How Resilient is Wood Xylan to Enzymatic Degradation in a Matrix with Kraft Lignin?

Despite the potential of lignocellulose in manufacturing value-added chemicals and biofuels, its efficient biotechnological conversion by enzymatic hydrolysis still poses major challenges. The complex interplay between xylan, cellulose, and lignin in fibrous materials makes it difficult to assess underlying physico- and biochemical mechanisms. Here, we reduce the complexity of the system by creating matrices of cellulose, xylan, and lignin, which consists of a cellulose base layer and xylan/lignin domains. We follow enzymatic degradation using an endoxylanase by high-speed atomic force microscopy and surface plasmon resonance spectroscopy to obtain morphological and kinetic data. Fastest reaction kinetics were observed at low lignin contents, which were related to the different swelling capacities of xylan. We demonstrate that the complex processes taking place at the interfaces of lignin and xylan in the presence of enzymes can be monitored in real time, providing a future platform for observing phenomena relevant to fiber-based systems.

Deciphering heterogeneous enzymatic surface reactions on xylan using surface plasmon resonance spectroscopy.

Xylans' unique properties make it attractive for a variety of industries, including paper, food, and biochemical production. While for some applications the preservation of its natural structure is crucial, for others the degradation into monosaccharides is essential. For the complete breakdown, the use of several enzymes is required, due to its structural complexity. In fact, the specificity of enzymatically-catalyzed reactions is guided by the surface, limiting or regulating accessibility and serving structurally encoded input guiding the actions of the enzymes. Here, we investigate enzymes at surfaces rich in xylan using surface plasmon resonance spectroscopy. The influence of diffusion and changes in substrate morphology is studied via enzyme surface kinetics simulations, yielding reaction rates and constants. We propose kinetic models, which can be applied to the degradation of multilayer biopolymer films. The most advanced model was verified by its successful application to the degradation of a thin film of polyhydroxybutyrate treated with a polyhydroxybutyrate-depolymerase. The herein derived models can be employed to quantify the degradation kinetics of various enzymes on biopolymers in heterogeneous environments, often prevalent in industrial processes. The identification of key factors influencing reaction rates such as inhibition will contribute to the quantification of intricate dynamics in complex systems.

Cellulose modified to host functionalities via facile cation exchange approach.

Properties of cellulose are typically functionalized by organic chemistry means. We progress an alternative facile way to functionalize cellulose by functional group counter-cation exchange. While ion-exchange is established for cellulose, it is far from exploited and understood beyond the most common cation, sodium. We build on our work that established the cation exchange for go-to alkali metal cations. We expand and further demonstrate the introduction of functional cations, namely, lanthanides. We show that cellulose nanocrystals (CNCs) carrying sulfate-half ester groups can acquire properties through the counter-cation exchange. Trivalent lanthanide cations europium (Eu3+), dysprosium (Dy3+) and gadolinium (Gd3+) were employed. The respective ions showed distinct differences in their ability of being coordinated by the sulfate groups; with Eu3+ fully saturating the sulfate groups while for Gd3+ and Dy3+, values of 82 and 41 % were determined by compositional analysis. CNCs functionalized with Eu3+ displayed red emission, those containing Dy3+ exhibited no optical functionality, while those with Gd3+ revealed significantly altered magnetic relaxation times. Using cation exchange to alter cellulose properties in various ways is a tremendous opportunity for modification of the abundant cellulose raw materials for a renewable future.

The European Polysaccharide Network of Excellence (EPNOE) research roadmap 2040: Advanced strategies for exploiting the vast potential of polysaccharides as renewable bioresources.

Polysaccharides are among the most abundant bioresources on earth and consequently need to play a pivotal role when addressing existential scientific challenges like climate change and the shift from fossil-based to sustainable biobased materials. The Research Roadmap 2040 of the European Polysaccharide Network of Excellence (EPNOE) provides an expert's view on how future research and development strategies need to evolve to fully exploit the vast potential of polysaccharides as renewable bioresources. It is addressed to academic researchers, companies, as well as policymakers and covers five strategic areas that are of great importance in the context of polysaccharide related research: (I) Materials & Engineering, (II) Food & Nutrition, (III) Biomedical Applications, (IV) Chemistry, Biology & Physics, and (V) Skills & Education. Each section summarizes the state of research, identifies challenges that are currently faced, project achievements and developments that are expected in the upcoming 20 years, and finally provides outlines on how future research activities need to evolve.

