Effect of Salts on Laccase-Catalyzed Polymerization of Lignosulfonate.

Enzymatic polymerization of lignosulfonate (LS) has a high potential for various applications ranging from coatings to adhesives. Here, the effect of different ions in low concentrations on enzymatic polymerization of LS was investigated, including salt solutions consisting of mono- and dicarboxylic acids, sulfate, phosphate and chloride with sodium as counter ion. LS polymerization was followed by viscometry and size exclusion (SEC) chromatography. Interestingly, there was only a small effect of ions on the activity of the laccase on standard substrate ABTS, while the effect on polymerization of LS was substantially different. The presence of acetate led to a 39 % higher degree of polymerization (DP) for LS. Small angle X-ray scattering (SAXS) revealed that the structure of the enzyme was largely unaffected by the ions, while the determination of the zeta potential showed that those ions conveying higher negative surface charges onto LS particles showed lower DPs, than those not affecting the surface charge. Further, electron paramagnetic resonance (EPR) spectroscopy showed 5-times higher intensity in phenoxyl radicals for the monovalent ions compared to the divalent ones. It was concluded that the DPs of LS could be tuned in the presence of certain ions, by facilitating the interaction between the laccase substrate-binding site and the LS molecules.

Ultrasonic fatigue of unfilled and carbon nanotube (CNT) reinforced polyetheretherketone (PEEK).

Fatigue properties of polyetheretherketone (PEEK) and multiwall carbon nanotube (CNT) reinforced PEEK were investigated with the ultrasonic fatigue testing method. Lifetimes were measured in the high and very high cycle fatigue regime at resonance frequency 19 kHz and load ratio R = -1. Pulse-pause loading served to avoid specimen self-heating and led to effective cycling frequencies in the range from several hundred Hz to about two kHz. Stress amplitude for 50 % fracture probability at 109 cycles is 21.2 ± 4.3 MPa for unreinforced PEEK (22 % of its tensile strength) and 33.5 ± 3.5 MPa for CNT reinforced PEEK (33 % of its tensile strength). Servohydraulic fatigue tests at 22 Hz with CNT reinforced PEEK delivered fatigue lifetimes comparable to ultrasonic tests, i.e. no frequency effect and no influence of load versus displacement control was observed. Keeping specimen temperature far below the glass transition temperature, ultrasonic fatigue testing of a high temperature resistant plastic was successfully implemented.

Dispersion State and Damage of Carbon Nanotubes and Carbon Nanofibers by Ultrasonic Dispersion: A Review.

Dispersion of carbon nanotubes and carbon nanofibers is a crucial processing step in the production of polymer-based nanocomposites and poses a great challenge due to the tendency of these nanofillers to agglomerate. Besides the well-established three-roll mill, the ultrasonic dispersion process is one of the most often used methods. It is fast, easy to implement, and obtains considerably good results. Nevertheless, damage to the nanofibers due to cavitation may lead to shortening and changes in the surface of the nanofillers. The proper application of the sonicator to limit damage and at the same time enable high dispersion quality needs dedicated knowledge of the damage mechanisms and characterization methods for monitoring nano-particles during and after sonication. This study gives an overview of these methods and indicates parameters to be considered in this respect. Sonication energy rather than sonication time is a key factor to control shortening. It seems likely that lower powers that are induced by a broader tip or plate sonicators at a longer running time would allow for proper dispersions, while minimizing damage.

Pore Development during the Carbonization Process of Lignin Microparticles Investigated by Small Angle X-ray Scattering.

