The comparison of chemical and structural differences of Caribean pine (Pinus caribea L.) wood and bark lignin

The presented paper deals with some chemical properties of Caribean pine wood and bark lignin. In order to obtain necessary data, the following operations and analyses were performed: dioxane lignin isolations, determination of Klason lignin, determination of methoxyl, carboxyl and phenolic hydroxyl groups in dioxane lignins and the nitrobenzene oxidation of both compared lignins and of the risidues remaining after dioxane lignins extraction. The oxidation products were analysed by GLC. The experimental data show that there are moderate differences between compared lignins.     

14C labelling of lignins of normal and bm3 maizes for fermentation studies

[U-¹⁴C] phenylalanine (phe*) and [O¹⁴CH₃] sinapic acid (sin*) were infused into the cut ends of normal and bm3 maizes (anthesis stage) under or above the last node or at mid-internode, with or without the leaf, in light or in darkness. Radioactivity was measured in the organs, and in phenolic constituents of the cell wall and saponified residues of the bases and tops of the apical inter-node. In both maize genotype labelled under the node the radioactivity was distributed more evenly in the organs with sin* than with phe*. Infusion above the node and at mid-internode greatly increased radioactivity in the bases and tops, respectively. Removal of the leaf only slightly increased the radioactivity, mainly in the bases, and no clear-cut effect of darkness was observed. Phe* labelled the phenolic acids and the three lignin units, but the syringyl units of bm3 maize were only slightly labelled. Sin* specifically labelled the syringyl units, which represented the least condensed fraction of lignins. Both the native and labelled lignins were highly alkali soluble. There were differences in lignin biogenesis between the bases and tops, and between normal and bm3 maizes. The newly formed lignins were slightly different from the native lignins but had similar types of heterogeneity, with variations in the internode and between genotypes similar to those in native lignins. Provided due allowance is made for the distinguishing characteristics of newly formed lignins, the [¹⁴C-lignin] cell walls, which are strongly labelled on complementary structures, seem suitable model substrates for fermentation studies.     

Changes on Content,Structure and Surface Distribution of Lignin in Jute Fibers After Laccase Treatment

Effect of laccase treatment on the content, structure, and surface distribution of lignin in jute fibers were fundamentally investigated. Four percent lignin was removed from jute fibers via the laccase treatment. The residual lignin in the laccase-treated jute fibers showed increased molecular weights, which indicated polymerization between lignins on jute fibers. Meanwhile, the phenolic hydroxyl content in lignin decreased during the laccase oxidation accompanied by demethylation of methoxyl groups and generation of carbonyl groups. Due to the degradation and subsequent polymerization of lignin by laccase, the bulgy lignins on jute fiber surfaces were redistributed, which made the surface neat and glossy.     

Comparative study of Caribbean pine (Pinus caribaea L.) wood and bark dioxane lignin

The objective of this paper was to depict some differences between Caribbean pine bark and wood lignin. Gas chromotography (GC) of nitrobenzene oxidation products of extracted wood and bark meals and of corresponding dioxane lignins was performed. The isolated lignins were submitted to IR and UV spectrometry, gel permeation chromatography and differential thermo-gravimetry. The data obtained indicate differences in the contents of condensed bis-guaiacyl structures in the lignins. Contents of phenolic hydroxyl and α-carbonyl groups in both lignins do not markedly differ. There are also certain differences in the thermostability and the molecular weights of compared dioxane lignin preparations.     

Biological variability in lignification of maize: Expression of the brown midrib bm2 mutation

The cell wall phenolic components in the internodes of three maize genotypes, namely normal, bm2 and bm3 maize, were determined. The bm2 and bm3 brown midrib mutations lowered the lignin content of the bottom, middle and top internodes to a similar extent. However, unlike bm3, the bm2 trait did not induce a sharp reduction of the level of ester-bound p-coumaric acid in maize internodes. The other main alkali labile phenolic acid, ferulic acid, reached similar levels in the three genotypes. The main difference between bm2 and bm3 mutations occurred in the alkyl aryl ether linked structures of the lignin component. In contrast to bm3 lignins, which are characterised by a low syringyl content, the bm2 lignin had a lower content of guaiacyl units than lignin of normal maize internode. Consequently, the syringyl/guaiacyl molar ratio of bm2 lignin gave higher values (2.7–3.2) than those from either normal (0.9–1.5) or bm3 lignins (0.3). The alkali solubility of lignin was also compared between the three genotypes. Incorporation of the bm3 trait in maize led to a high recovery of alkali soluble lignin whereas the bm2 lignin had a similar solubility to the normal one in 2 M NaOH. The monomeric composition of the alkali soluble lignins was consistent with the non-condensed structures of the in-situ polymer. Although the bm3 and bm2 mutations had different effects on lignification, the modification of the cell wall phenolic level was also found in the bm2 maize stem as previously studied.     

