38 research outputs found

    Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species

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    Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. We have previously characterized the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB. GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. The reaction kinetics demonstrated that GnbG can convert 69 ± 4 and 71 ± 1 % of o-nitrophenyl-β-D-galactopyranoside into LNB and GNB, respectively. Those reactions were performed in a semi-preparative scale, and the synthesized disaccharides were purified. The maximum yield obtained for LNB was 10.7 ± 0.2 g/l and for GNB was 10.8 ± 0.3 g/l. NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. The purified sugars were subsequently tested for their potential prebiotic properties using Lactobacillus species. The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants

    Screening potential pests of Nordic coniferous forests associated with trade in ornamental plants

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    Plant pests moved along with the trade in ornamental plants could pose a threat to forests. In this study plant pests potentially associated with this pathway were screened to identify pests that could pose a high risk to the coniferous forests of Finland, Sweden and Norway. Specifically, the aim was to find pests that potentially could fulfil the criteria to become regulated as quarantine pests. EPPO’s commodity study approach, which includes several screening steps, was used to identify the pests that are most likely to become significant pests of Picea abies or Pinus sylvestris. From an initial list of 1062 pests, 65 pests were identified and ranked using the FinnPRIO model, resulting in a top list of 14 pests, namely Chionaspis pinifoliae, Coleosporium asterum s.l., Cytospora kunzei, Dactylonectria macrodidyma, Gnathotrichus retusus, Heterobasidion irregulare, Lambdina fiscellaria, Orgyia leucostigma, Orthotomicus erosus, Pseudocoremia suavis, Tetropium gracilicorne, Toumeyella parvicornis, Truncatella hartigii and Xylosandrus germanus. The rankings of the pests, together with the collected information, can be used to prioritize pests and pathways for further assessment

    The ancestral role of ATP hydrolysis in type II topoisomerases: prevention of DNA double-strand breaks

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    Type II DNA topoisomerases (topos) catalyse changes in DNA topology by passing one double-stranded DNA segment through another. This reaction is essential to processes such as replication and transcription, but carries with it the inherent danger of permanent double-strand break (DSB) formation. All type II topos hydrolyse ATP during their reactions; however, only DNA gyrase is able to harness the free energy of hydrolysis to drive DNA supercoiling, an energetically unfavourable process. A long-standing puzzle has been to understand why the majority of type II enzymes consume ATP to support reactions that do not require a net energy input. While certain type II topos are known to ‘simplify’ distributions of DNA topoisomers below thermodynamic equilibrium levels, the energy required for this process is very low, suggesting that this behaviour is not the principal reason for ATP hydrolysis. Instead, we propose that the energy of ATP hydrolysis is needed to control the separation of protein–protein interfaces and prevent the accidental formation of potentially mutagenic or cytotoxic DSBs. This interpretation has parallels with the actions of a variety of molecular machines that catalyse the conformational rearrangement of biological macromolecules
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