Application of low-cost carbon black from lignin highly depends on the materials properties, which might by determined by raw material and processing conditions. Four different technical lignins were subjected to thermostabilization followed by stepwise heat treatment up to a temperature of 2000 °C in order to obtain micro-sized carbon particles. The development of the pore structure, graphitization and inner surfaces were investigated by X-ray scattering complemented by scanning electron microscopy and FTIR spectroscopy. Lignosulfonate-based carbons exhibit a complex pore structure with nanopores and mesopores that evolve by heat treatment. Organosolv, kraft and soda lignin-based samples exhibit distinct pores growing steadily with heat treatment temperature. All carbons exhibit increasing pore size of about 0.5-2 nm and increasing inner surface, with a strong increase between 1200 °C and 1600 °C. The chemistry and bonding nature shifts from basic organic material towards pure graphite. The crystallite size was found to increase with the increasing degree of graphitization. Heat treatment of just 1600 °C might be sufficient for many applications, allowing to reduce production energy while maintaining materials properties.

Metal-Insulator Transition of Ultrathin Sputtered Metals on Phenolic Resin Thin Films: Growth Morphology and Relations to Surface Free Energy and Reactivity.

Nanostructured metal assemblies on thin and ultrathin polymeric films enable state of the art technologies and have further potential in diverse fields. Rational design of the structure-function relationship is of critical importance but aggravated by the scarcity of systematic studies. Here, we studied the influence of the interplay between metal and polymer surface free energy and reactivity on the evolution of electric conductivity and the resulting morphologies. In situ resistance measurements during sputter deposition of Ag, Au, Cu and Ni films on ultrathin reticulated polymer films collectively reveal metal-insulator transitions characteristic for Volmer-Weber growth. The different onsets of percolation correlate with interfacial energy and energy of adhesion weakly but as expected from ordinary wetting theory. A more pronounced trend of lower percolation thickness for more reactive metals falls in line with reported correlations. Ex situ grazing incidence small angle X-ray scattering experiments were performed at various thicknesses to gain an insight into cluster and film morphology evolution. A novel approach to interpret the scattering data is used where simulated pair distance distributions of arbitrary shapes and arrangements can be fitted to experiments. Detailed approximations of cluster structures could be inferred and are discussed in view of the established parameters describing film growth behavior.

Solvent-Free Ultrasonic Dispersion of Nanofillers in Epoxy Matrix.

Dispersion of carbon nanotubes and carbon nanofibers is a crucial processing step in the production of polymer-based nanocomposites and poses a great challenge due to the tendency of nanofillers to agglomerate. One of the most effective methods for dispersion is the use of a three-roll mill, which is a well-established method and results in agglomerates below 5 µm. Nevertheless, this process is time-consuming and thus a limiting factor for industrial applications. Our aim was to establish an easy and efficient ultrasonic dispersion process, characterize the dispersion parameters, and compare both methods, ultrasonication and the three-roll mill. We applied rheological tests and analyzed the agglomerate sizes by an image fit of the microscopy images. All these analyses combined deliver a valuable set of information about the dispersion's quality and, therefore, allows the improvement and further adaptation of the dispersion process.

Insight into the nanostructure of anisotropic cellulose aerogels upon compression.

Cellulose II aerogels are a highly porous class of biobased ultra-light-weight materials. They consist of interlinked networks of loosely aggregated cellulose fibrils. The latter typically have random orientation due to spontaneous phase separation triggered by addition of antisolvent to moleculardisperse cellulose solutions. Deceleration of phase separation has been recently proposed as a novel approach towards aerogels featuring preferred cellulose orientation. Here, we investigate the mechanical response of such oriented cellulose aerogels towards load up to 80% compression. Stress-strain curves were recorded and in situ small angle X-ray scattering (SAXS) was performed during compression test to obtain information about the structural alterations of the aerogel fibril networks on the nano-scale related to deformation. Using SAXS in addition, structural changes can be followed in much more detail than by recording stress-strain curves alone. Buckling and coalescence of fibers and a change in fibril orientation can be related to certain regimes in the stress-strain curve. If the loading axis is oriented parallel to the network orientation the aerogels show higher resilience towards the compression.

High-resolution large-area imaging of nanoscale structure and mineralization of a sclerosing osteosarcoma in human bone.