Comparative characterisation of dioxane-soluble lignins released by ball milling and by sheep digestion from forage grasses

Faecal soluble lignin (FSL), extracted from the faeces of sheep that received orchardgrass and timothy, were compared with 90% dioxane-soluble lignins released by ball milling (MHL) and by subsequent enzymatic hydrolysis (EHL) from the same grasses. FSL contained much less carbohydrate and esterified p-coumaric and ferulic acids than MHL and EHL. Although no considerable differences were found in the yields of vanillin and syringaldehyde produced by nitrobenzene oxidation, the molar ratio of syringaldehyde to vanillin was higher for FSL than for MHL and EHL. Such differences in chemical properties among the lignin preparations were confirmed by ¹³C-NMR spectroscopy. FSL had a lower molecular size compared with MHL and EHL. Results indicated that syringyl-rich lignin fragments with less phenolic acid esters, probably depolymerisation products of the grass lignins, were released by digestion in sheep from the forage grasses.     

Microwave-assisted organic acid extraction of lignin from bamboo: Structure and antioxidant activity investigation

Microwave-assisted extraction in organic acid aqueous solution (formic acid/acetic acid/water, 3/5/2, v/v/v) was applied to isolate lignin from bamboo. Additionally, the structural features of the extracted lignins were thoroughly investigated in terms of C₉ formula, molecular weight distribution, FT-IR, ¹H NMR and HSQC spectroscopy. It was found that with an increase in the severity of microwave-assisted extraction, there was an increase of phenolic hydroxyl content in the lignin. In addition, an increase of the severity resulted in a decrease of the bound carbohydrate content as well as molecular weight of the lignin. Antioxidant activity investigation indicated that the radical scavenging index of the extracted lignins (0.35–1.15) was higher than that of BHT (0.29) but lower than that of BHA (3.85). The results suggested that microwave-assisted organic acid extraction provides a promising way to prepare lignin from bamboo with good antioxidant activity for potential application in the food industry.     

Transformations of (14C-lignin) cell walls of wheat by rumen microorganisms

The lower halves of apical internodes of wheat harvested at the flowering stage were labelled with [U-¹⁴C] phenylalanine (phe) or with [O¹⁴CH₃] sinapic acid (sin). Cell wall residues (CWR) and saponified residues (SR) were incubated in a fermenter simulating the rumen for 7 days with rumen fluid or without microorganisms (controls). PheCWR was labelled in all lignin units (measured as aldehydes from nitrobenzene oxidation), in phenolic acids and slightly in proteins. Labelling of pheSR was more lignin-specific. SinCWR and sinSR were specifically labelled in syringyl units of lignin. The fermentation of CWR resulted in phenylpropane-derived unit losses in the following decreasing order: ferulic acid>p-coumaric acid>syringaldehyde>vanillin>p-hydroxybenzaldehyde. If allowance is made for slight losses in controls, 61, 52, 61 and 63% of the phenylpropanes of pheCWR, sinCWR, pheSR and sinSR, respectively, were transformed into an acid-precipitable fraction, an acid-soluble fraction and ¹⁴CO_2. The comparison of pheCWR and sinCWR degradation showed that syringyl units were solubilised into acid-precipitable molecules to a greater extent than the other lignin units; demethylation of the syringyl units of lignins was also evident from the different productions of ¹⁴CO₂. Alkali-resistant lignins of SR were mainly transformed into acid-precipitable molecules and were weakly degraded. Lignin solubilisation and degradation seem to be governed by different mechanisms which depend on both cell wall structure and rumen microflora.     

