This study established a cost-efficient synthesis protocol for fabricating nitrogen-doped graphene-like carbons from biorefinery-derived lignin waste. The resultant carbon materials integrated precisely designed ultrathin two-dimensional layered configurations with extensive surface areas and mesoscale porous attributes. Utilizing TEM and in-situ TEM characterization techniques, we observed the dynamic derivation process of lignin-derived reactions and explored the controllable synthesis parameters. The rationally designed structures, abundant electrochemically active sites, enhanced electronic properties due to nitrogen doping, and efficient mass transfer and diffusion all contributed to the remarkable electrochemical activity and stability exhibited by the ligninderived carbon materials. As anode material for lithium storage, the lignin-derived carbon exhibited a high capacity retention ratio of 84.3 % after 500 cycles. For applications in fuel cell, the current density of ligninderived carbon catalysts remains as high as 92.0 % compared to the value of commercial Pt/C catalyst of 42.9 % after 30000 s' testing. Our experimental results suggest that these graphene-like carbon materials have great application potential as catalyst supports in hydrogen fuel cells and as electrode materials for lithium-ion storage.

5-Hydroxymethylfurfural (HMF) is a central platform molecule linking plant biomass to renewable fuels and chemicals, yet its scalable production is hindered by instability and costly separation. Here we present a temperature-responsive, phase-switchable acetone/betaine hydrochloride (BHC) aqueous system that unifies catalysis, product stabilization, and catalyst/solvent recycling in one platform. The system behaves as a liquid-liquid biphasic medium during fructose dehydration, enabling in situ extraction of HMF into acetone, and switches to a solid-liquid state upon cooling, allowing quantitative BHC recovery. Molecular simulations reveal that BHC accelerates dehydration via directional hydrogen bonding, while acetone selectively solvates HMF to suppress degradation. The system achieves up to 86.7% HMF yield at a high concentration of 30 wt% fructose, sustains >75% yield at 60 wt%, and remains robust over multiple cycles. A 1000-fold scale-up validates kilogram-scale production, and techno-economic analysis projects cost competitiveness with all reported routes. This phase-switchable concept establishes a broadly applicable catalytic strategy for catalyst/solvent recovery and product stabilization in biomass valorization.

A new and sustainable liquid organic hydrogen carrier, Lignin Jet Fuel-based Liquid Organic Hydrogen Carrier (LJF-HyC), has been discovered. This innovative LOHC is created from Lignin Jet Fuel (LJF) through dehydrogenation reactions. The process was carried out in situ using platinum nanoparticles supported on zeolite, resulting in a significant increase in aromatic carbon content. This increase indicates the successful formation of aromatic rings via C-H dissociation. In-situ Nuclear Magnetic Resonance (NMR) and gas chromatographic analyses revealed the formation of unsaturated and partially unsaturated compounds, including alkylbenzenes, tetralins, naphthalenes with double bond equivalence of 4-8, from six apparent reaction pathways, four of which can be major. The original LJF, consisting primarily of mono-, di-, and tricyclohexylalkanes (96 wt%), was converted to dehydrogenated products, constituting approximately 18.5 wt% of the LJF composition. These findings pave the way for developing sustainable hydrogen carriers derived from sustainable aviation fuels.

Fungi are vital to the bioeconomy, serving as key producers of food, beverages, biofuels, and medicines, while also acting as essential resource recyclers in ecosystem management. For nearly a century, oleaginous yeast and filamentous fungi have been explored for their proficiency in oleochemicals production and carbon storage. Lipogenesis is one of the most well-studied fungal processes, with substantial progress having been made through reductionist biochemical approaches; however, the physiology and metabolism of fungal systems operating under different conditions arise from the functions of thousands of proteins, for which very little is known outside of model yeast. In this review, we discuss how proteomics provides a valuable analytical approach to contextualize lipogenesis within a complex biological system, where lipid accumulation is fundamentally governed by changes in proteins of multiple pathways. In the past two decades, proteomics has been applied to study stress response to nutrient limitations, metabolism of various carbon and nitrogen sources, the lipid droplet hub of carbon storage, protein post-translational modifications and signaling pathways, as well as oleochemical biosynthesis, thereby advancing our understanding of the oleaginous phenotype. Over 40 studies are reviewed herein to evaluate the impact, critically assess the utility, and propose future applications of proteomics. In the coming years, large systems-level proteomics studies will lay a foundation for marrying modeling and metabolic engineering strategies to optimize oleochemicals production in oleaginous fungi.

