Transglycosylation Activity of Engineered Bifidobacterium Lacto-N-Biosidase Mutants at Donor Subsites for Lacto-N-Tetraose Synthesis

The health benefits of human milk oligosaccharides (HMOs) make them attractive targets as supplements for infant formula milks. However, HMO synthesis is still challenging and only two HMOs have been marketed. Engineering glycoside hydrolases into transglycosylases may provide biocatalytic routes to the synthesis of complex oligosaccharides. Lacto-N-biosidase from Bifidobacterium bifidum (LnbB) is a GH20 enzyme present in the gut microbiota of breast-fed infants that hydrolyzes lacto-N-tetraose (LNT), the core structure of the most abundant type I HMOs. Here we report a mutational study in the donor subsites of the substrate binding cleft with the aim of reducing hydrolytic activity and conferring transglycosylation activity for the synthesis of LNT from p-nitrophenyl β-lacto-N-bioside and lactose. As compared with the wt enzyme with negligible transglycosylation activity, mutants with residual hydrolase activity within 0.05% to 1.6% of the wild-type enzyme result in transglycosylating enzymes with LNT yields in the range of 10–30%. Mutations of Trp394, located in subsite -1 next to the catalytic residues, have a large impact on the transglycosylation/hydrolysis ratio, with W394F being the best mutant as a biocatalyst producing LNT at 32% yield. It is the first reported transglycosylating LnbB enzyme variant, amenable to further engineering for practical enzymatic synthesis of LNT.     

The propagation of pressure in a gelled waxy oil pipeline as studied by particle imaging velocimetry

Paraffinic crude oils in pipelines may form waxy gels during flow shutdowns. These gels can be dislodged by applying pressure if the wall shear stress, proportional to the local pressure gradient, exceeds the gel yield stress. The simplest models assume that the axial pressure profile becomes linear immediately after a jump in upstream pressure, but this fails to account for gel time‐dependent rheology or the effect of gel voids on pressure wave propagation. To investigate the former factor, pressure profile and particle imaging velocimetry (PIV) measurements were performed on a model oil gelled under pressure to reduce void formation. After a jump in upstream pressure to a value insufficient to restart flow, the axial pressure profile becomes linear in a two‐step process, with an immediate small rise in downstream pressure followed by a time‐delayed jump. The local downstream gel deformation measured by PIV exhibits similar two‐step time dependence. © 2011 American Institute of Chemical Engineers AIChE J, 2012     

Characterization of stabilized porous magnetite core–shell nanogel composites based on crosslinked acrylamide/sodium acrylate copolymers

Stabilized and dispersed superparamagnetic porous nanogels based on sodium acrylate (AA‐Na) and acrylamide (AM) in a surfactant‐free aqueous system were synthesized via solution polymerization at room temperature. The formation of magnetite nanoparticles was confirmed and their properties characterized using Fourier transform infrared spectroscopy. Extensive characterization of the magnetic polymer particles using transmission electron microscopy (TEM), dynamic light scattering and zeta potential measurements revealed that Fe₃O₄ nanoparticles were incorporated into the shells of poly(AM/AA‐Na). The average particle size was 5–8 nm as determined from TEM. AM/AA‐Na nanoparticles with a diameter of about 11 nm were effectively assembled onto the negatively charged surface of the as‐synthesized Fe₃O₄ nanoparticles via electrostatic interaction. Crosslinked magnetite nanocomposites were prepared by in situ development of surface‐modified magnetite nanoparticles in an AM/AA‐Na hydrogel. Scanning electron microscopy was used to study the surface morphology of the prepared composites. The morphology, phase composition and crystallinity of the prepared nanocomposites were characterized. Atomic force microscopy and argon adsorption–desorption measurements of Fe₃O₄.AM/AA indicated that the architecture of the polymer network can be a hollow porous sphere or a solid phase, depending on the AA‐Na content. © 2013 Society of Chemical Industry     

Influence of very long aging on the relaxation behavior of flame‐retardant printed circuit board epoxy composites under mechatronic conditions

Very long aging times, up to 15,100 h (629 days) at 110°C, were achieved on flame‐retardant printed circuit board laminates commonly used in automotive design. This composite was fabricated from glass fibers embedded in an epoxy resin. Aging was performed in an oven under an air atmosphere at a temperature lower than the glass‐transition temperature. Temperature‐modulated differential scanning calorimetric analysis was used to investigate the influence of such aging on the glass‐transition phenomena. A new amorphous phase appeared during aging. By extending the analysis to samples collected at different thicknesses, we demonstrated the existence of a time‐dependent gradient of the properties. A skin–core structure was evidenced, and this slowed down oxidation and allowed physical aging to occur in the bulk sample. An exponential law described the variations of the glass‐transition of the new external compound. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 786‐792, 2013     

Tolerance modelling of vibrations of thin functionally graded plates

A problem of a modelling of vibrations of thin plates with a functionally graded macrostructure and a tolerance-periodic microstructure in planes parallel to the plate midplane is analysed. The proposed model, based on the Kirchhoff plate theory assumptions and additional hypothesis of the tolerance averaging technique [Wo?niak et al., editors. Thermomechanics of heterogeneous solids and structures. Tolerance averaging approach. ?ód?, University Press, Technical University of ?ód?; 2008], describes the effect of the microstructure size on dynamic behaviour of the plate. In this paper, as an example there are analysed free vibration frequencies of a functionally graded plate band. These frequencies are obtained in the framework of two proposed models—the tolerance model and the asymptotic model, using the known Ritz method.     

Transfer of thallium from rape seed to rape oil is negligible and oil is fit for human consumption

Rape and other Brassicaceae family plants can accumulate appreciable amounts of thallium from the soil. Because some species of this family are common crops utilised as food for direct consumption or raw materials for food production, thallium can enter the food chain. A useful method for thallium determination is inductively coupled plasma mass spectrometry. The limit of detection (0.2 pg ml^–1 Tl or 0.02 ng g^–1 Tl, taking in the account dilution during sample decomposition) found in the current study was very low, and the method can be used for ultra-trace analysis. Possible transfer of thallium from rape seed to the rape oil was investigated in two ways. The balance of thallium in rape seed meal (content 140–200 ng g^–1 Tl) and defatted rape seed meal indicated that thallium did not pass into the oil (p < 0.05). Moreover, the analyses of thallium in six kinds of edible rape seed oil and three kinds of margarines showed that the amount of thallium in rape seed oil is negligible.