Guaiacylglycerol-β-guaiacyl Ether Research Articles

Fungal enzyme degradation of lignin-PLA composites: Insights from experiments and molecular docking simulations

Fungal enzymes are effective in degrading various polymeric materials. In this study, we assessed the initial degradation of composites consisting of lignin-poly(lactic acid) (PLA) with both unmodified lignin (LIG) and oxypropylated lignin (oLIG) incorporated at 10 % and 40 % weight within the PLA matrix in a fungal environment. Trametes versicolor fungi were used, and the samples were treated only for eight weeks. Although there was no significant difference in weight loss, the degradation process impacted the chemical and thermal properties of the composites, as shown by Fourier transform infrared spectroscopy (FTIR) and Differential scanning calorimetry (DSC) analyses. After the degradation process, the carbonyl index values decreased for all composites and the hydroxyl index values increased for LIG/PLA and a reverse trend was observed for oLIG/PLA composites. The first heating scan from DSC results showed that the melting peak and the cold crystallization peak disappeared after the degradation process. Microscopic analysis revealed that LIG/PLA exhibited higher roughness than oLIG/PLA. Molecular docking simulations were carried out using guaiacylglycerol-β-guaiacyl ether (GGE) and lactic acid (LA) as model compounds for lignin and PLA, respectively, with laccase (Lac) enzyme for Trametes versicolor. The docking results showed that GGE had the strongest binding interaction and affinity with Lac than lactic acid and oxypropylated GGE. The oxypropylated GGE formed a shorter hydrogen bonding with the Lac enzyme than GGE and LA. The trend associated with the degradation of composites from experimental and molecular docking findings was consistent. This combined approach provided insights into the degradation process using fungi and had the potential to be applied to different polymeric composites.

Design and Engineering of an Efficient Peroxidase Using Myoglobin for Dye Decolorization and Lignin Bioconversion.

The treatment of environmental pollutants such as synthetic dyes and lignin has received much attention, especially for biotechnological treatments using both native and artificial metalloenzymes. In this study, we designed and engineered an efficient peroxidase using the O2 carrier myoglobin (Mb) as a protein scaffold by four mutations (F43Y/T67R/P88W/F138W), which combines the key structural features of natural peroxidases such as the presence of a conserved His-Arg pair and Tyr/Trp residues close to the heme active center. Kinetic studies revealed that the quadruple mutant exhibits considerably enhanced peroxidase activity, with the catalytic efficiency (kcat/Km) comparable to that of the most efficient natural enzyme, horseradish peroxidase (HRP). Moreover, the designed enzyme can effectively decolorize a variety of synthetic organic dyes and catalyze the bioconversion of lignin, such as Kraft lignin and a model compound, guaiacylglycerol-β-guaiacyl ether (GGE). As analyzed by HPLC and ESI-MS, we identified several bioconversion products of GGE, as produced via bond cleavage followed by dimerization or trimerization, which illustrates the mechanism for lignin bioconversion. This study indicates that the designed enzyme could be exploited for the decolorization of textile wastewater contaminated with various dyes, as well as for the bioconversion of lignin to produce more value-added products.

A Novel Polyphenol Oxidoreductase OhLac from Ochrobactrum sp. J10 for Lignin Degradation.

Identifying the enzymes involved in lignin degradation by bacteria is important in studying lignin valorization to produce renewable chemical products. In this paper, the catalytic oxidation of lignin by a novel multi-copper polyphenol oxidoreductase (OhLac) from the lignin degrader Ochrobactrum sp. J10 was explored. Following its expression, reconstitution, and purification, a recombinant enzyme OhLac was obtained. The OhLac enzyme was characterized kinetically against a range of substrates, including ABTS, guaiacol, and 2,6-DMP. Moreover, the effects of pH, temperature, and Cu2+ on OhLac activity and stability were determined. Gas chromatography-mass spectrometer (GC-MS) results indicated that the β-aryl ether lignin model compound guaiacylglycerol-β-guaiacyl ether (GGE) was oxidized by OhLac to generate guaiacol and vanillic acid. Molecular docking analysis of GGE and OhLac was then used to examine the significant amino residues and hydrogen bonding sites in the substrate–enzyme interaction. Altogether, we were able to investigate the mechanisms involved in lignin degradation. The breakdown of the lignocellulose materials wheat straw, corn stalk, and switchgrass by the recombinant OhLac was observed over 3 days, and the degradation results revealed that OhLac plays a key role in lignin degradation.