The study aimed to examine the effects of adding biomass ash on the biochemical processes involved in fermentable sugar production. Corn stover was pretreated using several methods—hot water, dilute acid, alkaline, γ-valerolactone, and ionic liquid methods, each examined with ash loadings of 7.18

Oleaginous yeast are prodigious producers of oleochemicals, offering alternative and secure sources for applications in foodstuff, skincare, biofuels, and bioplastics. Nitrogen starvation is the primary strategy used to induce oil accumulation in oleaginous yeast as part of a global stress response. While research has demonstrated that post-translational modifications (PTMs), including phosphorylation and protein cysteine thiol oxidation (redox PTMs), are involved in signaling pathways that regulate stress responses in metazoa and algae, their role in oleaginous yeast remain understudied and unexplored. Towards linking the yeast oleaginous phenotype to protein function, we integrated lipidomics, redox proteomics, and phosphoproteomics to investigate Rhodotorula toruloides under nitrogen-rich and starved conditions over time. Our lipidomics results unearthed interactions involving sphingolipids and cardiolipins with ER stress and mitophagy. Our redox and phosphoproteomics data highlighted the roles of the AMPK, TOR, and calcium signaling pathways in regulation of lipogenesis, autophagy, and oxidative stress response. As a first, we also demonstrated that lipogenic enzymes including fatty acid synthase are modified as a consequence of shifts in cellular redox states due to nutrient availability. We conclude that lipid accumulation is largely a consequence of carbon rerouting and autophagy governed by changes to PTMs, and not increases in the abundance of enzymes involved in central carbon metabolism and fatty acid biosynthesis. Our systems-level approach sets the stage for acquiring multidimensional data sets for protein structural modeling and predicting the functional relevance of PTMs using Artificial Intelligence/Machine Learning (AI/ML). Coupled to those bioinformatics approaches, the putative PTM switches that we delineate will enable advanced metabolic engineering strategies to decouple lipid accumulation from nitrogen limitation.

Lignin depolymerization is crucial for producing fuels and chemicals. Catalytic hydrodeoxygenation offers a distinct method for lignin depolymerization, leading to improved bio-oil conversion yields. This approach involves breaking C-O bonds, hydrogenating rings, and opening rings. In this study, Ru-based bifunctional catalysts with a low Ru metal loading of 0.2 to 0.5 wt.

Catechyl lignin (C-lignin), a type of lignin found in vanilla seed coats and some members of the Cactaceae and Euphorbiaceae families, has gained significant attention. C-lignin possesses a homogeneous linear structure that is characterized by the presence of a catechol group comprising a benzene ring with two hydroxyl groups. C-lignin also has substructures according to specific linkage patterns between its constituent monolignols, i.e., coniferyl (C), hydroxyphenyl (H), guaiacyl (G), and sinapyl (S) monomers. The linear structure of C-lignin makes it an ideal source for the development of carbon fiber-based composites, whereas its nonetherified structure and low molecular weight favor its microbial conversion to various useful products. However, to fully realize the potential of C-lignin, it is important to obtain its significant quantities through gene regulation. Technology must be developed to address these challenges and achieve the goals successfully. Genetic engineering techniques have been developed to increase C-lignin accumulation in specific plants for valorization. The extraction of C-lignin from biomass materials involves various effective methods to depolymerize it, producing aromatic compounds like propyl and propenyl catechol. This makes C-lignin a promising material for depolymerization and unlocking its valuable use, thanks to its homogeneous catechyl units. This review explores the biochemical and molecular regulation of C-lignin biosynthesis. It discusses the role of various enzymes, genes, and regulators in the accumulation of C-lignin in different plant species. The review also delves into the techniques, catalysts, and solvents employed in the extraction of C-lignin. Additionally, it discusses the depolymerization of C-lignin to produce various aromatic compounds, as well as its application in developing carbon fibers and polymeric composites. Finally, the review highlights the challenges and potential opportunities associated with utilizing the homogeneous and linear structure of C-lignin for future applications.