Anion-Specific Adsorption of Carboxymethyl Cellulose on Cellulose.

Integration of fiber modification step with a modern pulp mill is a resource efficient way to produce functional fibers. Motivated by the need to integrate polymer adsorption with the current pulping system, anion-specific effects in carboxymethylcellulose (CMC) adsorption have been studied. The QCM-D adsorption experiments revealed that CMC adsorption to the cellulose model surface is prone to anion-specific effects. A correlation was observed between the adsorbed CMC and the degree of hydration of the co-ions present in the magnesium salts. The presence of a chaotropic co-ion such as nitrate increased the adsorption of CMC on cellulose compared to the presence of the kosmotropic sulfate co-ion. However, anion-specificity was not significant in the case of salts containing zinc cations. The hydration of anions determines the distribution of the ions at the interface. Chaotropic ions, such as nitrates, are likely to be distributed near the chaotropic cellulose surface, causing changes in the ordering of water molecules and resulting in greater entropy gain once released from the surface, thus increasing CMC adsorption.

Morphology and swelling of thin films of dialcohol xylan.

Polysaccharides are excellent network formers and are often processed into films from water solutions. Despite being hydrophilic polysaccharides, the typical xylans liberated from wood are sparsely soluble in water. We have previously suggested that an additional piece to the solubilization puzzle is modification of the xylan backbone via oxidative cleavage of the saccharide ring. Here, we demonstrate the influence of the degree of modification, i.e., degree of oxidation (DO) on xylan solubilization and consequent film formation and stability. Oxidized and reduced wood xylans (i.e., dialcohol xylans) with the highest DO (77 %) within the series exhibited the smallest hydrodynamic diameter (dh) of 60 nm in dimethylsulfoxide (DMSO). We transferred the modified xylans into films credit to their established solubility and then quantified the film water interactions. Dialcohol xylans with intermediate DOs (42 and 63 %) did not form continuous films. The films swelled slightly when subjected to humidity. However, the film with the highest DO demonstrated a significant moisture uptake that depended on the film mass and was not observed with the other modified grades or with unmodified xylan.

Potential of Wood Hemicelluloses and Their Derivates as Food Ingredients.

A holistic utilization of all lignocellulosic wood biomass, instead of the current approach of using only the cellulose fraction, is crucial for the efficient, ecological, and economical use of the forest resources. Use of wood constituents in the food and feed sector is a potential way of promoting the global economy. However, industrially established food products utilizing such components are still scarce, with the exception of cellulose derivatives. Hemicelluloses that include xylans and mannans are major constituents of wood. The wood hemicelluloses are structurally similar to hemicelluloses from crops, which are included in our diet, for example, as a part of dietary fibers. Hence, structurally similar wood hemicelluloses have the potential for similar uses. We review the current status and future potential of wood hemicelluloses as food ingredients. We include an inventory of the extraction routes of wood hemicelluloses, their physicochemical properties, and some of their gastrointestinal characteristics, and we also consider the regulatory route that research findings need to follow to be approved for food solutions, as well as the current status of the wood hemicellulose applications on that route.

Spatioselective surface chemistry for the production of functional and chemically anisotropic nanocellulose colloids.

Maximizing the benefits of nanomaterials from biomass requires unique considerations associated with their native chemical and physical structure. Both cellulose nanofibrils and nanocrystals are extracted from cellulose fibers via a top-down approach and have significantly advanced materials chemistry and set new benchmarks in the last decade. One major challenge has been to prepare defined and selectively modified nanocelluloses, which would, e.g., allow optimal particle interactions and thereby further improve the properties of processed materials. At the molecular and crystallite level, the surface of nanocelluloses offers an alternating chemical structure and functional groups of different reactivity, enabling straightforward avenues towards chemically anisotropic and molecularly patterned nanoparticles via spatioselective chemical modification. In this review, we will explain the influence and role of the multiscale hierarchy of cellulose fibers in chemical modifications, and critically discuss recent advances in selective surface chemistry of nanocelluloses. Finally, we will demonstrate the potential of those chemically anisotropic nanocelluloses in materials science and discuss challenges and opportunities in this field.

Dynamic and Static Assembly of Sulfated Cellulose Nanocrystals with Alkali Metal Counter Cations.