Biotransformation of Lignin by an Artificial Heme Enzyme Designed in Myoglobin With a Covalently Linked Heme Group.

The conversion of Kraft lignin in plant biomass into renewable chemicals, aiming at harvesting aromatic compounds, is a challenge process in biorefinery. Comparing to the traditional chemical methods, enzymatic catalysis provides a gentle way for the degradation of lignin. Alternative to natural enzymes, artificial enzymes have been received much attention for potential applications. We herein achieved the biodegradation of Kraft lignin using an artificial peroxidase rationally designed in myoglobin (Mb), F43Y/T67R Mb, with a covalently linked heme cofactor. The artificial enzyme of F43Y/T67R Mb has improved catalytic efficiencies at mild acidic pH for phenolic and aromatic amine substrates, including Kraft lignin and the model lignin dimer guaiacylglycerol-β-guaiacyl ether (GGE). We proposed a possible catalytic mechanism for the biotransformation of lignin catalyzed by the enzyme, based on the results of kinetic UV-Vis studies and UPLC-ESI-MS analysis, as well as molecular modeling studies. With the advantages of F43Y/T67R Mb, such as the high-yield by overexpression in E. coli cells and the enhanced protein stability, this study suggests that the artificial enzyme has potential applications in the biodegradation of lignin to provide sustainable bioresource.

Bioconversion of Guaiacylglycerol-β-guaiacyl ether into vanillin by Burkholderia sp. ISTR5: A genomic and proteomic approach

Long-term research on biofuel production from lignocellulose did not progress much due to strong inter-unit linkages in lignin structure. Lignin contains β-O-4 linkage as a major bond (50–60%) between aromatic compounds to form a complex 3D-structure. The present study aims for the characterization of a novel strain Burkholderia sp. ISTR5 (R5) for β-O-4 linkage disintegration. Guaiacylglycerol-β-guaiacyl ether (GGE) was taken as a representative compound for β-O-4 linkage analysis. We showed that the strain R5 completely degraded GGE in 72 h. Further, gas chromatography-mass spectrometry (GC-MS) profile showed that vanillin is one of the highest produced chemicals during the degradation of GGE with small fractions of other metabolites in the reaction mixture. Nuclear Magnetic Resonance (NMR) analysis demonstrated the fragmentation of β-O-4 bonds and the advent of new chemical bonds. In addition to it, the whole genome analysis showed that R5 contains a superior enzyme chassis for the degradation of aromatic compounds. Enzymes such as glutathione-s-transferase, biphenyl dioxygenase, catalase/peroxidase, hydrolases, and dehydrogenases dominated the genome of R5. Proteomics analysis showed 2 glutathione lyase, 1 glutathione-s-transferase, 1 GMC oxidoreductase, 2 FAD binding oxidoreductase and several hydrolases, peroxidases, and transferases were upregulated on GGE. This study showed the capability of R5 to degrade and bioconvert the β-O-4 linkage lignin model compound GGE into vanillin.

Revealing of Supercritical Water Gasification Process of Lignin by Reactive Force Field Molecular Dynamics Simulations

Gasification with supercritical water is an efficient process that can be used for the valorization of biomass. Lignin is the second most abundant biopolymer in biomass and its conversion is fundamental for future energy and value-added chemicals. In this paper, the supercritical water gasification process of lignin by employing reactive force field molecular dynamics simulations (ReaxFF MD) was investigated. Guaiacyl glycerol-β-guaiacyl ether (GGE) was considered as a lignin model to evaluate the reaction mechanism and identify the components at different temperatures from 1000 K to 5000 K. The obtained results revealed that the reactions and breaking of the lignin model started at 2000 K. At the primary stage of the reaction at 2000 K the β-O-4 bond tends to break into several compounds, forming mainly guaiacol and 1,3-benzodioxole. In particular, 1,3-benzodioxole undergoes dissociation and forms cyclopentene-based ketones. Afterward, dealkylation reaction occurred through hydroxyl radicals of water to form methanol, formaldehyde and methane. Above 2500 K, H2, CO and CO2 are predominantly formed in which water molecules contributed hydrogen and oxygen for their formation. Understanding the detailed reactive mechanism of lignin’s gasification is important for efficient energy conversion of biomass.