Determination of lignin in herbaceous plants by an improved acetyl bromide procedure

A modified acetyl bromide procedure is proposed for the spectrophotometric determination of lignin in herbaceous plants. The digestion with 25 % acetyl bromide (AcBr) in acetic acid at 70°C is improved by including 4 % perchloric acid in the digesting solutions. This allows for more rapid, complete digestion and the use of coarser samples. On the basis of infrared spectra and nitrobenzene oxidation data for milled sample lignins, the value of 20.0 g^?1 litre cm^?1 for the specific absorption coeficient of AcBr-treated lignins of herbaceous plant samples gave lignin values which were consistent with nitrobenzene oxidation data, but were higher than those obtained by the acid detergent lignin method and the earlier AcBr method proposed for grasses. The lower lignin values obtained in the latter methods are considered to be due to partial lignin solubilisation in the sulphuric acid digestion.     

Chemical and in vivo evaluation of a brown midrib mutant of Zea mays. II. Nuclear magnetic resonance spectra of digested and undigested alkali lignins and undigested dimethyformamide lignins

Nuclear magnetic resonance (n.m.r.) spectra were obtained on the alkali lignins extracted from Tr and bm₁ cornstalks, before and after digestion by sheep, and on the dimethylformamide lignins (DMFL) from undigested cornstalks. Differences due to solvent extraction, plant genotype and digestion were apparent. The bm¹ corn lignin has a higher degree of cross linking of the propane side chain of the lignin molecule than Tr corn lignin, a factor which may be significant in determining the effect of lignin on digestibility. The main effect of digestion was to increase the degree of cross-linking of the propane side chain, especially with the Tr lignin. Alkali lignin n.m.r. spectra were considerably different from DMFL n.m.r. spectra. For example over 40% of the total bm DMFL protons were highly shielded protons but these were virtually absent in the alkali lignins. These highly shielded protons cannot be explained by the presence of ferulic and p-coumaric acids in DMFL.     

Roles of structural phenylpropanoids in forage cell wall digestion

Phenolic constituents (lignins and phenolic acids) and carbohydrates are assembled in a tight architecture which differs according to the plant species. During cell wall digestion, the hydrolysis kinetics differ between carbohydrates and seem to depend chiefly on the content and organisation of tissue phenolics. Among the phenylpropanoids, ferulic acid is released more quickly than p-coumaric acid. Lignins remain largely in the cell walls. They also undergo transformations, chiefly solubilisation as lignin-carbohydrate complexes. The limiting effect of lignins on cell wall degradation increases with increasing content. However, their effect on degradation might also depend on qualitative factors such as lignin structure and polymer organisation in walls and tissues. When various grasses (normal and selected genotypes), or grasses and legumes are compared, correlations between certain factors such as lignin uncondensed fraction, syringyl units or phenolic acids contents and cell wall degradation emerge but not clear causal relationship has been shown. Nonetheless, other structural characteristics, related to the alkali reactivity of lignins, seem to have a stronger influence on cell wall degradation. Phenylpropanoids seem to act mainly as a physical and (bio)chemical barrier to the action of the microbial enzymes. In addition, their reactivity as phenolic compounds and their hydrophobicity seem to play a role. Digestion is not limited only by phenolics. The factors that limit glycanolysis—the accessibility, crystallinity and capillary structure of cellulose and the branching of hemicelluloses—seem to have little or no effect on cell wall degradation in vivo. In contrast, other antiquality substances (tannins, cutin and silica), plant antomy, environmental factors, factors modulating microbial growth and animal physiology influence cell wall utilisation. Future research in this field should focus on the effects of phenolic structure and of cell wall and tissue organisation on carbohydrate degradation.     

