573 research outputs found

    Highly hydrolytic reuteransucrase from probiotic Lactobacillus reuteri strain ATCC 55730

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    Lactobacillus reuteri strain ATCC 55730 (LB BIO) was isolated as a pure culture from a Reuteri tablet purchased from the BioGaia company. This probiotic strain produces a soluble glucan (reuteran), in which the majority of the linkages are of the α-(1→4) glucosidic type (∼70%). This reuteran also contains α-(1→6)- linked glucosyl units and 4,6-disubstituted α-glucosyl units at the branching points. The LB BIO glucansucrase gene (gtfO) was cloned and expressed in Escherichia coli, and the GTFO enzyme was purified. The recombinant GTFO enzyme and the LB BIO culture supernatants synthesized identical glucan polymers with respect to linkage type and size distribution. GTFO thus is a reuteransucrase, responsible for synthesis of this reuteran polymer in LB BIO. The preference of GTFO for synthesizing α-(1→4) linkages is also evident from the oligosaccharides produced from sucrose with different acceptor substrates, e.g., isopanose from isomaltose. GTFO has a relatively high hydrolysis/transferase activity ratio. Complete conversion of 100 mM sucrose by GTFO nevertheless yielded large amounts of reuteran, although more than 50% of sucrose was converted into glucose. This is only the second example of the isolation and characterization of a reuteransucrase and its reuteran product, both found in different L. reuteri strains. GTFO synthesizes a reuteran with the highest amount of α-(1→4) linkages reported to date

    Efficient screening methods for glucosyltransferase genes in Lactobacillus strains

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    Limited information is available about homopolysaccharide synthesis in the genus Lactobacillus. Using efficient screening techniques, extracellular glucosyltransferase (GTF) enzyme activity, resulting in α-glucan synthesis from sucrose, was detected in various lactobacilli. PCR with degenerate primers based on homologous boxes of known glucosyltransferase (gtf) genes of lactic acid bacteria strains allowed cloning of fragments of 10 putative gtf genes from eight different glucan producing Lactobacillus strains (five Lactobacillus reuteri strains, one Lactobacillus fermentum strain, one Lactobacillus sake strain and one Lactobacillus parabuchneri strain). Sequence analysis revealed that these lactobacilli possess a large variation of (putative) gtf genes, similar to what has been observed for Leuconostoc and Streptococcus strains. Homologs of GTFA of Lb. reuteri 121 (synthesizing reuteran, a unique glucan with α-(1→4) and α-(1→6) glycosidic bonds) were found in three of the four other Lb. reuteri strains tested. The other Lactobacillus GTF fragments showed the highest similarity with GTF enzymes of Leuconostoc spp.

    Palladium(II) complexes of quinolinylaminophosphonates: synthesis, structural characterization, antitumor and antimicrobial activity

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    Three types of palladium(II) halide complexes of quinolinylaminophosphonates have been synthesized and studied. Diethyl and dibutyl [alpha-anilino-(quinolin-2-ylmethyl)]phosphonates (L1, 12) act as N,N-chelate ligands through the quinoline and aniline nitrogens giving complexes cis-[Pd(L1/12)X-2] (X Cl, Br) (1-4). Their 3-substituted analogues [alpha-anilino-(quinolin-3-ylmethyl)]phosphonates (L3, L4) form dihalidopalladium complexes trans-[Pd(L3/L4)(2)X-2] (5-8), with trans N-bonded ligand molecules only through the quinoline nitrogen. Dialkyl [alpha-(quinolin-3-ylamino)-N-benzyl]phosphonates (L5, L6) give tetrahalidodipalladium complexes [Pd-2(L5/L6)(3)X-4] (9-12), containing one bridging and two terminal ligand molecules. The bridging molecule is bonded to the both palladium atoms, one through the quinoline and the other through the aminoquinoline nitrogen, whereas terminal ligand molecules are coordinated each only to one palladium via the quinoline nitrogen. Each palladium ion is also bonded to two halide ions in a trans square-planar fashion. The new complexes were identified and characterized by elemental analyses and by IR, UV-visible, H-1, C-13 and P-31 nuclear magnetic resonance and ESI-mass spectroscopic studies. The crystal structures of complexes 1-4 and 6 were determined by X-ray structure analysis. The antitumor activity of complexes in vitro was investigated on several human tumor cell lines and the highest activity with cell growth inhibitory effects in the low micromolar range was observed for dipalladium complexes 11 and 12 derived from dibutyl ester L6. The antimicrobial properties in vitro of ligands and their complexes were studied using a wide spectrum of bacterial and fungal strains. No specific activity was noted. Only ligands L3 and L4 and tetrahalidodipalladium complexes 9 and 11 show poor activities against some Gram positive bacteria

    Structure-function relationships of glucansucrase and fructansucrase enzymes from lactic acid bacteria

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    Lactic acid bacteria (LAB) employ sucrase-type enzymes to convert sucrose into homopolysaccharides consisting of either glucosyl units (glucans) or fructosyl units (fructans). The enzymes involved are labeled glucansucrases (GS) and fructansucrases (FS), respectively. The available molecular, biochemical, and structural information on sucrase genes and enzymes from various LAB and their fructan and α-glucan products is reviewed. The GS and FS enzymes are both glycoside hydrolase enzymes that act on the same substrate (sucrose) and catalyze (retaining) transglycosylation reactions that result in polysaccharide formation, but they possess completely different protein structures. GS enzymes (family GH70) are large multidomain proteins that occur exclusively in LAB. Their catalytic domain displays clear secondary-structure similarity with α-amylase enzymes (family GH13), with a predicted permuted (β/α)8 barrel structure for which detailed structural and mechanistic information is available. Emphasis now is on identification of residues and regions important for GS enzyme activity and product specificity (synthesis of α-glucans differing in glycosidic linkage type, degree and type of branching, glucan molecular mass, and solubility). FS enzymes (family GH68) occur in both gram-negative and gram-positive bacteria and synthesize β-fructan polymers with either β-(2→6) (inulin) or β-(2→1) (levan) glycosidic bonds. Recently, the first high-resolution three-dimensional structures have become available for FS (levansucrase) proteins, revealing a rare five-bladed β-propeller structure with a deep, negatively charged central pocket. Although these structures have provided detailed mechanistic insights, the structural features in FS enzymes dictating the synthesis of either β-(2→6) or β-(2→1) linkages, degree and type of branching, and fructan molecular mass remain to be identified