Degradation of a Lignin Model Compound by ROS Generated In Situ through Controlled ORR in Ionic Liquid

The cleavage of the linkage bonds among the C9 units in lignin molecules via selective oxidation method to obtain small-weight aromatic compounds is an important way to lignin valorisation. In this study, the cleavage pathways of bonds in a lignin model compound, GGE (guaiacylglycerol-β-guaiacyl ether), by the reactive oxygen species (ROS) in situ generated through electrochemical oxygen reduction reaction (ORR) in an aprotic ionic liquid ([BMIM]BF4) was investigated. The results indicated that when the main ROS was ·OOH, coniferyl alcohol, guaiacol and vanillin were detected as the degradation products which is supposed to be caused by β-O-4 bonds cleavage; Otherwise, when HO2 − was the main ROS, coniferyl alcohol could not be detected and the degradation reaction is believed to be trigerred by C α -C β bonds cleavage.

A two-phase reaction system for selective oxidative degradation of lignin model compounds

Lignin depolymerization through an oxidation method could provide value-added products, but it is challenging in terms of recovering catalysts or separating products in time to avoid over-oxidation. In this study, a process of selectively oxidative degradation of lignin model compounds was operated in a two-phase reaction system. Lignin model compounds of 4-benzyloxyphenol (PBP) or guaiacylglycerol-β-guaiacyl ether (GGE) in a bottom phase of 1-butyl-3-methylimidazole chloride ([BMIM]Cl) ionic liquid were selectively oxidized by H2O2 in the presence of a solid acid (SO42-/Fe2O3-ZrO2), and the degradation products immediately diffused into the upper organic solvent phase (butyl acetate). In this kind of reaction system, the yield of the products was improved due to the prolonged life of ∙OH in ionic liquid, and the product selectivity was maintained due to the timely product separation, and the ionic liquid and the catalyst were easily recycled.

Ru-Catalyzed Oxidative Cleavage of Guaiacyl Glycerol-β-Guaiacyl Ether-a Representative β-O-4 Lignin Model Compound

The introduction of efficient and selective catalytic methods for aerobic oxidation of lignin and lignin model compounds to aromatics can extend the role of lignin applications in biorefineries. The current study focussed on the catalytic oxidative transformation of guaiacyl glycerol-β-guaiacyl ether (GGGE)–a β-O-4 lignin model compound to produce basic aromatic compounds (guaiacol, vanillin and vanillic acid) using metal-supported catalysts. Ru/Al2O3, prepared with ruthenium(IV) oxide hydrate, showed the highest yields of the desired products (~60%) in acetonitrile in a batch reactor at 160 °C and 5-bar of 20% oxygen in argon. Alternative catalysts containing other transition metals (Ag, Fe, Mn, Co and Cu) supported on alumina, and ruthenium catalysts based on alternative supports (silica, spinel, HY zeolite and zirconia) gave significantly lower activities compared to Ru/Al2O3 at identical reaction conditions. Moreover, the Ru/Al2O3 catalyst was successfully reused in five consecutive reaction runs with only a minor decrease in catalytic performance.

Degradation of the phenolic β-ether lignin model dimer and dyes by dye-decolorizing peroxidase from Bacillus amyloliquefaciens.

The dye-decolorizing peroxidase from Bacillus amyloliquefaciens, BaDyP, was identified to be an efficient catalyst for the degradation of phenolic β-ether lignin model dimer guaiacylglycerol-β-guaiacyl ether (GGE) and dyes. Efeb gene encoding BaDyP from B. amyloliquefaciens MN-13 consisted of 1257 bp and the open reading frame encoded 418 amino acids. The efeb gene was expressed in Escherichia coli BL21 and a recombinant BaDyP of 50 kDa was achieved. The BaDyP exhibited activity in oxidizing GGE and decolorizing azo and triphenylmethane dyes. At pH 4.5 and 30°C the BaDyP not only completely degraded GGE by the cleavage of β-O-4 ether bond and Cα-Cβ bond, and Cα oxidation without any oxidative mediator, but also decolorized four synthetic dyes, including congo red, bromine cresol green, eriochrome black T and crystal violet. This was achieved with decolorization rates of 65.7%, 70.62%, 80.06% and 62.09%, respectively, after 72 h of incubation. BaDyP was identified as a bacteria peroxidase with great potential for the degradation of lignin and bioremediation of dye-contamination.

Theoretical Elucidation of β-O-4 Bond Cleavage of Lignin Model Compound Promoted by Sulfonic Acid-Functionalized Ionic Liquid.