Studying regulation of protein function at a systems level necessitates an understanding of the interplay among diverse posttranslational modifications (PTMs). A variety of proteomics sample processing workflows are currently used to study specific PTMs but rarely characterize multiple types of PTMs from the same sample inputs. Method incompatibilities and laborious sample preparation steps complicate large-scale physiological investigations and can lead to variations in results. The single-pot, solid-phase-enhanced sample preparation (SP3) method for sample cleanup is compatible with different lysis buffers and amenable to automation, making it attractive for high-throughput multi-PTM profiling. Herein, we describe an integrative SP3 workflow for multiplexed quantification of protein abundance, cysteine thiol oxidation, phosphorylation, and acetylation. The broad applicability of this approach is demonstrated using cell and tissue samples, and its utility for studying interacting regulatory networks is highlighted in a time-course experiment of cytokine-treated β-cells. We observed a swift response in the global regulation of protein abundances consistent with rapid activation of JAK-STAT and NF-κB signaling pathways. Regulators of these pathways as well as proteins involved in their target processes displayed multi-PTM dynamics indicative of complex cellular response stages: acute, adaptation, and chronic (prolonged stress). PARP14, a negative regulator of JAK-STAT, had multiple colocalized PTMs that may be involved in intraprotein regulatory crosstalk. Our workflow provides a high-throughput platform that can profile multi-PTMomes from the same sample set, which is valuable in unraveling the functional roles of PTMs and their co-regulation.

Biomass ash was added during pretreatment and enzymatic hydrolysis to examine its influence on biochemical processes for producing fermentable sugars. Corn stover was pretreated using several methods - hot water, dilute acid, alkaline, γ-valerolactone, and ionic liquid methods, each examined with ash loadings of 8.26% and 13.21%. The findings demonstrated that increased ash content adversely affected both pretreatment and enzymatic hydrolysis. Specifically, the total sugar yield was 3–15% lower at the higher ash content across all pretreatment methods, and up to 6.7% lower during enzymatic hydrolysis. For acidic pretreatment, the sugar yield decreased as ash content increased. In contrast, ash content had a lesser impact on alkaline pretreatment compared to acidic pretreatment. The primary reasons for the reduced sugar yield in higher-ash biomass during acidic pretreatments were likely the neutralizing effect of the ash and decreased acid access to the substrates. During enzymatic hydrolysis, ash reduced the sugar yield by limiting enzyme access to cellulose.

Reductive catalytic fractionation (RCF) is a new class of process that produces mono phenolics close-to-theoretical maximum yield by preserving the biomass polysaccharides as solid residues under hydrogen atmo-sphere or hydrogen transfer conditions. Herein, hydrous ruthenium oxide deposited on carbon (HRO@C) and HY-zeolite (HRO@HYZ) catalysts are prepared and characterized by different analytical techniques (TGA, TPD, TPR, XRD, TEM, XPS, physisorption and chemisorption). Both catalysts have been used for the RCF of sugarcane bagasse (SCB). Effects of catalyst acidity, reaction time, pressure, temperature, and solvent on the RCF of SCB have been investigated. Solid residues after RCF are characterized by FT-IR, TGA, XRD, and fibertec to identify the effects of reaction conditions on lignin-first delignification efficiency (LFDE). High acidity of HRO@HYZ favors a high degree of LFDE (81.2 %) than HRO@C (55.8 %). However, alkyl-substituted phenolic monomers (ASPMs) selectivity is 69 % for HRO@C and 55.7 % for HRO@HYZ, respectively, under the same reaction conditions. NMR and GC-FID/GC-MS analysis indicate the presence of ASPMs in the resulted lignin bio-oil. Moreover, fractionated lignin bio-oil is used for upgradation and the upgraded product analysis confirms > 95 % conversion of lignin bio-oil, in which over 90 % selectivity is towards alkylated cyclic alcohols.

Minerals in biomass have a significant impact on both biofuel quality and yield. This is especially true for current thermochemical biomass conversion processes. However, the roles of plant minerals in biochemical conversion have not been studied extensively, even though they are generally considered to lower the sugar yield because they reduce the feedstock proportion of carbohydrates. A successful strategic solution is thus necessary to overcome the challenges caused by the minerals in biomass, which include (1) decreased quality of biomass feedstocks; (2) reduction of process efficiency; and (3) reduction of the product quality and quantity from biomass conversion. This review summarizes the roles of plant minerals in a biorefinery, focusing on these key challenges. The discussion covers many issues related to plant minerals in biofuel production, including their sources, functions, and distribution in plant biomass, methods of characterizing them, their influence in a biorefinery, and the strategic handling required to manage their occurrence in biomass, based on reported studies. It could inspire better strategies to deal with the variance of mineral content in biomass feedstocks to increase process efficiency and reduce costs while supporting the concept of a circular bioeconomy. © 2023 The Authors. Biofuels, Bioproducts and Biorefining published by Society of Industrial Chemistry and John Wiley & Sons Ltd.