A comparison of some chemical and physical properties of alkali lignins from grass and lucerne hays before and after digestion by sheep

The chemical and physical properties of alkali lignins from a grass hay and a lucerne hay were studied, before and after digestion by sheep. A greater proportion of the total grass lignins was dissolved by alkali than with the lucerne lignins but digestion increased this proportion with both species. Of the lignin dissolved by alkali, less of it was precipitated on acidification when faeces (50–70%) were compared with the undigested hays (86–92%). Extraction of lignins with boiling alkali produced a lignin with the smallest amount of amino acids but considerably altered the chemical structure as revealed by nuclear magnetic resonance (n.m.r.). Trypsin treatment of a cooler alkali extract was less effective in reducing amino acid contamination but did not alter the structure. Compared with the grass alkali lignins, the lucerne lignins had more total but fewer aromatic protons, fewer groups reacting with N-2,6-trichloro-p-benzoquinone imine, ionisable hydroxyl groups and fewer methoxyl groups. On Sephadex G-200 gel chromatography, three peaks were apparent with the lucerne, but only two with grass lignins. Both plant lignins showed large amounts of highly shielded n.m.r. protons but there were more with lucerne. Infrared spectra suggested that they were probably due to methyl groups. Digestion had very little effect on the n.m.r., ultraviolet or infrared spectra or the molecular weight distribution of the isolated lignins. It may have reduced the numbers of groups, reacting with N-2,6-trichloro-p-benzoquinone imine with the grass and two maize samples examined, but appeared to have the opposite effect with the lucerne.     

FC法测定木素和纸浆中的酚羟基

应用FC法(Folin-Ciocalteu试剂法)测定了硫酸盐木素与H2O2反应前后酚羟基含量的变化,并测定了蒸煮、氧脱木素和H2O2漂白后不同纸浆中残余木素的酚羟基含量。测定结果表明,硫酸盐木素在与碱性H2O2反应后,酚羟基含量降低,高温H2O2反应后的下降程度更为明显;浆料残余木素中酚羟基含量随浆料卡伯值的降低而下降。     

Modification of beech veneers with lignin phenol formaldehyde resins in the production of laminated veneer lumber (LVL)

Rotary cut beech (Fagus sylvatica L.) veneers were treated with four different lignin phenol formaldehyde (LPF) solutions using dimethyl sulfoxide (DMSO) as a solvent. Four of these veneers were bonded with PF adhesive to produce four-layer laminated veneer lumber (LVL). To synthesize the LPF solutions, a commercial phenol formaldehyde resin (PF resin) was individually mixed with three different technical lignins (Indulin AT, BioChoice lignin, organosolv lignin) and lignin cleavage products (LCP) at a ratio of 3:2 (60%:40%). Differential scanning calorimetry showed an increased curing temperature for the LPF resins in comparison to the PF resin. The mechanical and water-related properties of the LPF-modified LVL were shown to be similar or slightly improved compared to PF-modified LVL. Fungal degradation experiments with white-rot fungus (Trametes versicolor) and brown-rot fungus (Coniophora puteana) exhibited no significant differences in the mass loss of the LPF-modified and PF-modified samples except in one case: LVL made from veneers treated with Indulin AT exposed to the white-rot fungus. The resistance to weathering of LVL samples made from veneers treated with technical lignins was low; however, specimens treated with LCP and the reference PF resin displayed a higher resistance to weathering. It is concluded that technical lignins or LCP can, to a certain extent, be used as a substitute for crude-oil based PF resin.     

An investigation of lignin extraction from dietary fibre using acetyl bromide

Acetyl bromide is a suitable reagent for the extraction of lignins from woody material and grasses but lignins present in vegetables do not give the expected absorbance spectrum. Treatment of the vegetable fibre with either organic solvents or dilute acid or the removal of protein and hemicellulose does not alter the absorbance spectrum of lignins present in vegetables. All methods used for lignin extraction are quantitatively consistent and show only a trace of lignin to be present in vegetable fibre.     

The composition and chemiluminescence of acidolysis lignins from native Chilean hardwoods

Nothofagus obliqua (roble blanco), Nothofagus glauca (roble colorado), Lithraea Caustica (litre), Quillaja saponaria (quillay) and Cryptocarya alba (peumo), classified as hardwood species, were subjected to chemical and spectroscopic analysis. A high extinction coefficient at 280 nm correlates with the high free phenolic group content of the lignins of these species. Nothofagus glauca exhibited a high methoxyl content (19.3%) and the lower phenolic content (1.38%). The chemiluminescent behaviour of these lignins was found to be directly proportional to phenolic groups content and inversely proportional to the methoxyl content.