While the depolymerization of lignin to chemicals catalyzed by ionic liquids has attracted significant attention, the relevant molecular mechanism, especially the cleavage of specific bonds related to efficient depolymerization, still needs to be deeply understood for the complexity of this natural aromatic polymer. This work presents a detailed understanding of the cleavage of the most abundant β-O-4 bond in the model system, guaiacylglycerol β-guaiacyl ether, by a Brønsted acidic IL (1-methyl-3-(propyl-3-sulfonate) imidazolium bisulfate ([C3SO3Hmim][HSO4]) using density functional theory calculation and molecular dynamics simulation. It has been found that [C3SO3Hmim][HSO4] generates zwitterion/H2SO4 via proton transfer with an energy barrier of 0.38 kcal/mol, which plays a dominant role in the lignin depolymerization process. Subsequently, the reaction can be carried out via three potential pathways, including (1) the dehydration of α-C-OH, (2) dehydration of γ-C-OH, and (3) the protonation of β-O. The electrophilic attack of H2SO4 and the hydrogen-bonding interaction between GG and zwitterion are the two most important factors to promote the depolymerization reaction. In all steps, the dehydration of α-C-OH route is computed to be favored for the experiment. The relatively higher energy barrier for β-O-4 bond dissociation among these reaction steps is attributed to the hindrance of the self-assembled clusters of GG in the mixed system. Further, the dense distribution of H13([C3SO3Hmim]) surrounding O21(GG), indicated by sharp peaks in RDFs, reveals that -SO3H in cations plays a substantial role in solvating lignin. Hopefully, this work will demonstrate new insights into lignin depolymerization by functionalized ILs in biomass conversion chemistry.

Characterization and use of a bacterial lignin peroxidase with an improved manganese-oxidative activity.

Peroxidases are well-known biocatalysts produced by all organisms, especially microorganisms, and used in a number of biotechnological applications. The enzyme DypB from the lignin-degrading bacterium Rhodococcus jostii was recently shown to degrade solvent-obtained fractions of a Kraft lignin. In order to promote the practical use, the N246A variant of DypB, named Rh_DypB, was overexpressed in E. coli using a designed synthetic gene: by employing optimized conditions, the enzyme was fully produced as folded holoenzyme, thus avoiding the need for a further time-consuming and expensive reconstitution step. By a single chromatographic purification step, > 100mg enzyme/L fermentation broth with a > 90% purity was produced. Rh_DypB shows a classical peroxidase activity which is significantly increased by adding Mn2+ ions: kinetic parameters for H2O2, Mn2+, ABTS, and 2,6-DMP were determined. The recombinant enzyme shows a good thermostability (melting temperature of 63-65°C), is stable at pH6-7, and maintains a large part of the starting activity following incubation for 24h at 25-37°C. Rh_DypB activity is not affected by 1M NaCl, 10% DMSO, and 5% Tween-80, i.e., compounds used for dye decolorization or lignin-solubilization processes. The enzyme shows broad dye-decolorization activity, especially in the presence of Mn2+, oxidizes various aromatic monomers from lignin, and cleaves the guaiacylglycerol-β-guaiacyl ether (GGE), i.e., the Cα-Cβ bond of the dimeric lignin model molecule of β-O-4 linkages. Under optimized conditions, 2mM GGE was fully cleaved by recombinant Rh_DypB, generating guaiacol in only 10min, at a rate of 12.5μmol/minmg enzyme.

Multienzyme immobilized polymeric membrane reactor for transformation of lignin model compound.

We have developed an integrated, multienzyme functionalized membrane reactor for bioconversion of a lignin model compound involving enzymatic catalysis. The membrane bioreactors were fabricated through the layer-by-layer assembly approach to immobilize three different enzymes (glucose oxidase, peroxidase and laccase) into pH-responsive membranes. This novel membrane reactor couples the in situ generation of hydrogen peroxide (by glucose oxidase) to oxidative conversion of a lignin model compound, guaiacylglycerol-β-guaiacyl ether (GGE). Preliminary investigation of the efficacy of these functional membranes towards GGE degradation is demonstrated under convective flow mode. Over 90% of the initial feed could be degraded with the multienzyme immobilized membranes at a residence time of approximately 22 s. GGE conversion product analysis revealed the formation of oligomeric oxidation products upon reaction with peroxidase, which may be a potential hazard to membrane bioreactors. These oxidation products could further be degraded by laccase enzymes in the multienzymatic membranes, explaining the potential of multi enzyme membrane reactors. The multienzyme incorporated membrane reactors were active for more than 30 days of storage time at 4 °C. During this time span, repetitive use of the membrane reactor was demonstrated involving 5–6 h of operation time for each cycle. The membrane reactor displayed encouraging performance, losing only 12% of its initial activity after multiple cycles of operation.