Abstract Background Rhodococci are studied for their bacterial ligninolytic capabilities and proclivity to accumulate lipids. Lignin utilization is a resource intensive process requiring a variety of redox active enzymes and cofactors for degradation as well as defense against the resulting toxic byproducts and oxidative conditions. Studying enzyme expression and regulation between carbon sources will help decode the metabolic rewiring that stymies lignin to lipid conversion in these bacteria. Herein, a redox proteomics approach was applied to investigate a fundamental driver of carbon catabolism and lipid anabolism: redox balance. Results A consortium of Rhodococcus strains was employed in this study given its higher capacity for lignin degradation compared to monocultures. This consortium was grown on glucose vs. lignin under nitrogen limitation to study the importance of redox balance as it relates to nutrient availability. A modified bottom–up proteomics workflow was harnessed to acquire a general relationship between protein abundance and protein redox states. Global proteomics results affirm differential expression of enzymes involved in sugar metabolism vs. those involved in lignin degradation and aromatics metabolism. As reported previously, several enzymes in the lipid biosynthetic pathways were downregulated, whereas many involved in β-oxidation were upregulated. Interestingly, proteins involved in oxidative stress response were also upregulated perhaps in response to lignin degradation and aromatics catabolism, which require oxygen and reactive oxygen species and generate toxic byproducts. Enzymes displaying little-to-no change in abundance but differences in redox state were observed in various pathways for carbon utilization (e.g., β‑ketoadipate pathway), lipid metabolism, as well as nitrogen metabolism (e.g., purine scavenging/synthesis), suggesting potential mechanisms of redox-dependent regulation of metabolism. Conclusions Efficient lipid production requires a steady carbon and energy flux while balancing fundamental requirements for enzyme production and cell maintenance. For lignin, we theorize that this balance is difficult to establish due to resource expenditure for enzyme production and stress response. This is supported by significant changes to protein abundances and protein cysteine oxidation in various metabolic pathways and redox processes.

Cyclohexanol and its alkyl derivatives are among the most demanding and valuable chemicals. Currently, cyclohexanols are commercially obtained by oxidation of cyclohexanes, derived from fossil-derived benzene and its derivatives. Herein, cyclohexanols are produced from industrial phenolics (IPs) by hydrodeoxygenation (HDO). HDO of IPs was performed over a series of hydrous ruthenium oxide impregnated HY zeolite (HROY) catalysts with varying Ru content. Highest acidity of 10 HROY promoted deoxygenation reactions and strongly interacted small HRO particles with support led to high hydrogenation ability. The effect of solvent mixture (isopropyl alcohol (IPA): water) demonstrated the positive role of water in improving the conversion and selectivity. IPA facilitated the dissolution of IPs while water promoted the adsorption of solubilized molecules on the HRO active sites to lead the proton transfer activity. Maximum conversion (>= 99%) was achieved with 87.4% cyclohexanols selectivity (2-methyl cyclohexanol:80.4%, 4-methyl cyclohexanol:5.6% and Cyclohexanol: 1.4%) under optimum reaction conditions.

Polylactic acid (PLA) has been used in fused deposition method (FDM) based 3D printing for many years. Alkali lignin is an undervalued industrial by-product that could upgrade PLA's poor mechanical properties. This work presents a biotechnological approach consisting of a partial degradation of alkali lignin using Bacillus ligniniphilus laccase (Lacc) L1 for its use as a nucleating agent in a polylactic acid/thermoplastic polyurethane (PLA/TPU) blend. Results showed that adding enzymatically modified lignin (EL) increased the elasticity modulus to a maximum of 2.5-fold than the control and conferred a maximum biodegradability rate of 15 % after 6 months under the soil burial method. Furthermore, the printing quality rendered satisfactory smooth surfaces, geometries and a tunable addition of a woody color. These findings open a new door for using laccase as a tool to upgrade lignin's properties and its use as a scaffold in manufacturing more environmentally sustainable filaments with improved mechanical properties for 3D printing.