Sulfate groups on cellulose particles such as cellulose nanocrystals (CNCs) provide colloidal stability credit to electrostatic repulsion between the like-charged particles. The introduction of sodium counter cations on the sulfate groups enables drying of the CNC suspensions without irreversible aggregation. Less is known about the effect of other counter cations than sodium on extending the properties of the CNC particles. Here, we introduce the alkali metal counter cations, Li+, Na+, K+, Rb+, and Cs+, on sulfated CNCs without an ion exchange resin, which, so far, has been a common practice. We demonstrate that the facile ion exchange is an efficient method to exchange to any alkali metal cation of sulfate half esters, with exchange rates between 76 and 89%. The ability to form liquid crystalline order in rest was observed by the presence of birefringence patterns and followed the Hofmeister series prediction of a decreasing ability to form anisotropy with an increasing element number. However, we observed the K-CNC rheology and birefringence as a stand-out case within the series of alkali metal modifications, with dynamic moduli and loss tangent indicating a network disruptive effect compared to the other counter cations, whereas observation of the development of birefringence patterns in flow showed the absence of self- or dynamically-assembled liquid crystalline order.

Xylan-cellulose thin film platform for assessing xylanase activity.

Enzymatic degradation of plant polysaccharide networks is a complex process that involves disrupting an intimate assembly of cellulose and hemicelluloses in fibrous matrices. To mimic this assembly and to elucidate the efficiency of enzymatic degradation in a rapid way, models with physicochemical equivalence to natural systems are needed. Here, we employ xylan-coated cellulose thin films to monitor the hydrolyzing activity of an endo-1,4-ß-xylanase. In situ surface plasmon resonance spectroscopy (SPRS) revealed a decrease in xylan areal mass ranging from 0.01 ± 0.02 to 0.52 ± 0.04 mg·m-2. The extent of digestion correlates to increasing xylanase concentration. In addition, ex situ determination of released monosaccharides revealed that incubation time was also a significant factor in degradation (P > 0.01). For both experiments, atomic force microscopy confirmed the removal of xylans from the cellulose thin films. We provide a new model platform that offers nanoscale sensitivity for investigating biopolymer interactions and their susceptibility to enzymatic hydrolysis.

Modification of xylan via an oxidation-reduction reaction.

Xylan is a biopolymer readily available from forest resources. Various modification methods, including oxidation with sodium periodate, have been shown to facilitate the engineering applications of xylan. However, modification procedures are often optimized for semicrystalline high molecular weight polysaccharide cellulose rather than for lower molecular weight and amorphous polysaccharide xylan. This paper elucidates the procedure for the periodate oxidation of xylan into dialdehyde xylan and its further reduction into a dialcohol form and is focused on the modification work up. The oxidation-reduction reaction decreased the molecular weight of xylan while increased the dispersity more than 50%. Unlike the unmodified xylan, all the modified grades could be solubilized in water, which we see essential for facilitating the future engineering applications of xylan. The selection of quenching and purification procedures and pH-adjustment of the reduction step had no significant effect on the degree of oxidation, molecular weight and only a minor effect on the hydrodynamic radius in water. Hence, it is possible to choose the simplest oxidation-reduction route without time consuming purification steps within the sequence.

Calcium Ion-Induced Structural Changes in Carboxymethylcellulose Solutions and Their Effects on Adsorption on Cellulose Surfaces.

The adsorption of carboxymethylcellulose (CMC) on cellulose surfaces is one of the most studied examples of the adsorption of an anionic polyelectrolyte on a like-charged surface. It has been suggested that divalent ions can act as a bridge between CMC chains and the surface of cellulose and enhance the CMC adsorption: they can, however, also alter the structure of CMCs in the solution. In previous investigations, the influence of cations on solution properties has been largely overlooked. This study investigates the effect of Ca2+ ions on the properties of CMC solutions as well as the influence on cellulose nanofibers (CNFs), which was studied by dynamic light scattering and correlated with the adsorption of CMC on a cellulose surface probed using QCM-D. The presence of Ca2+ facilitated the multichain association of CMC chains and increased the hydrodynamic diameter. This suggests that the adsorption of CMCs at high concentrations of CaCl2 is governed mainly by changes in solution properties rather than by changes in the cellulose surface. Furthermore, an entropy-driven mechanism has been suggested for the adsorption of CMC on cellulose. By comparing the adsorption of CMC from H2O and D2O, it was found that the release of water from the cellulose surface is driving the adsorption of CMC.