Novosphingobium aromaticivorans uses a Nu-class glutathione S-transferase as a glutathione lyase in breaking the β-aryl ether bond of lignin

As a major component of plant cell walls, lignin is a potential renewable source of valuable chemicals. Several sphingomonad bacteria have been identified that can break the β-aryl ether bond connecting most phenylpropanoid units of the lignin heteropolymer. Here, we tested three sphingomonads predicted to be capable of breaking the β-aryl ether bond of the dimeric aromatic compound guaiacylglycerol-β-guaiacyl ether (GGE) and found that Novosphingobium aromaticivorans metabolizes GGE at one of the fastest rates thus far reported. After the ether bond of racemic GGE is broken by replacement with a thioether bond involving glutathione, the glutathione moiety must be removed from the resulting two stereoisomers of the phenylpropanoid conjugate β-glutathionyl-γ-hydroxypropiovanillone (GS-HPV). We found that the Nu-class glutathione S-transferase NaGSTNu is the only enzyme needed to remove glutathione from both (R)- and (S)-GS-HPV in N. aromaticivorans. We solved the crystal structure of NaGSTNu and used molecular modeling to propose a mechanism for the glutathione lyase (deglutathionylation) reaction in which an enzyme-stabilized glutathione thiolate attacks the thioether bond of GS-HPV, and the reaction proceeds through an enzyme-stabilized enolate intermediate. Three residues implicated in the proposed mechanism (Thr51, Tyr166, and Tyr224) were found to be critical for the lyase reaction. We also found that Nu-class GSTs from Sphingobium sp. SYK-6 (which can also break the β-aryl ether bond) and Escherichia coli (which cannot break the β-aryl ether bond) can also cleave (R)- and (S)-GS-HPV, suggesting that glutathione lyase activity may be common throughout this widespread but largely uncharacterized class of glutathione S-transferases.

Perspectives for biocatalytic lignin utilization: cleaving 4-O-5 and C\u03b1\u2013C\u03b2 bonds in dimeric lignin model compounds catalyzed by a promiscuous activity of tyrosinase

BackgroundIn the biorefinery utilizing lignocellulosic biomasses, lignin decomposition to value-added phenolic derivatives is a key issue, and recently biocatalytic delignification is emerging owing to its superior selectivity, low energy consumption, and unparalleled sustainability. However, besides heme-containing peroxidases and laccases, information about lignolytic biocatalysts is still limited till date.ResultsHerein, we report a promiscuous activity of tyrosinase which is closely associated with delignification requiring high redox potentials (>1.4 V vs. normal hydrogen electrode [NHE]). The promiscuous activity of tyrosinase not only oxidizes veratryl alcohol, a commonly used nonphenolic substrate for assaying ligninolytic activity, to veratraldehyde but also cleaves the 4-O-5 and Cα–Cβ bonds in 4-phenoxyphenol and guaiacyl glycerol-β-guaiacyl ether (GGE) that are dimeric lignin model compounds. Cyclic voltammograms additionally verified that the promiscuous activity oxidizes lignin-related high redox potential substrates.ConclusionThese results might be applicable for extending the versatility of tyrosinase toward biocatalytic delignification as well as suggesting a new perspective for sustainable lignin utilization. Furthermore, the results provide insight for exploring the previously unknown promiscuous activities of biocatalysts much more diverse than ever thought before, thereby innovatively expanding the applicable area of biocatalysis.

Investigation of lignin biodegradation by Trabulsiella sp. isolated from termite gut

Lignocellulosic biomass is a promising source of several value-added chemicals besides being used to produce pyrolytic oils. Microbial degradation of lignin is an effective method of depolymerizing it into smaller organic chemicals. The aim of the present study is to screen termite gut microbial flora for their ability to degrade alkyl lignins. Experiments have been carried out initially with guaiacylglycerol-β-guaiacyl ether (GGE), a lignin model compound. Microbes isolated from Odontotermes obesus gut were screened for GGE degradation efficiency. Out of several strains, the IIPTG13 which is closely related to the Trabulsiella sp. bacterium, showed maximum GGE degradation. Hence, it was eventually tested for alkyl lignin degradation at different pH and temperatures. Residual GGE and degraded products of alkyl lignin were analyzed using gas chromatography coupled to a mass spectrometry (GC-MS). Microbial degradation of alkyl lignin showed several organic compounds in GC-MS analysis which indicates that IIPTG13 strain has the potential to degrade lignin.

