Phototunable Lignin Plastics to Enable Recyclability.

The accumulation of non-degradable petrochemical plastics imposes a significant threat to the environment and ecosystems. We addressed this challenge by designing a new type of phototunable plastics based on the unique lignin chemistry to enable readily end-life recycling. The advanced material design leveraged the efficient photocatalytic lignin depolymerization by ZnO nanoparticles to build lignin-polymethyl methacrylate (PMMA)-ZnO blends. We first demonstrated the highly effective phototunable lignin depolymerization in the complex polymer blend matrix and explored the molecular mechanisms. The technical barriers of mechanical property and recycling processing were then addressed by a new blend design with lignin core grafted with PMMA polymer. The new process has resulted in a new type of PMMA-g-lignin blend, which significantly improved the mechanical properties, making it comparable to PMMA alone. More importantly, the mechanical properties of the UV-treated blend decreased drastically in the new design, whereas the properties did not reduce in the non-grafted blends upon UV exposure. The results highlighted that the new blend design based on graftization maximized the impact of lignin depolymerization on blend structure and recyclability. Based on the results, we developed a process integrating UV and alkaline treatments to recycle PMMA for plastics and fractionated lignin for bioconversion or other applications in the new phototunable plastics.

Transforming biorefinery designs with 'Plug-In Processes of Lignin' to enable economic waste valorization.

Biological lignin valorization has emerged as a major solution for sustainable and cost-effective biorefineries. However, current biorefineries yield lignin with inadequate fractionation for bioconversion, yet substantial changes of these biorefinery designs to focus on lignin could jeopardize carbohydrate efficiency and increase capital costs. We resolve the dilemma by designing 'plug-in processes of lignin' with the integration of leading pretreatment technologies. Substantial improvement of lignin bioconversion and synergistic enhancement of carbohydrate processing are achieved by solubilizing lignin via lowering molecular weight and increasing hydrophilic groups, addressing the dilemma of lignin- or carbohydrate-first scenarios. The plug-in processes of lignin could enable minimum polyhydroxyalkanoate selling price at as low as $6.18/kg. The results highlight the potential to achieve commercial production of polyhydroxyalkanoates as a co-product of cellulosic ethanol. Here, we show that the plug-in processes of lignin could transform biorefinery design toward sustainability by promoting carbon efficiency and optimizing the total capital cost.

Enhancement of polyhydroxyalkanoate production by co-feeding lignin derivatives with glycerol in Pseudomonas putida KT2440.

BACKGROUND: Efficient utilization of all available carbons from lignocellulosic biomass is critical for economic efficiency of a bioconversion process to produce renewable bioproducts. However, the metabolic responses that enable Pseudomonas putida to utilize mixed carbon sources to generate reducing power and polyhydroxyalkanoate (PHA) remain unclear. Previous research has mainly focused on different fermentation strategies, including the sequential feeding of xylose as the growth stage substrate and octanoic acid as the PHA-producing substrate, feeding glycerol as the sole carbon substrate, and co-feeding of lignin and glucose. This study developed a new strategy-co-feeding glycerol and lignin derivatives such as benzoate, vanillin, and vanillic acid in Pseudomonas putida KT2440-for the first time, which simultaneously improved both cell biomass and PHA production. RESULTS: Co-feeding lignin derivatives (i.e. benzoate, vanillin, and vanillic acid) and glycerol to P. putida KT2440 was shown for the first time to simultaneously increase cell dry weight (CDW) by 9.4-16.1% and PHA content by 29.0-63.2%, respectively, compared with feeding glycerol alone. GC-MS results revealed that the addition of lignin derivatives to glycerol decreased the distribution of long-chain monomers (C10 and C12) by 0.4-4.4% and increased the distribution of short-chain monomers (C6 and C8) by 0.8-3.5%. The 1H-13C HMBC, 1H-13C HSQC, and 1H-1H COSY NMR analysis confirmed that the PHA monomers (C6-C14) were produced when glycerol was fed to the bacteria alone or together with lignin derivatives. Moreover, investigation of the glycerol/benzoate/nitrogen ratios showed that benzoate acted as an independent factor in PHA synthesis. Furthermore, 1H, 13C and 31P NMR metabolite analysis and mass spectrometry-based quantitative proteomics measurements suggested that the addition of benzoate stimulated oxidative-stress responses, enhanced glycerol consumption, and altered the intracellular NAD+/NADH and NADPH/NADP+ ratios by up-regulating the proteins involved in energy generation and storage processes, including the Entner-Doudoroff (ED) pathway, the reductive TCA route, trehalose degradation, fatty acid ß-oxidation, and PHA biosynthesis. CONCLUSIONS: This work demonstrated an effective co-carbon feeding strategy to improve PHA content/yield and convert lignin derivatives into value-added products in P. putida KT2440. Co-feeding lignin break-down products with other carbon sources, such as glycerol, has been demonstrated as an efficient way to utilize biomass to increase PHA production in P. putida KT2440. Moreover, the involvement of aromatic degradation favours further lignin utilization, and the combination of proteomics and metabolomics with NMR sheds light on the metabolic and regulatory mechanisms for cellular redox balance and potential genetic targets for a higher biomass carbon conversion efficiency.