    Variability in concentrations of potentially toxic elements in urban parks from six European cities

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    Use of a harmonised sampling regime has allowed comparison of concentrations of copper, chromium, nickel, lead and zinc in six urban parks located in different European cities differing markedly in their climate and industrial history. Wide concentrations ranges were found for copper, lead and zinc at most sites, but for chromium and nickel a wide range was only seen in the Italian park, where levels were also considerably greater than in other soils. As might be expected, the soils from older cities with a legacy of heavy manufacturing industry (Glasgow, Torino) were richest in potentially toxic elements (PTEs); soils from Ljubljana, Sevilla and Uppsala had intermediate metal contents, and soils from the most recently established park, in the least industrialised city (Aveiro), displayed lowest concentrations. When principal component analysis was applied to the data, associations were revealed between pH and organic carbon content; and between all five PTEs. When pH and organic carbon content were excluded from the PCA, a distinction became clear between copper, lead and zinc (the "urban" metals) on the one hand, and chromium and nickel on the other. Similar results were obtained for the surface (0-10 cm depth) and sub-surface (10-20 cm depth) samples. Comparisons with target or limit concentrations were limited by the existence of different legislation in different countries and the fact that few guidelines deal specifically with public-access urban soils intended for recreational use

    Colloidal particles at a nematic-isotropic interface: effects of confinement

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    When captured by a flat nematic-isotropic interface, colloidal particles can be dragged by it. As a result spatially periodic structures may appear, with the period depending on a particle mass, size, and interface velocity~\cite{west.jl:2002}. If liquid crystal is sandwiched between two substrates, the interface takes a wedge-like shape, accommodating the interface-substrate contact angle and minimizing the director distortions on its nematic side. Correspondingly, particles move along complex trajectories: they are first captured by the interface and then `glide' towards its vertex point. Our experiments quantify this scenario, and numerical minimization of the Landau-de Gennes free energy allow for a qualitative description of the interfacial structure and the drag force.Comment: 7 pages, 9 figure

    Practical value of the semi-quantitative Rainsford-Davies test for the determination of phenol in the urine of workers exposed to benzene vapour

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    Ispitana je vrijednost semikvantitativnog fenol testa u odnosu na kvantitativne analize. Istraživanja su provedena na grupi od 50 radnika eksponiranih parama benzena jedne zagrebačke tvornice lijekova. Kontrolna grupa bilo je 20 radnika iste tvornice. Reakcija testa se temelji na stvaranju plave boje između prisutnog fenola i 2,6-diklorkinon-4-klorimida (Gibbsov reagens) u lužnatom mediju (pH 10,15). Intenzitet boje je linearno proporcionalan koncentraciji fenola u urinu, odnosno koncentraciji benzena u zraku. Rezultati su pokazali da semikvantitativno određivanje daje vrlo dobar uvid u grupnu i individualnu ekspoziciju parama benzena. Zbog jednostavnosti i brzine izvođenja, ta metoda ima praktičnu prednost pred drugima.The value of the semi-quantitative phenol test in relation to quantitative analysis was analysed in a group of 50 workers of a Zagreb pharmaceutical works exposed to benzene vapour. The control group consisted of 20 workers of the same factory. The reaction of the test is based on the production of a blue colour by phenol and 2,6-dichloroquinone-4-chloroimide (the Gibbs reagent) in an alkaline medium (pH 10.15). The colour intensity is in a linear proportion to the concentration of phenol in the urine and of benzene in the air, respectively. The results have shown that the semi-quantitative determination of phenol in relation to the quantitative one gives a very good insight into the colective and individual exposure to benzene vapour. The procedure is simple and quick, and for this reason of more practical value than other methods

    Diagnostic Interview for Genetic Studies: validity and reliability of the Croatian version

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    OBJECTIVE: To test the validity and reliability of the Diagnostic Interview for Genetic Studies (DIGS) in patients with mental illness in Croatia. MATERIALS AND METHODS: Following translation, back-translation, and pilot testing, the Croatian version of DIGS (CRO-DIGS) was administered to a total of 150 inpatients and outpatients diagnosed at the Clinical Hospital in Split with bipolar and major depressive disorder (n=56), schizophrenia and schizoaffective disorder (n=62), and alcohol dependence or use disorders (n=32). Initial testing was performed independently by one interviewer and one observer blinded to the diagnosis, and a retest was performed after 8 weeks by a third examiner. RESULTS: The validity of CRO-DIGS was high (κ=0.916), with an excellent inter-rater (κ=0.824) reliability, especially for bipolar disorder (κ=0.956). Following an 8 week test-retest interval, the reliability for all diagnoses was found to be excellent (κ=0.843). CONCLUSION: Our study has shown excellent validity and reliability of the Croatian version of DIGS, making it a promising instrument to assess mental illness of patients. The development of a valid and reliable diagnostic tool such as the CRO-DIGS will considerably advance the scientific communities' ability to carry out genetic studies of psychiatric illness in the region
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