Osteosarcoma is the most common primary bone cancer type in humans. It is predominantly found in young individuals, with a second peak later in life. The tumour is formed by malignant osteoblasts and consists of collagenous, sometimes also mineralized, bone matrix. While the morphology of osteosarcoma has been well studied, there is virtually no information about the nanostructure of the tumour and changes in mineralization on the nanoscale level. In the present paper, human bone tissue inside, next to and remote from a sclerosing osteosarcoma was studied with small angle x-ray scattering, x-ray diffraction and electron microscopy. Quantitative evaluation of nanostructure parameters was combined with high resolution, large area mapping to obtain microscopic images with nanostructure parameter contrast. It was found that the tumour regions were characterized by a notable reduction in mineral particle size, while the mineral content was even higher than that in normal bone. Furthermore, the normal preferential orientation of mineral particles along the longitudinal direction of corticalis or trabeculae was largely suppressed. Also the bone mineral crystal structure was affected: severe crystal lattice distortions were detected in mineralized tumour tissue pointing to a different ion substitution of hydroxyl apatite in tumorous tissue than in healthy tissue.

Impact of storage on the physico-chemical properties of microparticles comprising a hydrogenated vegetable oil matrix and different essential oil concentrations.

Microparticles made from hydrogenated sunflower oil without essential oil and with different essential oil concentrations (75-300 g/kg; g of essential oil per kg of microparticles) were stored for 1 or 2 months at 25 or 37 °C. Before and after storage the essential oil concentration, flowability, optical appearance, melting behaviour and crystalline structure of the microparticles were investigated. Essential oil recovery, melting behaviour and crystalline structure were identical for the essential oil containing microparticles and were not affected during storage. The surface structure of the microparticles varied with their essential oil concentration. While the particles containing 75 g/kg essential oil were covered by erect fat crystals, those with 225 g/kg and higher were mostly smooth with some round shaped dumps. However, the surface of all essential oil containing microparticle batches had reached their final stage after production already and did not change during storage. Microparticles without essential oil presented two melting peaks; all microparticle batches with essential oil had one peak. Peaks in the X-ray scattering powder diffraction signal of the essential oil-free microparticles after production can be associated with the α-form of the hydrogenated vegetable oil. During storage, a conversion of the α-form to the stable ß-form was observed. Microscopy showed that these microparticles also developed strong fat crystals throughout storage. The triglycerides in microparticles with essential oil seem to directly take on the stable ß-form. The formation of robust microparticle agglomerates during storage was prevalently observed for the fat crystal forming product batches, meaning the products without or with low essential oil concentration.

Enhanced Cu and Cd sorption after soil aging of woodchip-derived biochar: What were the driving factors?

Biochar (BC) is increasingly tested as a soil amendment for immobilization of heavy metals (HMs) and other pollutants. In our study, an acidic soil amended with wood chip-derived BC showed strongly enhanced Cu and Cd sorption after 15 months of aging under greenhouse conditions. X-ray absorption near edge structure suggested formation of Cu(OH)2 and CuCO3 and upon aging increasingly Cu sorption to the BC organic phase (from 9.2% to 40.7%) as main binding mechanisms of Cu on the BCs. In contrast, Cd was predominantly bound as CdCO3 on the BCs even after 15 months (82.7%). We found indications by mid-infrared spectroscopy that the formation of organic functional groups plays a role for increased HM sorption on aged BCs. Yet, our data suggest that the accessibility of BC's pore network and reactive surfaces is likely to be the overriding factor responsible for aging-related changes in HM sorption capacity, rather than direct interactions of HMs with oxidized functional groups. We observed highly weathered BC surface structures with scanning electron microscopy along with strongly increased wettability of the BCs after 15 months of soil aging as indicated by a decrease of water contact angles (from 62.4° to 4.2°).

Electrically-Conductive Sub-Micron Carbon Particles from Lignin: Elucidation of Nanostructure and Use as Filler in Cellulose Nanopapers.