Stereolithography 3D Printing of Lignin-Reinforced Composites with Enhanced Mechanical Properties.

Due to the availability, biodegradability, and biological effects, lignin has emerged as an interesting alternative to petroleum-based compounds for developing sustainable chemicals, materials, and composites. In this study, lignin at various concentrations was incorporated into methacrylate resin via solution blending to fabricate lignin-reinforced composites using stereolithography apparatus three-dimensional printing. Softwood kraft lignin in the amounts of 0.2, 0.4, 0.5, 0.8, and 1.0 wt % in the methacrylate resin was used as a printing ink, and the gel contents and relative contents of the residual resin in the printed samples with various lignin concentrations were measured. The effects of the lignin on the ultimate mechanical properties of the non-postcured and postcured printed composites were determined. The tensile testing results revealed that the incorporation of lignin in the composite increased the tensile strength by 46-64% and Young's modulus by 13-37% for the postcured printed composites compared with that of the control sample (no lignin added). Employing a 0.4 wt % softwood kraft lignin, the tensile strength of the postcured printed composite reached the highest value of 49.0 MPa, which was a 60% increase in comparison to that of the control sample with 30.7 MPa. Scanning electron microscopy images of the fracture samples illustrated that the lignin-incorporated composites exhibited a rougher fracture surface that can presumably dissipate the stress, which could be a contributing factor for the mechanical enhancement.

Determination of hydroxyl groups in biorefinery resources via quantitative 31P NMR spectroscopy.

The analysis of chemical structural characteristics of biorefinery product streams (such as lignin and tannin) has advanced substantially over the past decade, with traditional wet-chemical techniques being replaced or supplemented by NMR methodologies. Quantitative 31P NMR spectroscopy is a promising technique for the analysis of hydroxyl groups because of its unique characterization capability and broad potential applicability across the biorefinery research community. This protocol describes procedures for (i) the preparation/solubilization of lignin and tannin, (ii) the phosphitylation of their hydroxyl groups, (iii) NMR acquisition details, and (iv) the ensuing data analyses and means to precisely calculate the content of the different types of hydroxyl groups. Compared with traditional wet-chemical techniques, the technique of quantitative 31P NMR spectroscopy offers unique advantages in measuring hydroxyl groups in a single spectrum with high signal resolution. The method provides complete quantitative information about the hydroxyl groups with small amounts of sample (~30 mg) within a relatively short experimental time (~30-120 min).

Investigating the correlation of biomass recalcitrance with pyrolysis oil using poplar as the feedstock.

Pyrolysis of five poplar samples with differing degrees of recalcitrance was performed; the correlations between the poplar enzymatic hydrolysis glucose yields and the physicochemical properties of pyrolysis product were investigated in this study. Sugar release of five poplar samples varied from 48.1 to 112.3 mg/g for glucose, and 12.0 to 32.4 mg/g for xylose. The yield of pyrolysis products was calculated and the molecular weight distribution of pyrolysis oils was measured by GPC, ranging from 268 to 289 g/mol for its weight-average molecular weight. GC-MS analysis of the bio-oil exhibited a strong correlation between biomass recalcitrance and guaiacyl-type structures in bio-oils. The correlation between biomass recalcitrance and the ratio of syringyl-to-guaiacyl-type-related structures was also assessed. The results from quantitative 31P NMR indicated some correlation between biomass recalcitrance and the guaiacyl hydroxyl groups in bio-oils. These results illustrate correlations and differences between converting biomass to biofuels via the biological and thermal platform.

