basis of chemotaxonomic data [i.e. major ubiquinone (Q-10), major polar lipids (phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine) and major fatty acids (C(18 : 1)omega7c and C(19 : 0) cyclo omega8c)] and 16S rRNA gene sequence similarity, both strains belong to the Alphaproteobacteria. The presence of spermidine and putrescine as the predominant polyamines in CCUG 46016(T) were in agreement with its phylogenetic affiliation in the vicinity of the genus Ochrobactrum. 16S rRNA gene sequence similarities between both strains and established species within the genera Bartonella, Ochrobactrum and Brucella were less than 95 %. Although both organisms showed highest 16S rRNA gene sequence similarity to members of Brucella, phenotypic features (including chemotaxonomic features) were more like those of members of the genus Ochrobactrum. Sequence comparison of the recA genes confirmed the separate phylogenetic position of the two strains. On the basis of DNA-DNA pairing results and physiological and biochemical data, the two strains can be clearly differentiated from each other and from all known Ochrobactrum species. It is evident that these organisms represent two novel species in a new genus, Pseudochrobactrum gen. nov., for which the names Pseudochrobactrum asaccharolyticum sp. nov. (the type species, type strain CCUG 46016(T)=CIP 108977(T)) and Pseudochrobactrum saccharolyticum sp. nov. (type strain CCUG 33852(T)=CIP 108976(T)) are proposed.