Carbon particles were produced from kraft lignin through carbonization of perfectly spherical, sub-micron beads obtained by aerosol flow. The structure of the resulting carbon particles was elucidated and compared to that derived from commercially available technical lignin powder, which is undefined in geometry. In addition to the smaller diameters of the lignin beads (<1 µm) compared to those of the lignin powder (100 µm), the former displayed a slightly higher structural order as revealed by X-ray diffraction and Raman spectroscopy. With regard to potential application in composite structures, the sub-micron carbon beads were clearly advantageous as a filler of cellulose nanopapers, which displayed better mechanical performance but with limited electrical conductivity. Compression sensing was achieved for this nanocomposite system.

Acid Free Oxidation and Simple Dispersion Method of MWCNT for High-Performance CFRP.

Carbon nanotubes (CNT) provide an outstanding property spectrum which can be used to improve a wide range of materials. However, the transfer of properties from the nanoscale to a macroscopic material is a limiting factor. Different approaches of functionalizing the surface of a CNT can improve the interaction with the surrounding matrix but is connected to difficult and expensive treatments, which are usually inconvenient for industrial applications. Here, a simple and eco-friendly method is presented for the oxidation of CNT, where hydrogen peroxide (H2O2) is the only chemical needed and no toxic emissions are released. Also, the extensive step of the incorporation of CNT to an epoxy matrix is simplified to an ultrasonic dispersion in the liquid hardener component. The effectiveness is proven by mechanical tests of produced CNT/CFRP and compared to a conventional processing route. The combination of those simple and cost efficient strategies can be utilized to produce multiscale composites with improved mechanical performance in an ecological and economical way.

Antibody adsorption in protein-A affinity chromatography - in situ measurement of nanoscale structure by small-angle X-ray scattering.

Protein-A chromatography is the most widely used chromatography step in downstream processing of antibodies. A deeper understanding of the influence of the surface topology on a molecular/nanoscale level on adsorption is essential for further improvement. It is not clear if the binding is homogenous throughout the entire bead network. We followed the protein absorption process and observed the formation of a protein layer on fibers of chromatography resin in a time-resolved manner in nanoscale. To characterize the changes in the antibody-protein-A ligand complex, small angle X-ray scattering was employed using a miniaturized X-ray-transparent chromatography column packed with a MabSelect SuRe resin. Antibody-free MabSelect SuRe resin fiber had an average radius of 12 nm and the protein layer thickness resulting from antibody adsorption was 5.5 and 10.4 nm for fiber and junctions, respectively under applied native conditions. We hypothesize that an average of 1.2 antibodies were adsorbed per protein-A ligand tetramer bound to the outermost units. In contrast to previous studies, it was therefore possible for the first time to directly correlate the nanostructure changes inside the column, which is otherwise a black box, with the adsorption and elution process.

Self-Assembly of Cellulose in Super-Cooled Ionic Liquid under the Impact of Decelerated Antisolvent Infusion: An Approach toward Anisotropic Gels and Aerogels.

Assembly of (bio)polymers into long-range anisotropic nanostructured gels and aerogels is of great interest in advanced material engineering since it enables directional tuning of properties, such as diffusivity, light, heat, and sound propagation, cell proliferation, and mechanical properties. Here we present an approach toward anisotropic cellulose II gels and aerogels that employs specific diffusion and phase separation phenomena occurring during decelerated infusion of an antisolvent into isotropic supercooled solutions of cellulose in an ionic liquid to effectuate supramolecular assembly of cellulose in anisotropic colloidal network structures. At the example of the distillable ionic liquid 1,1,3,3-tetramethylguanidinium acetate, the antisolvent ethanol, and spherocylindrical porous molds, we demonstrate that the proposed facile, environmental-benign and versatile route affords gels and aerogels whose specific anisotropic nanomorphology and properties reflect the preferred supramolecular cellulose orientation during phase separation, which is perpendicular to the direction of antisolvent diffusion. Comprehensive X-ray scattering experiments revealed that the (aero)gels are composed of an interconnected, fibrous, highly crystalline (CrI ≈ 72%), cellulose II with a cross-sectional Guinier radius of the struts of about 2.5 nm, and an order parameter gradient from about 0.1 to 0.2. The obtained gels and aerogels feature high specific surface areas (350-630 m2 g-1) and excellent mechanical properties like high toughness (up to 471 kJ m-3 for a 60% compression, ρB = 80 mg cm-3) and resilience (up to 13.4 kJ m-3, ρB = 65 mg cm-3).

