An Analytical Toolbox for Fast and Straightforward Structural Characterisation of Commercially Available Tannins.

Both condensed and hydrolysable tannins represent versatile natural polyphenolic structures exhibiting a broad range of activities that could be exploited in various fields including nutraceutics, cosmesis, consumer care, household and pharmaceutical applications. Various tannins are commercially available nowadays for use in such application fields. We have analysed a representative selection of commercially available condensed and hydrolysable tannins for structural features and purity. Using a combination of quantitative 31P NMR spectroscopy, HSQC measurements, MALDI-ToF analyses, gel permeation chromatography and wet chemical analysis, detailed structural characterisations and descriptions were possible, allowing for verification and falsification of claimed structural features.

Lignosulfonate Microcapsules for Delivery and Controlled Release of Thymol and Derivatives.

Thymol and the corresponding brominated derivatives constitute important biological active molecules as antibacterial, antioxidant, antifungal, and antiparasitic agents. However, their application is often limited, because their pronounced fragrance, their poor solubility in water, and their high volatility. The encapsulation of different thymol derivatives into biocompatible lignin-microcapsules is presented as a synergy-delivering remedy. The adoption of lignosulfonate as an encapsulating material possessing relevant antioxidant activity, as well as general biocompatibility allows for the development of new materials that are suitable for the application in various fields, especially cosmesis. To this purpose, lignin microcapsules containing thymol, 4-bromothymol, 2,4-dibromothymol, and the corresponding O-methylated derivatives have been efficiently prepared through a sustainable ultrasonication procedure. Actives could be efficiently encapsulated with efficiencies of up to 50%. To evaluate the applicability of such systems for topical purposes, controlled release experiments have been performed in acetate buffer at pH 5.4, to simulate skin pH: all of the capsules show a slow release of actives, which is strongly determined by their inherent lipophilicity.

Characterization of Eucalyptus nitens Lignins Obtained by Biorefinery Methods Based on Ionic Liquids.

Eucalyptus nitens wood samples were subjected to consecutive stages of hydrothermal processing for hemicellulose solubilization and delignification with an ionic liquid, i.e., either 1-butyl-3-methylimidazolium hydrogen sulfate or triethylammonium hydrogen sulfate. Delignification experiments were carried out a 170 C for 10-50 min. The solid phases from treatments, i.e., cellulose-enriched solids, were recovered by centrifugation, and lignin was separated from the ionic liquid by water precipitation. The best delignification conditions were identified on the basis of the results determined for delignification percentage, lignin recovery yield, and cellulose recovery in solid phase. The lignins obtained under selected conditions were characterized in deep by 31P-NMR, 13C-NMR, HSQC, and gel permeation chromatography. The major structural features of the lignins were discussed in comparison with the results determined for a model Ionosolv lignin.

Detailed Chemical Composition of Condensed Tannins via Quantitative (31)P NMR and HSQC Analyses: Acacia catechu, Schinopsis balansae, and Acacia mearnsii.

The chemical composition of Acacia catechu, Schinopsis balansae, and Acacia mearnsii proanthocyanidins has been determined using a novel analytical approach that rests on the concerted use of quantitative (31)P NMR and two-dimensional heteronuclear NMR spectroscopy. This approach has offered significant detailed information regarding the structure and purity of these complex and often elusive proanthocyanidins. More specifically, rings A, B, and C of their flavan-3-ol units show well-defined and resolved absorbance regions in both the quantitative (31)P NMR and HSQC spectra. By integrating each of these regions in the (31)P NMR spectra, it is possible to identify the oxygenation patterns of the flavan-3-ol units. At the same time it is possible to acquire a fingerprint of the proanthocyanidin sample and evaluate its purity via the HSQC information. This analytical approach is suitable for both the purified natural product proanthocyanidins and their commercial analogues. Overall, this effort demonstrates the power of the concerted use of these two NMR techniques for the structural elucidation of natural products containing labile hydroxy protons and a carbon framework that can be traced out via HSQC.

Modification of Kraft Lignin to Expose Diazobenzene Groups: Toward pH- and Light-Responsive Biobased Polymers.

A pH- and light-responsive polymer has been synthesized from softwood kraft lignin by a two-step strategy that aimed to incorporate diazobenzene groups. Initially, styrene oxide was reacted with the phenolic hydroxyl groups in lignin, to offer the attachment of benzene rings, thus creating unhindered reactive sites for further modifications. The use of advanced spectroscopic techniques ((1)H and (31)P NMR, UV and FTIR) demonstrated that the reaction was quantitative and selective toward the phenolic hydroxyl groups. In a second step, the newly incorporated benzene rings were reacted with a diazonium cation to form the target diazobenzene motif, whose formation was again thoroughly verified. As anticipated, the diazobenzene-containing kraft lignin derivatives showed a pH-dependent color change in solution and light-responsive properties resulting from the cis-trans photoisomerization of the diazobenzene group.