Phylogenetic and kinetic characterization of a suite of dehydrogenases from a newly isolated bacterium, strain SG61-1L, that catalyze the turnover of guaiacylglycerol-β-guaiacyl ether stereoisomers.

Lignin is a complex aromatic polymer found in plant cell walls that makes up 15 to 40% of plant biomass. The degradation of lignin substructures by bacteria is of emerging interest because it could provide renewable alternative feedstocks and intermediates for chemical manufacturing industries. We have isolated a bacterium, strain SG61-1L, that rapidly degrades all of the stereoisomers of one lignin substructure, guaiacylglycerol-β-guaiacyl ether (GGE), which contains a key β-O-4 linkage found in most intermonomer linkages in lignin. In an effort to understand the rapid degradation of GGE by this bacterium, we heterologously expressed and kinetically characterized a suite of dehydrogenase candidates for the first known step of GGE degradation. We identified a clade of active GGE dehydrogenases and also several other dehydrogenases outside this clade that were all able to oxidize GGE. Several candidates exhibited stereoselectivity toward the GGE stereoisomers, while others had higher levels of catalytic performance than previously described GGE dehydrogenases for all four stereoisomers, indicating a variety of potential applications for these enzymes in the manufacture of lignin-derived commodities.

Convenient synthesis of chiral lignin model compounds via optical resolution: four stereoisomers of guaiacylglycerol-β-guaiacyl ether and both enantiomers of 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-propan-1-one (erone)

Guaiacylglycerol-β-guaiacyl ether (GGE) is one of the most important phenolic compound for studying the chemistry and biochemistry of lignin. GGE contains two asymmetric carbons at the alpha and beta positions of its side chain; therefore, theoretically it can exist as four different stereoisomers. It has been proposed that a Gram-negative bacterium, Sphingobium sp. SYK-6 (formerly referred as Sphingomonas paucimobilis SYK-6), degrades GGE enantiospecifically via cleavage of the ether bond. The cleavage was thought to proceed in two steps, each catalyzed by a different enantiospecific enzyme. In the first step, the alcohol residue at the alpha position of the side chain in GGE was thought to be oxidized enantiospecifically by four distinct Cα-dehydrogenases (LigD, LigN, LigL and LigO), to produce two enantiomers of 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-propan-1-one (erone). To study this enantiospecific degradation step by the four dehydrogenases, we synthesized all four stereoisomers of GGE and both enantiomers of erone and separated them into enantiopure samples. The stereoisomers and our synthetic methods to prepare them are useful both for microbial and chemical investigations of lignin degradation.

Reactivity of long chain alkylamines to lignin moieties: Implications on hydrophobicity of lignocellulose materials

Enzymatic processes provide new perspectives for modification of lignocellulose materials. In the current study, laccase catalyzed coupling of long chain alkylamines to lignin model molecules and lignocellulose was investigated. Up to two molecules of dodecylamine (DA) and dihexylamine (DHA) were successfully coupled with lignin monomers (guaiacol, catechol and ferulic acid) while coupling onto complex lignin model compounds (syringylglycerol β-guaiacyl ether, guaiacylglycerol β-guaiacyl ether and dibenzodioxocin) yielded 1:1 coupling products. Surface analysis of beech veneers enzymatically grafted with DA showed an increase in nitrogen content of 3.18% compared to 0.71% in laccase only treated controls while the O/C ratio decreased from 0.52 to 0.46. Concomitantly the grafting of DHA or DA onto beech veneers resulted in a 53.8% and 84.2% increase in hydrophobicity, respectively when compared to simple adsorption. Therefore, laccase-mediated grafting of long chain alkylamines onto lignocellulose materials can be potentially exploited for improving their hydrophobicity.

The closed system pyrolysis of β-O-4 lignin substructure model compounds

Lignin substructure model compounds having a β-O-4 linkage were synthesized. These were guaiacylglycerol-β-guaiacyl ether (GGE) and guaiacylglycerol-β-syringyl ether (GSE) which model guaiacyl and syringyl units respectively. Closed system microscale pyrolysis of GGE and GSE was carried out at 300 °C both in the presence and absence of water vapour. The laboratory degradation of GSE occurred predominantly by breaking the Cβ–O bond of the β-O-4 linkage to form the ring B fragment of the model compound. This was followed by demethylation of the aromatic methoxyl groups. Char formation was another significant process during pyrolysis. There are structural features of the char formed during the heating of GGE which indicate incorporation of the remaining fragment of the model compound (ring A coupled with the propyl segment of the aryl ether linkage).