One-pot transformation of lignocellulosic biomass into crude bio-oil with metal chlorides via hydrothermal and supercritical ethanol processing.

Grape seeds were deconstructed in both hydrothermal and supercritical ethanol media with a combination of two metal chlorides (TiCl4:MgCl2) to produce bio-oils. The use of metal chloride additives in supercritical ethanol achieved the highest bio-oil yield of 49.2 wt% (300 °C, 30 min). Both the hydrothermal and supercritical ethanol deconstruction with the additives (TiCl4:MgCl2 = 4 mmol:4mmol) produced the bio-oils with a higher heating value (HHV) of 35 MJ/Kg. Gas chromatography-mass spectrometry (GC-MS) analysis of the bio-oils showed that the major products in bio-oils from the hydrothermal deconstruction were acids while the majority products in bio-oils form the supercritical ethanol deconstruction were esters. Nuclear magnetic resonance (NMR) data of the bio-oils suggested that both hydrothermal and supercritical ethanol deconstruction with metal chlorides significantly reduced the non-condensed OH and oxygenated lignin sub-units in bio-oils; while only supercritical ethanol deconstruction with metal chlorides reduced the aliphatic OH and O-alkylated structures in bio-oils.

Cross-linked poly(methyl vinyl ether-co-maleic acid)/poly(ethylene glycol)/nanocellulosics foams via directional freezing.

Aligned micro- and nanoporous materials have gained tremendous interest since they provide great potential in organic electronics, absorbents, biomedicine and tissue engineering. Herein, cellulose nanofibrils (CNFs) and cellulose nanocrystals (CNCs) cross-linked with poly(methyl vinyl ether-co-maleic acid) (PMVEMA) and poly(ethylene glycol) (PEG) foams were prepared by both the directional and un-directional freezing techniques. Aligned three-dimensional porous structures were observed for the directional frozen foams after lyophilization via scanning electron microscope (SEM). As a result, these highly organized structures exhibited enhanced mechanical performance properties. Particularly, for the 25% CNF foams, the compression modulus increased 60% compared with the un-directional frozen samples. These nanocellulosics-based foams could absorb up to 10-fold water of their initial weight with excellent water stability when immersed in water for more than 48 h. Overall, this study describes a novel process combining cross-linking and directional freezing which successfully fabricates naturally derived foams with anisotropic structure.

Biomimetic composite scaffold from an in situ hydroxyapatite coating on cellulose nanocrystals.

A novel nanocomposite scaffold was developed by homogeneous deposition of hydroxyapatite (HAP) on a cellulose nanocrystals (CNCs) matrix suspended in a simulated body fluid (SBF). By adjusting the pH of the SBF, the HAP content in the nanocomposite could be controlled between 15 wt% and 47 wt%. Physical and chemical characteristics of the nanocomposites were analyzed by SEM, FTIR, XRD, SAED, and TEM, which confirmed the successful incorporation of HAP onto the CNCs. The nanocomposites were then freeze-casted into porous scaffolds by different solidification technologies (i.e., directional freezing (DF), plunging in liquid N2 (PL) or in a -20 °C freezer (FZ)) followed by lyophilization. Compression testing of the HAP/CNCs foams indicated that DF caused significant improvement in mechanical properties due to the specific orientation and anisotropic porous structure compared to conventional freezing methods such as PL and FZ. Moreover, the scaffold with high HAP content exhibited improved mechanical and thermal properties, which holds potential for application in bone tissue engineering.

Kinetic understanding of nitrogen supply condition on biosynthesis of polyhydroxyalkanoate from benzoate by Pseudomonas putida KT2440.

Nitrogen supply is critical to the synthesis of intracellular PHA in various bacteria. However, the specific role of the nitrogen in synthesizing PHA from benzoate, a lignin model compound use for the study of bacteria catabolism of aromatics, is still not clear. In this study, two culture conditions were maintained for Pseudomonas putida KT2440 to produce PHA using benzoate as a carbon source. Under nitrogen-limited and surplus conditions, the accumulation of PHA was to 37.3% and 0.25% of cell dry weight, respectively. A model fit to the kinetics of biomass growth and PHA accumulation showed good agreement with data. GC-MS and NMR showed that PHA contained six hydroxyl fatty acid monomers under nitrogen-limited conditions, while two monomers were identified under nitrogen surplus conditions. The average molecular weight of PHA increased after the nitrogen source was exhausted. These results provide a promising strategy for optimization of lignin to PHA yields.