Ultrasound driven assembly of lignin into microcapsules for storage and delivery of hydrophobic molecules.

Oil-filled microcapsules of kraft lignin were synthesized by first creating an oil in water emulsion followed by a high-intensity, ultrasound-assisted cross-linking of lignin at the water/oil interface. The rationale behind our approach is based on promoting documented lignin hydrophobic interactions within the oil phase, followed by locking the resulting spherical microsystems by covalent cross-linking using a high intensity ultrasound treatment. As further evidence in support of our rationale, confocal and optical microscopies demonstrated the uniformly spherical morphology of the created lignin microparticles. The detailed elucidation of the cross-linking processes was carried out using gel permeation chromatography (GPC) and quantitative (31)P NMR analyses. The ability of lignin microcapsules to incorporate and release Coumarin-6 was evaluated in detail. In vitro studies and confocal laser scanning microscopy analysis were carried out to assess the internalization of capsules into Chinese hamster ovary (CHO) cells. This part of our work demonstrated that the lignin microcapsules are not cytotoxic and readily incorporated in the CHO cells.

The Laccase Catalysed Tandem Lignin Depolymerisation/Polymerisation.

The development of strategies allowing either the production of high value phenolics, or the isolation of properties-enhanced materials from technical lignins represents a fundamental step in the industrial upcycling of technical lignins. Both aims are met by the strategy presented in the present work, relying on the coupling of solvent-based fractionation with the oxidative action of a new type of alkaline-stable genetically modified bacterial laccase. The described approach succeeded in the tandem, high-yield and selective isolation of valuable lignin-monomeric compounds (MCs) and high molecular weight and hydrophobicity-tailored polymerised materials (PMs) from two technical lignins, namely softwood kraft lignin (SKL), and wheat straw organosolv lignin (WSL). With respect to MCs, higher yields as compared to similar studies (up to 17.2 mg/g) were achieved. PMs from SKL samples where characterised by an almost quadrupled Mw, while in the case of WSL the Mw was approximately doubled. Noteworthy, the reaction conditions were optimized in terms of reaction temperature, time, enzymatic loading, and alkalinity for the selective production of single MCs. Most interestingly, technical lignins as well as their fractions and the PMs deriving from their laccase-catalysed oxidation showed increased hydrophobicity.

Kraft (Nano)Lignin as Reactive Additive in Epoxy Polymer Bio-Composites.

The demand for high-performance bio-based materials towards achieving more sustainable manufacturing and circular economy models is growing significantly. Kraft lignin (KL) is an abundant and highly functional aromatic/phenolic biopolymer, being the main side product of the pulp and paper industry, as well as of the more recent 2nd generation biorefineries. In this study, KL was incorporated into a glassy epoxy system based on the diglycidyl ether of bisphenol A (DGEBA) and an amine curing agent (Jeffamine D-230), being utilized as partial replacement of the curing agent and the DGEBA prepolymer or as a reactive additive. A D-230 replacement by pristine (unmodified) KL of up to 14 wt.% was achieved while KL-epoxy composites with up to 30 wt.% KL exhibited similar thermo-mechanical properties and substantially enhanced antioxidant properties compared to the neat epoxy polymer. Additionally, the effect of the KL particle size was investigated. Ball-milled kraft lignin (BMKL, 10 µm) and nano-lignin (NLH, 220 nm) were, respectively, obtained after ball milling and ultrasonication and were studied as additives in the same epoxy system. Significantly improved dispersion and thermo-mechanical properties were obtained, mainly with nano-lignin, which exhibited fully transparent lignin-epoxy composites with higher tensile strength, storage modulus and glass transition temperature, even at 30 wt.% loadings. Lastly, KL lignin was glycidylized (GKL) and utilized as a bio-based epoxy prepolymer, achieving up to 38 wt.% replacement of fossil-based DGEBA. The GKL composites exhibited improved thermo-mechanical properties and transparency. All lignins were extensively characterized using NMR, TGA, GPC, and DLS techniques to correlate and justify the epoxy polymer characterization results.

Lignin-derived Sustainable Nano-platforms: A Multifunctional Solution for an Efficient Dye Removal.