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Darstellung und Eigenschaften von aus Vanillin und Syringaaldehyd erhältlichen Polyestern

From vanillic and syringic acid, which are obtainable from the oxidation products of harwood lignins, polyesters were prepared by two procedures. By procedure A the respective phenolic acid was reacted with ethylene oxide to the corresponding hydroxyethyl ether, which itself was polycondensated in a second stage. By procedure B the phenolic acid was etherified with ethylene dibromide to a dibasic acid. In further stages the dibasic acid was esterified with ethylene glycol and then polycondensated. The glass transition points, melting points and molecular weights of the polyesters obtained were determined. As the results show, the polyesters differ in their melting points. The polyesters from vanillic acid exhibit relatively high melting points above 200°C, the polyesters from syringic acid are largely amorphic products.     

Chemical and Structural Analysis of Fibre and Core Tissues from Flax

Samples of flax ( Linum usitatissimum ) stems from the cultivars ‘Natasja’ and ‘Ariane’ were separated into fibre and core fractions and analysed by gas–liquid chromatographic methods, ¹³C CPMAS NMR spectrometry, histochemistry, electron microscopy and UV absorption microspectrophotometry to assist in determining the structure and composition of these cell walls in relation to quality and utilisation. Analyses from chromatography and NMR gave similar results for carbohydrate and phenolic constituents in various samples and in the lower, more mature regions of the stem. Amounts of uronic acids and xylose were lower while amounts of mannose, galactose and glucose were higher in fibre vs core fractions. Quantities of phenolic constituents were significantly higher in the core than the fibre, with groups representative of both guaiacyl and syringyl lignins; amounts of phenolic acids were low. NMR showed a low intensity signal for aromatics in fibre, and it is possible that such signals arise from compounds in the cuticle rather than the fibre. Microscopic studies indicated that aromatic constituents were present in core cell walls, cuticle of the epidermis, and cell corners and middle lamellae of some regions within the fibre tissues. The lignin in fibre appeared to be of the guaiacyl type and may be too low in concentration to be unambiguously detected by NMR. Aromatic compounds were not observed in the epidermis or parenchyma cell walls. Similar analyses of dew-retted (unscutched) samples indicated that core tissues were mostly unchanged from unretted samples. Retted fibre tissues still contained lignified cell corners and middle lamellae in some regions. The cuticle, which was associated with retted fibres, was not degraded by dew-retting fungi. Fungi removed interfibre materials in some places and at times degraded the secondary wall near the cell lumen of fibre cells. Results indicate that microspectrophotometry and histochemistry are useful to identify the location and type of aromatics in fibre cell walls.     

黑液木素制备酚醛树脂胶黏剂的研究进展

木素与酚醛树脂结构相似,可代替部分苯酚制备酚醛树脂胶黏剂。通过对木素胶黏剂的研究现状的系统归纳,介绍了黑液木素制备酚醛树脂胶黏剂的两种方式:一是木素直接替代苯酚,二是木素经活化后再与酚醛树脂结合。合理地利用造纸废液中的黑液木素不仅可以有效地处理大量的造纸黑液,而且可以降低酚醛树脂胶黏剂的生产成本,产生可持续发展的经济效益。     

Severe inhibition of maize wall degradation by synthetic lignins formed with coniferaldehyde

Although the enzymatic or ruminal degradability of plants deficient in cinnamyl alcohol dehydrogenase (CAD) is often greater than their normal counterparts, factors responsible for these degradability differences have not been identified. Since lignins in CAD deficient plants often contain elevated concentrations of aldehydes, we used a cell-wall model system to evaluate what effect aldehyde-containing lignins have on the hydrolysis of cell walls by fungal enzymes. Varying ratios of coniferaldehyde and coniferyl alcohol were polymerised into non-lignified primary walls of maize (Zea mays L) by wall-bound peroxidase and exogenously supplied H₂O₂. Coniferaldehyde lignins formed fewer cross-linked structures with other wall components, but they were much more inhibitory to cell wall degradation than lignins formed with coniferyl alcohol. This suggests that the improved degradability of CAD deficient plants is not related to the incorporation of p-hydroxycinnamaldehyde units into lignin. Degradability differences were diminished if enzyme loadings were increased and if hydrophobic aldehyde groups in lignins were reduced to their corresponding alcohols by ethanolic sodium borohydride. © 1998 Society of Chemical Industry.