Ethanol: A Promising Green Solvent for the Deconstruction of Lignocellulose.

Growing energy demand, environmental impact, energy security issues, and rural economic development have encouraged the development of sustainable renewable fuels. Nonfood lignocellulosic biomass is a suitable source for sustainable energy because the biomass feedstocks are low cost, abundant, and carbon neutral. Recent thermochemical conversion studies are frequently directed at converting biomass into high-quality liquid fuel precursors or chemicals in a single step. Supercritical ethanol has been selected as a promising solvent medium to deconstruct lignocellulosic biomass because ethanol has extraordinary solubility towards lignocellulosic biomass and can be resourced from cellulosic ethanol facilities. This review provides a critical insight into both catalytic and noncatalytic strategies of lignocellulose deconstruction. In this context, the supercritical ethanol deconstruction pathways are thoroughly reviewed; GC-MS, 1D and 2D NMR spectroscopy, and elemental analysis strategies towards liquid biomass deconstruction products are also critically presented. This review aims to provide readers a broad and accurate roadmap of novel biomass to biofuel conversion techniques.

Laccase-mediated functionalization of chitosan with 4-hexyloxyphenol enhances antioxidant and hydrophobic properties of copolymer.

An effective method to functionalize chitosan with 4-hexyloxyphenol (HP) under homogeneous reaction conditions was developed using laccase as the catalyst. The resulting copolymer was characterized for chemical structure, grafted-HP content, surface morphology, thermal stability, antioxidant capacity, hydrophobic properties and tensile strength. Solid-state 13C NMR spectrum confirmed the incorporation of HP onto chitosan. X-ray diffraction (XRD) showed a decrease in the degree of crystallinity for laccase/HP treated chitosan compared to pure chitosan. The grafted-HP content in laccase/HP-treated chitosan first increased and then declined with increase of the initial HP/chitosan ratio. A heterogeneous surface with spherical particles on the laccase/HP treated chitosan was observed by environmental scanning electron microscopy (ESEM) and scanning probe microscopy (SPM). The laccase/HP treatment of chitosan improved the thermal stability of copolymer. More significantly, the HP functionalized chitosan showed greatly improved ABTS+ and DPPH radicals scavenging capacity, compared with pure chitosan. The hydrophobicity property of the HP functionalized chitosan also significantly increased although its tensile strength decreased. This new type of composite with double functionalities (i.e., antioxidant and hydrophobic) could potentially be used as food packaging materials or coating agents.

Characterization of products from hydrothermal carbonization of pine.

This study aims to reveal the structural features and reaction pathways for solid-liquid products from hydrothermal carbonization of Loblolly pine, where the solid products can be used as catalysts, adsorbents and electrode materials while liquid products can be treated yielding fuels and platform chemicals. Results revealed when treated at 240°C, cellulose and hemicellulose were degraded, in part, to 5-hydroxy-methyl furfural and furfural which were further transformed to aromatic structures via ring opening and Diels Alder reactions. Lignin degradation and formation of carbon-carbon bonds, forming aromatic motifs in the presence of furanic compounds connected via aliphatic bridges, ether or condensation reactions. After hydrothermal treatment, condensed aromatic carbon materials with methoxy groups were recovered with high fixed carbon content and HHV. The recovered liquid products are lignin-like value-added chemicals consisting of furfural and polyaromatic structure with alkanes and carboxyl, their total hydroxyl group content decreased when increasing reaction time.

A bright red fluorescent cyanine dye for live-cell nucleic acid imaging, with high photostability and a large Stokes shift.

Many probes for nucleic acids are available, but few of them satisfy multiple criteria, particularly high photostability to endure laser scanning. We report a cyanine dye TO3-CN for the first time, synthesised by introducing a CN group to the trimethine chain of the classical red emitting TO-3 dye to improve its photostability, as well as their spectral properties and interaction with nucleic acids. TO3-CN shows excellent light fastness and a large fluorescence Stokes shift (more than 40 nm). Because of its sensitive fluorescence response to nucleic acids with a large fluorescence quantum yield (more than 0.7) and low cytotoxicity, this dye may be a potential candidate for nucleic acid detection in vitro and intracellular fluorescence imaging.