In contrast to conventional non-biobased adsorbents, lignin emerges as a cost-effective and environmentally benign alternative for water treatment. This study identifies unexpected and unpredicted multifunctional properties of lignin nanoparticles (LNPs). LNPs, which are prepared by simple physical processes, demonstrated for the first time to behave as multifunctional materials able to adsorb and photodegrade methylene blue (MB) in aqueous medium upon UV irradiation. Furthermore, the synthetic approach adopted to synthesize LNPs - and therefore their surface properties - strongly affects their performances. More specifically, LNPs obtained by solvent-antisolvent nanoprecipitation (SLNPs) show the highest MB adsorption properties (98% removal), reaching a maximum adsorption capacity of 43 mg g-1, and the fastest adsorption kinetics with respect to other lignin-based adsorbents. Conversely, hydrotropic LNPs (HLNPs) exhibit exceptional photocatalytic activity, resulting in 98% MB degradation over 6 hours of UV irradiation, combined with the ability to be easily recycled and reused.  The present effort paves the way for the use of LNPs as efficient multifunctional materials able to perform concurrently adsorption and photocatalytic degradation of dye pollutants, toward the creation of a sustainable biobased water treatment platform.

Kraft Lignin: A Valuable, Sustainable Resource, Opportunities and Challenges.

Kraft lignin, a by-product from the production of pulp, is currently incinerated in the recovery boiler during the chemical recovery cycle, generating valuable bioenergy and recycling inorganic chemicals to the pulping process operation. Removing lignin from the black liquor or its gasification lowers the recovery boiler load enabling increased pulp production. During the past ten years, lignin separation technologies have emerged and the interest of the research community to valorize this underutilized resource has been invigorated. The aim of this Review is to give (1) a dedicated overview of the kraft process with a focus on the lignin, (2) an overview of applications that are being developed, and (3) a techno-economic and life cycle asseeements of value chains from black liquor to different products. Overall, it is anticipated that this effort will inspire further work for developing and using kraft lignin as a commodity raw material for new applications undeniably promoting pivotal global sustainability concerns.

Layer-by-Layer coated tyrosinase: An efficient and selective synthesis of catechols.

Agaricus bisporous tyrosinase was immobilized on commercial available epoxy-resin Eupergit®C250L and then coated by the Layer-by-Layer method (LbL). The two novel heterogeneous biocatalysts were characterized for their morphology, pH and storage stability, kinetic properties (K(m), V(max), V(max)/K(m)) and reusability. These biocatalysts were used for the efficient and selective synthesis of bioactive catechols under mild and environmental friendly experimental conditions. Ascorbic acid was added in the reaction medium to inhibit the formation of ortho-quinones, thus avoiding the known enzyme suicide inactivation process. Catechols were obtained mostly in quantitative yields and conversion of substrate. Tyrosinase immobilized on Eupergit®C250L and coated by the LbL method showed better catalytic activities, higher pH and storage stability, and reusability with respect to immobilized uncoated tyrosinase. Since chemical procedures to synthesize catechols are often expensive and with high environmental impact, the use of immobilized tyrosinase represents an efficient alternative for the preparation of this family of bioactive compounds.

Simple Strategies to Modulate the pH-Responsiveness of Lignosulfonate-Based Delivery Systems.

The extensive use of non-degradable microplastics in a wide plethora of daily life products is causing serious pollution problems. More ecofriendly solutions are therefore urgently needed. In this context, the use of lignin, a largely available aromatic polymer, may represent a viable option. Due to the self-assembly ability of its molecules, lignin is in fact an ideal matrix for the fabrication of nanostructures. In this study, lignosulfonate microcapsules containing a limonene core were prepared and characterized in terms of their dimensions and of the physicochemical characteristics of the capsule-forming lignosulfonate molecules. The main purpose is to elucidate the key properties governing the pH-responsive behavior of the capsules to be able to achieve better control over the release kinetics of the entrapped compound(s). The results demonstrate that both the molecular weight and the concentration of sulfonate groups are the most important factors in this respect. Based on these findings, two strategies were followed to further tailor the capsules' behavior: (i) fractionation of the starting lignosulfonate by solvent extraction and (ii) introduction of a specific additive in the formulation. The first approach permitted to fabricate highly resistant capsules both in acidic, as well as in alkaline conditions, while in the second case the chemical structure of the additive, the diester diveratryl sebacate, allowed for fast kinetics of release, as values above 70% were reached after 24 h of incubation at pH 4 and pH 12.

The Laccase-Lig Multienzymatic Multistep System in Lignin Valorization.

Invited for this month's cover is the work by Claudia Crestini and collaborators at Ca'Foscari University of Venice, Italy, and University of Insubria, Italy. The image shows the formation of low-molecular-weight compounds by the oxidative depolymerization of lignin by the laccase-Lig multienzymatic multistep system. The Research Article itself is available at 10.1002/cssc.202201147.

The Laccase-Lig Multienzymatic Multistep System in Lignin Valorization.