Fundamental aspects of the non-covalent modification of cellulose via polymer adsorption.

The increasing need for new material applications based on cellulose demands increased functional diversity and thus new functionalisation/modification approaches. The non-covalent modification of cellulose fibres via the adsorption of functional polymers has emerged as a promising route for tailoring the properties of material. This review focuses on fundamental aspects of polymer adsorption on cellulose surfaces, where the adsorption of polyelectrolytes and non-polyelectrolytes are treated separately. Adsorption studies on model surfaces as well as cellulose macro-fibres are reviewed. A correlation of the adsorption findings with the Scheutjens-Fleer polymer adsorption theory is provided, allowing the fundamentals behind the polymer adsorption phenomenon and its context in utilization of cellulose fibres to be understood.

Fat tissue equivalent phantoms for microwave applications by reinforcing gelatin with nanocellulose.

Tissue mimicking phantom materials with thermal and dielectric equivalence are vital for the development of microwave diagnostics and treatment. The current phantoms representing fat tissue are challenged by mechanical integrity at relevant temperatures coupled with complex production protocols. We have employed two types of nanocellulose (cellulose nanocrystals and oxidized cellulose nanocrystals) as reinforcement in gelatin stabilized emulsions for mimicking fat tissue. The nanocellulose-gelatin stabilized emulsions were evaluated for their dielectric properties, the moduli-temperature dependence using small deformation rheology, stress-strain behavior using large deformation, and their compliance to quality assurance guidelines for superficial hyperthermia. All emulsions had low permittivity and conductivity within the lower microwave frequency band, accompanied by fat equivalent thermal properties. Small deformation rheology showed reduced temperature dependence of the moduli upon addition of nanocellulose, independent of type. The cellulose nanocrystals gelatin reinforced emulsion complied with the quality assurance guidelines. Hence, we demonstrate that the addition of cellulose nanocrystals to gelatin stabilized emulsions has the potential to be used as fat phantoms for the development of microwave diagnostics and treatment.

Chemical Engineering Laboratory Projects in Student Teams in Real Life and Transformed Online: Viscose Fiber Spinning and Characterization.

Chemical engineering education comprises a complexity of technical skills that include learning processes that are currently relevant in industry. Despite being a rather old industrial process, the manufacturing of viscose fibers still accounts for the major fraction of all human-made cellulosic fibers worldwide. Here we describe a laboratory setup to introduce chemistry and engineering students into the principles of cellulose fiber spinning according to the viscose process. The setup for fiber spinning is kept simplistic and allows the experiments to be performed without professional spinning equipment. However, all of the steps are performed analogously to the industrial process. The professional setting in process and chemical engineering involves work on projects and in teams. Hence, we have incorporated the fiber spinning laboratory experiment in the context of working in teams on projects. We will also present our experience on transferring a real-life laboratory experiment online, as this is required at times that online education is preferred over real-life teaching.

Cellulose Nanocrystal Liquid Crystal Phases: Progress and Challenges in Characterization Using Rheology Coupled to Optics, Scattering, and Spectroscopy.

Cellulose nanocrystals (CNCs) self-assemble and can be flow-assembled to liquid crystalline orders in a water suspension. The orders range from nano- to macroscale with the contributions of individual crystals, their micron clusters, and macroscopic assemblies. The resulting hierarchies are optically active materials that exhibit iridescence, reflectance, and light transmission. Although these assemblies have the potential for future renewable materials, details about structures on different hierarchical levels that span from the nano- to the macroscale are still not unraveled. Rheological characterization is essential for investigating flow properties; however, bulk material properties make it difficult to capture the various length-scales during assembly of the suspensions, for example, in simple shear flow. Rheometry is combined with other characterization methods to allow direct analysis of the structure development in the individual hierarchical levels. While optical techniques, scattering, and spectroscopy are often used to complement rheological observations, coupling them in situ to allow simultaneous observation is paramount to fully understand the details of CNC assembly from liquid to solid. This Review provides an overview of achievements in the coupled analytics, as well as our current opinion about opportunities to unravel the structural distinctiveness of cellulose nanomaterials.

Oxidized xylan additive for nanocellulose films - A swelling modifier.

