Lignin-based chemicals and biomaterials will be feasible alternatives to their fossil-fuel-based counterparts once their breakdown into constituents is economically viable. The existing commercial market for lignin remains limited due to its complex heterogenous structure and lack of extraction/depolymerization techniques. Hence, in the present study, a novel low-cost ammonium-based protic ionic liquid (PIL), 2-hydroxyethyl ammonium lactate [N11H(2OH)][LAC], is used for the selective fractionation and improved extraction of lignin from Scots pine (Pinus sylvestris) softwood biomass (PWB). The optimization of three process parameters, viz., the incubation time, temperature, and biomass:PIL (BM:PIL) ratio, was performed to determine the best pretreatment conditions for lignin extraction. Under the optimal pretreatment conditions (180 °C, 3 h, and 1:3 BM:PIL ratio), [N11H(2OH)][LAC] yielded 61% delignification with a lignin recovery of 56%; the cellulose content of the recovered pulp was approximately 45%. Further, the biochemical composition of the recovered lignin and pulp was determined and the recovered lignin was characterized using ¹H–¹³C heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) spectroscopy, quantitative ³¹P NMR, gel permeation chromatography (GPC), attenuated total reflectance (ATF)–Fourier transform infrared spectroscopy (ATR-FTIR), and thermal gravimetric analysis (TGA) analysis. Our results reveal that [N11H(2OH)][LAC] is significantly involved in the cleavage of predominant β–O–4’ linkages for the generation of aromatic monomers followed by the in situ depolymerization of PWB lignin. The simultaneous extraction and depolymerization of PWB lignin favors the utilization of recalcitrant pine biomass as feedstock for biorefinery schemes.

Lignin, a vital component of plant biomass, offers significant potential for advanced biorefineries seeking to produce high-value chemicals and materials. However, maximizing lignin yield while ensuring its efficient valorization remains a substantial challenge in biorefineries. In this study, an integrated approach to producing lignin-derived oligomers and monomers was developed from softwood biomass. Pinus sylvestris was processed using a protic ionic liquid (PIL), triethylammonium methane sulfonate ([N222H][OMS]), to extract lignin, followed by its targeted depolymerization using bacterial laccases. Advanced analytical techniques were employed to investigate the qualitative and quantitative changes in lignin during the optimization and depolymerization stages. The findings highlight the effectiveness of [N222H][OMS] in removing 87.90% of lignin from pine wood at 180 °C. Furthermore, engineered bacterial laccases demonstrated significant catalytic activity, converting 9.2% of aliphatic hydroxyl groups and 73.8% of phenolic hydroxyl groups in lignin into carboxylic acids. Similarly, benchmarked against commercially available kraft lignin, the same depolymerization approach achieved lower conversion rates, transforming 12.4% aliphatic and 44.5% phenolic hydroxyl groups in lignin into carboxylic acids. Thus, this integrated strategy, combining ionic liquid delignification with enzymatic upgrading, presents a scalable and efficient route for maximizing lignin valorization.

The valorization of advanced biorefinery lignins remains a significant challenge, owing to the presence of residual carbohydrates. These lignin-associated carbohydrates hinder lignin purification, reduce its homogeneity, and complicate chemical modifications, ultimately limiting the efficient conversion of lignin into high-value products such as chemicals and materials. This study presents a protic ionic liquid-based lignin fractionation process developed using softwood biomass. Triethylammonium methane sulfonate ([N222H][OMS]) was used to fractionate Pinus sylvestris, yielding two distinct fractions: a low-molecular-weight lignin fraction (LF) and a high-molecular-weight lignin fraction (HF). The extracted fractions were comprehensively characterized using nuclear magnetic resonance (NMR) to quantify changes in interunit linkages (β-O-4, β-5, and β-β) and hydroxyl group distribution, whereas methanolysis gas chromatography/mass spectrometry (GC/MS) was used to quantify residual carbohydrates. The fractionation process achieved LF and HF yields of approximately 70.32% and 17.58%, respectively. Further analysis revealed that the HF contained 59.92 ± 2.12 mg/g carbohydrates, whereas the LF contained only 27.37 ± 1.13 mg/g. These findings underscore the effectiveness of the protic ionic liquid fractionation process in reducing carbohydrate impurities and enhancing lignin purity, paving the way for the more efficient utilization of lignin in value-added applications.

Lignin is an abundant and renewable source capable of replacing different raw materials in the chemical industry. It can be obtained from lignocellulosic biomass (LCB) via different pretreatment methods. In the present study, hydrolysis lignin (HL) from the Sunburst^TM pretreatment technology was utilized to investigate its enzymatic conversion. At first, soluble HL fractions were obtained via alkali solubilization followed by acid precipitation, referred to as acid precipitated lignin (APL). Furthermore, the APL was tested with three different bacterial laccases to identify the optimal conditions for its conversion into small molecular weight fractions. Among the tested laccases, Streptomyces coelicolor A3(2) (ScLac) displayed the highest rate of APL conversion with a high lignin dosage and under extremely alkaline conditions, i.e., 50 g/L in 0.25 M NaOH solution, resulting in higher molecular weight fractions. The increase in the molecular weight and quantitative linkages before and after the enzymatic oxidation of the APL were characterized by size exclusion chromatography (SEC), Fourier-transform infrared spectroscopy (FT-IR), and two-dimensional heteronuclear single quantum correlation nuclear magnetic resonance (2D HSQC NMR) methods.

An integrated lignin-focused biorefining approach.

The transition to a forest-based economy relies on sustainable alternatives that can efficiently convert renewable resources into chemical and material pro