Formation of essential oil containing microparticles comprising a hydrogenated vegetable oil matrix and characterisation thereof.

Microparticles with different essential oil concentrations 0, 75, 150, 225 and 300 gkg-1, (g of essential oil per kg of microparticles), were produced by dispersing the essential oils within a hydrogenated vegetable fat matrix and forming spherical solid particles by spray-chilling. Size distribution, flowability, surface structure, essential oil recovery, melting properties and crystallinity of the microparticles were determined. With over 225 gkg-1 essential oil the microparticle surface became stickier, their flowability was reduced and the size distribution broadened. Gas chromatography showed that the essential oil recovery was always above 85% v/v. The surface structure of the microparticles was strongly affected by the essential oil concentration being smooth (225 gkg-1), comprising round-shaped dumps (300 gkg-1) or showing fat blooming (0, 75, 150 gkg-1). With essential oil, the formation of the ß-polymorphic form of the triglycerides was supported leading to changes in the melting behaviour and the crystalline structure.

Photon Energy Becomes the Third Dimension in Crystallographic Texture Analysis.

Conventional analysis of the preferred orientation of crystallites (crystallographic texture) involves X-ray diffraction with area detectors and 2D data output. True 3D, spatially resolved information requires sample rotation in the beam, thus changing the probed volume, which introduces signal smearing and precludes the scanning of complex structures. This obstacle has been overcome by energy-dispersive Laue diffraction. A method has been devised to reach a large portion of reciprocal space and translate the X-ray photon energy into the missing third dimension of space. Carbon fibers and lobster exoskeleton as examples of biomineralized tissue have been analyzed. The major potential of this method lies in its "one-shot" nature and the direct 3D information requiring no previous knowledge of the sample. It allows the texture of large samples with complex substructures to be scanned and opens up the conceptual possibility of following texture changes in situ, for example, during crystallization.

Impact of selected solvent systems on the pore and solid structure of cellulose aerogels.

The impact of selected cellulose solvent systems based on the principal constituents tetrabutylammonium fluoride (TBAF), 1-ethyl-3-methyl-1H-imidazolium-acetate, N-methylmorpholine-N-oxide, or calcium thiocyanate octahydrate (CTO) on the properties of cellulose II aerogels prepared from these solvent systems has been investigated as a means towards tailoring cellulose aerogel properties with respect to specific applications. Cotton linters were used as representative plant cellulose. Cellulose was coagulated from solutions with comparable cellulose content, and dried with supercritical carbon dioxide after solvent exchange. The resulting bulk aerogels were comprehensively morphologically and mechanically tested to relate structure and mechanical properties. Different solvent systems caused considerable differences in the properties of the bulk samples, such as internal surface area (nitrogen sorption), morphology, porosity (He pycnometry, thermoporosimetry), and mechanical stability (compression testing). The results of SAXS, WAXS, and solid-state 13C NMR spectroscopy suggest that this is due to different mechanisms of cellulose self-assembling on the supramolecular and nanostructural level, respectively, as reflected by the broad ranges of cellulose crystallinity, fibril diameter, fractal dimension and skeletal density. Both solid state NMR and WAXS experiments confirmed the sole existence of the cellulose II allomorph for all aerogels, with crystallinity reaching a maximum of 46-50 % for CTO-derived aerogels. Generally, higher fibril diameter, degree of crystallinity, hence increased skeletal density were associated with good preservation of shape and dimension throughout conversion of lyogels to aerogels, and enhanced mechanical stability, but somewhat reduced specific surface area. Amorphous, yet highly rigid aerogels derived from TBAF/DMSO mixtures deviated from this trend, most likely due to their particular homogeneous and nanostructured morphology.