A laccase-Lig multienzymatic multistep system for lignin depolymerization was designed and developed. Studies were performed on pristine and fractionated lignins (Kraft and Organosolv) using a specific cascade of enzymes, that is, laccases from Bacillus licheniformis and from Funalia trogii, respectively for Kraft and Organosolv lignin, followed by the Lig system from Sphingobium sp. SYK-6 (ß-etherases Lig E and Lig F, glutathione lyase Lig G). Careful elucidation of the structural modifications occurring in the residual lignins associated with the identification and quantification of the generated low-molecular-weight compounds showed that (i) the laccase-Lig system cleaves non-phenolic aryl glycerol ß-O-4 aryl ether bonds, and (ii) the overall reactivity is heavily dependent on the individual lignin structure. More specifically, samples with low phenolic/aliphatic OH groups ratio undergo net depolymerization, while an increased phenolic/aliphatic OH ratio results in the polymerization of the residual lignin irrespective of its botanical origin and isolation process.

Effect of chemical modifications of tannins on their antimicrobial and antibiofilm effect against Gram-negative and Gram-positive bacteria.

Background: Tannins have demonstrated antibacterial and antibiofilm activity, but there are still unknown aspects on how the chemical properties of tannins affect their biological properties. We are interested in understanding how to modulate the antibiofilm activity of tannins and in delineating the relationship between chemical determinants and antibiofilm activity. Materials and methods: The effect of five different naturally acquired tannins and their chemical derivatives on biofilm formation and planktonic growth of Salmonella Typhimurium, Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus was determined in the Calgary biofilm device. Results: Most of the unmodified tannins exhibited specific antibiofilm activity against the assayed bacteria. The chemical modifications were found to alter the antibiofilm activity level and spectrum of the tannins. A positive charge introduced by derivatization with higher amounts of ammonium groups shifted the anti-biofilm spectrum toward Gram-negative bacteria, and derivatization with lower amounts of ammonium groups and acidifying derivatization shifted the spectrum toward Gram-positive bacteria. Furthermore, the quantity of phenolic OH-groups per molecule was found to have a weak impact on the anti-biofilm activity of the tannins. Conclusion: We were able to modulate the antibiofilm activity of several tannins by specific chemical modifications, providing a first approach for fine tuning of their activity and antibacterial spectrum.

Lignin-based nano-enabled agriculture: A mini-review.

Nowadays sustainable nanotechnological strategies to improve the efficiency of conventional agricultural practices are of utmost importance. As a matter of fact, the increasing use of productive factors in response to the growing food demand plays an important role in determining the environmental impact of agriculture. In this respect, low-efficiency conventional practices are becoming obsolete. On the other hand, the exploitation of nanoscaled systems for the controlled delivery of fertilizers, pesticides and herbicides shows great potential towards the development of sustainable, efficient and resilient agricultural processes, while promoting food security. In this context, lignin - especially in the form of its nanostructures - can play an important role as sustainable biomaterial for nano-enabled agricultural applications. In this review, we present and discuss the current advancements in the preparation of lignin nanoparticles for the controlled release of pesticides, herbicides, and fertilizers, as well as the latest findings in terms of plant response to their application. Special attention has been paid to the state-of-the-art literature concerning the release performance of these lignin-based nanomaterials, whose efficiency is compared with the conventional approaches. Finally, the major challenges and the future scenarios of lignin-based nano-enabled agriculture are considered.

N-Doped Carbon Dot Hydrogels from Brewing Waste for Photocatalytic Wastewater Treatment.

The brewery industry annually produces huge amounts of byproducts that represent an underutilized, yet valuable, source of biobased compounds. In this contribution, the two major beer wastes, that is, spent grains and spent yeasts, have been transformed into carbon dots (CDs) by a simple, scalable, and ecofriendly hydrothermal approach. The prepared CDs have been characterized from the chemical, morphological, and optical points of view, highlighting a high level of N-doping, because of the chemical composition of the starting material rich in proteins, photoluminescence emission centered at 420 nm, and lifetime in the range of 5.5-7.5 ns. With the aim of producing a reusable catalytic system for wastewater treatment, CDs have been entrapped into a polyvinyl alcohol matrix and tested for their dye removal ability. The results demonstrate that methylene blue can be efficiently adsorbed from water solutions into the composite hydrogel and subsequently fully degraded by UV irradiation.

Elucidation of lignin structure by quantitative 2D NMR.

Quick quantitative HSQC (QQ-HSQC) was applied to quantitative evaluation of different inter-unit linkages in an array of milled softwood and hardwood and technical lignins by using the guaiacyl C2 and syringyl C2-C6 signals as internal standards. The results were found to be highly reproducible and comparable with earlier literature reports. The advantage of QQ-HSQC NMR analysis of lignin is contemporary detection and quantification of lignin inter-unit linkages with a direct, non-destructive method requiring short acquisition times.