Polymeric wood hemicelluloses are depicted to join cellulose, starch and chitosan as key polysaccharides for sustainable materials engineering. However, the approaches to incorporate hemicelluloses in emerging bio-based products are challenged by lack of specific benefit, other than the biomass-origin, although their utilization would contribute to sustainable material use since they currently are a side stream that is not valorized. Here we demonstrate wood-xylans as swelling modifiers for neutral and charged nanocellulose films that have already entered the sustainable packaging applications, however, suffer from humidity sensitivity. The oxidative modification is used to modulate the water-solubility of xylan and hence enable adsorption in an aqueous environment. A high molecular weight grade, hence less water-soluble, adsorbed preferentially on the neutral surface while the adsorbed amount on a negatively charged surface was independent of the molecular weight, and hence, solubility. The adsorption of the oxidized xylans on a neutral cellulose surface resulted in an increase in the amount of water in the film while on the negatively charged cellulose the total amount of water decreased. The finding of synergy of two hygroscopic materials to decrease swelling in hydrophilic bio-polymer films demonstrates the oxidized macromolecule xylan as structurally functional component in emerging cellulose products.

How cellulose nanofibrils and cellulose microparticles impact paper strength-A visualization approach.

Cellulosic nanomaterials are in the focus of academia and industry to realize light-weight biobased materials with remarkable strength. While the effect is well known, the distribution of these nanomaterials are less explored, particularly for paper sheets. Here, we explore the 3D distribution of micro and nanosized cellulosic particles in paper sheets and correlate their extent of fibrillation to the distribution inside the sheets and subsequently to paper properties. To overcome challenges with contrast between the particles and the matrix, we attached probes on the cellulose nano/microparticles, either by covalent attachment of fluorescent dyes or by physical deposition of cobalt ferrite nanoparticles. The increased contrast enabled visualization of the micro and nanosized particles inside the paper matrix using multiphoton microscopy, X-ray microtomography and SEM-EDX. The results indicate that fibrillary fines enrich at pores and fiber-fiber junctions, thereby increasing the relative bonded area between fibers to enhance paper strength while CNF seems to additionally form an inner 3D network.

Review: Periodate oxidation of wood polysaccharides-Modulation of hierarchies.

Periodate oxidation of polysaccharides has transitioned from structural analysis into a modification method for engineered materials. This review summarizes the research on this topic. Fibers, fibrils, crystals, and molecules originating from forests that have been subjected to periodate oxidation can be crosslinked with other entities via the generated aldehyde functionality, that can also be oxidized or reduced to carboxyl or alcohol functionality or used as a starting point for further modification. Periodate-oxidized materials can be subjected to thermal transitions that differ from the native cellulose. Oxidation of polysaccharides originating from forests often features oxidation of structures rather than liberated molecules. This leads to changes in macro, micro, and supramolecular assemblies and consequently to alterations in physical properties. This review focuses on these aspects of the modulation of structural hierarchies due to periodate oxidation.

The Exo-Polysaccharide Component of Extracellular Matrix is Essential for the Viscoelastic Properties of Bacillus subtilis Biofilms.

Bacteria are known to form biofilms on various surfaces. Biofilms are multicellular aggregates, held together by an extracellular matrix, which is composed of biological polymers. Three principal components of the biofilm matrix are exopolysaccharides (EPS), proteins, and nucleic acids. The biofilm matrix is essential for biofilms to remain organized under mechanical stress. Thanks to their polymeric nature, biofilms exhibit both elastic and viscous mechanical characteristics; therefore, an accurate mechanical description needs to take into account their viscoelastic nature. Their viscoelastic properties, including during their growth dynamics, are crucial for biofilm survival in many environments, particularly during infection processes. How changes in the composition of the biofilm matrix affect viscoelasticity has not been thoroughly investigated. In this study, we used interfacial rheology to study the contribution of the EPS component of the matrix to viscoelasticity of Bacillus subtilis biofilms. Two strategies were used to specifically deplete the EPS component of the biofilm matrix, namely (i) treatment with sub-lethal doses of vitamin C and (ii) seamless inactivation of the eps operon responsible for biosynthesis of the EPS. In both cases, the obtained results suggest that the EPS component of the matrix is essential for maintaining the viscoelastic properties of bacterial biofilms during their growth. If the EPS component of the matrix is depleted, the mechanical stability of biofilms is compromised and the biofilms become more susceptible to eradication by mechanical stress.