Carbon Microparticles from Organosolv Lignin as Filler for Conducting Poly(Lactic Acid).

Carbon microparticles were produced from organosolv lignin at 2000 °C under argon atmosphere following oxidative thermostabilisation at 250 °C. Scanning electron microscopy, X-ray diffraction, small-angle X-ray scattering, and electro-conductivity measurements revealed that the obtained particles were electrically conductive and were composed of large graphitic domains. Poly(lactic acid) filled with various amounts of lignin-derived microparticles showed higher tensile stiffness increasing with particle load, whereas strength and extensibility decreased. Electric conductivity was measured at filler loads equal to and greater than 25% w/w.

Core-Shell Structure of Monodisperse Poly(ethylene glycol)-Grafted Iron Oxide Nanoparticles Studied by Small-Angle X-ray Scattering.

The promising applications of core-shell nanoparticles in the biological and medical field have been well investigated in recent years. One remaining challenge is the characterization of the structure of the hydrated polymer shell. Here we use small-angle X-ray scattering (SAXS) to investigate iron oxide core-poly(ethylene glycol) brush shell nanoparticles with extremely high polymer grafting density. It is shown that the shell density profile can be described by a scaling model that takes into account the locally very high grafting density near the core. A good fit to a constant density region followed by a star-polymer-like, monotonously decaying density profile is shown, which could help explain the unique colloidal properties of such densely grafted core-shell nanoparticles. SAXS experiments probing the thermally induced dehydration of the shell and the response to dilution confirmed that the observed features are associated with the brush and not attributed to structure factors from particle aggregates. We thereby demonstrate that the structure of monodisperse core-shell nanoparticles with dense solvated shells can be well studied with SAXS and that different density models can be distinguished from each other.

Enhanced cutinase-catalyzed hydrolysis of polyethylene terephthalate by covalent fusion to hydrophobins.

Cutinases have shown potential for hydrolysis of the recalcitrant synthetic polymer polyethylene terephthalate (PET). We have shown previously that the rate of this hydrolysis can be enhanced by the addition of hydrophobins, small fungal proteins that can alter the physicochemical properties of surfaces. Here we have investigated whether the PET-hydrolyzing activity of a bacterial cutinase from Thermobifida cellulosilytica (Thc_Cut1) would be further enhanced by fusion to one of three Trichoderma hydrophobins, i.e., the class II hydrophobins HFB4 and HFB7 and the pseudo-class I hydrophobin HFB9b. The fusion enzymes exhibited decreased kcat values on soluble substrates (p-nitrophenyl acetate and p-nitrophenyl butyrate) and strongly decreased the hydrophilicity of glass but caused only small changes in the hydrophobicity of PET. When the enzyme was fused to HFB4 or HFB7, the hydrolysis of PET was enhanced >16-fold over the level with the free enzyme, while a mixture of the enzyme and the hydrophobins led only to a 4-fold increase at most. Fusion with the non-class II hydrophobin HFB9b did not increase the rate of hydrolysis over that of the enzyme-hydrophobin mixture, but HFB9b performed best when PET was preincubated with the hydrophobins before enzyme treatment. The pattern of hydrolysis by the fusion enzymes differed from that of Thc_Cut1 as the concentration of the product mono(2-hydroxyethyl) terephthalate relative to that of the main product, terephthalic acid, increased. Small-angle X-ray scattering (SAXS) analysis revealed an increased scattering contrast of the fusion proteins over that of the free proteins, suggesting a change in conformation or enhanced protein aggregation. Our data show that the level of hydrolysis of PET by cutinase can be significantly increased by fusion to hydrophobins. The data further suggest that this likely involves binding of the hydrophobins to the cutinase and changes in the conformation of its active center.