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.     

Characterization of trembling aspen wood (Populus tremuloides L.) degraded with the white rot fungus Ceriporiopsis subvermispora and MWLs isolated thereof

Trembling aspen wood (Populus tremuloides L.) was treated with white rot fungus Ceriporiopsis subvermispora for 1, 2, 4, and 6 weeks. As fungal decay advanced, lignin contents were decreased gradually up to ca. 27.5% (based on the Klason residues of the control) after 6 weeks. Alkali solubility of cell wall residues was increased until 4 weeks of fungal treatment, but additional treatment did not cause any effects. Milled wood lignins (MWLs) were isolated from the decayed woods by Björkman’s procedure and subjected to thioacidolysis and analytical pyrolysis to investigate the modification of lignin structures during fungal degradation. Thioacidolysis revealed that the yields of trithioethylated C₆C₃ monomers, as a parameter for frequency of β-O-4 linkages in lignin, were substantially reduced (-20%, based on the control) in MWLs isolated from decayed woods. Analytical pyrolysis revealed that the relative amounts of coniferyl alcohol and sinapyl alcohol in the pyrolysates were lowered dependent on the biodegradation time, whereas an elevation of C₆C₁ and C₆C₂ pyrolytic phenols was observed. The results from both analytical methods strongly suggested that β-O-4 linkages were cleaved by C. subvermispora. Specially, degradation of syringyl-type lignin seems to be preferred.     

A comparative study of hard and softwood lignins alterations during treatment in dioxane-water-HCl agent

The objective of the presented contribution was a study of the lignin alterations occurring in the process of the acidolytic delignification of chosen representatives of soft and hard wood species (maple and spruce) in dioxane-water-HCl mixture. All lignin samples were characterised by yield or recovery, molecular weights, as well as by GLC data of corresponding nitrobenzene oxidation products. In order to obtain more information about condensation and degradative reactions of lignin under conditions of acidolytic treatment a simulation of the process of lignin isolation was performed, too. Samples of dioxane lignins obtained after 90 min. of isolation at 82.5 and 98 °C were therefore recooked during 360 min. under identical conditions as applied in the lignin isolations. Concentrations of lignin in the reaction mixture were 1 and 10% thus approximately corresponding the lignin concentration inside and outside of the wood meal during the lignin isolation. The obtained experimental data point out the course of both degradative and condensation reactions. Maple wood lignin undergo deeper degradation than those of spruce under the same reaction conditions. The decreased yields of lignin oxidation products with the time of acidolytic treatment and after recooking hint at the formation of new linkages among building units of lignin thus modifying its macromolecules. The intensity of occurring alterations of examined lignins is not only dependent of time and temperature, but also of the concentration of lignin.     

黄孢原毛平革菌对大豆秸秆木质素降解的FTIR表征

研究黄孢原毛平革菌(Phanerochaete chrysosporium Burdsall)对大豆秸秆中木质纤维素的降解条件。结果显示：5g秸秆粉中添加合成培养液12mL、接种量为0.8mL(孢子悬液浓度8.6×106CFU/mL)、培养基初始pH4.5,发酵10d后,木质素降解率为45.96%。并采用傅里叶红外光谱分析(Fourier transform infrared spectroscopy,FTIR) 检测降解后秸秆中官能团的变化,发现与木质素相关的谱峰(1099cm-1、1057～1038cm-1)相对强度减小,与苯环相关的谱峰(1643～1608cm-1、1510～1508cm-1)相对强度增加,表明部分大分子木质素裂解成小分子木质素或木质素单体,对比其他谱峰相对强度的变化,发现木质素中苯环等环状化合物含量减少。     

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.     

Analytical pyrolysis and FTIR spectroscopy of fossil Sequoiadendron giganteum (Lindl.) wood and MWLs isolated hereof

Recent and fossil woods from the genus of Sequoiadendron, as well as their milled wood lignins were investigated by means of FTIR spectroscopy and pyrolysis GC/MS technique. Though almost same in appearance the differences in band intensities of FTIR-spectra reveals the distinctly increased aromatic character and the oxidation in fossil specimens. More detailed information about the changes in old wood were obtained by analytical pyrolysis, which delivers degradation products with shortened side chains and more saturated units relating to fossil lignin.     

嗜碱性木素降解菌降解能力的初步研究

通过比较不同嗜碱细菌对麦草中木素、纤维素和半纤维素的降解率, 并比较不同菌株的产酶及酶活情况, 筛选出了木素降解能力较强, 而纤维素和半纤维素降解能力相对较弱的嗜碱性木素降解菌。在ｐＨ≈１０－４的条件下培养８ｄ 后, 菌株６ 降解了３２－３７％ 的麦草木素, 而纤维素和半纤维素分别降解了２１－４８％ 和２２－６９％ 。这与其产酶情况基本一致, 该菌株的酶活最高, 分别为ＭｎＰ２７１－３０Ｕ／Ｌ和ＬｉＰ４１－９４Ｕ／Ｌ。     

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.     

秸秆基质中木质素白腐菌降解的SEM特征观察

研究了黄孢原毛平革菌(PhanerochaeteChrysosporiumBurdsall)对大豆和稻草秸秆中木质素的降解条件及降解效果。结果显示:在各5.0g秸秆中,加入合成培养液12mL、菌悬液浓度8.6×106cfu/mL时,种子添加量为0.8mL、培养基pH4.5,降解10d,测得木质素降解率分别为大豆秸秆49.91%、稻草秸秆49.50%。利用SEM对发酵前后秸秆表观结构进行观察,得到黄孢原毛平革菌对大豆和稻草秸秆有明显的降解效果。经黄孢原毛平革菌降解10d的大豆和稻草秸秆表面空穴增多、增大,甚至相互连通,降解效果明显。     

不同Cu离子浓度和pH值对云芝降解秸秆中木质素和纤维素的影响

云芝(Coriolus versicolor)对秸秆中木质素与纤维素的降解情况受不同Cu2+浓度和pH值条件的影响,设定不同条件可以提高木质素的降解率,降低纤维素的降解率,以更好地保存纤维素的完整性,实现秸秆中主要成分的分离。通过均匀设计法,设定不同的Cu2+浓度环境X1(0、1.0、2.0、4.0、6.0mmol/L)和不同pH值条件X2(2.2、2.8、3.4、4.0、4.6),测定相应条件下云芝的菌丝生长情况、木质素酶活力、纤维素酶活力以及秸秆中木质素和纤维素的降解率。结果表明：木质素的降解率与Cu2+浓度呈正相关,与pH值呈负相关;纤维素的降解率与pH值呈正相关,与Cu2+ 浓度呈负相关。通过试验所得方程预测得知,Cu2+浓度6mmol/L、pH2.2是优化的云芝降解、分离秸秆中主要成分的条件,在该条件下,由所得方程可知秸秆中木质素的降解率为16.1%,纤维素的降解率为27.7%。     

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.     

Structural changes in wheat straw components during decay by lignin-degrading white-rot fungi in relation to improvement of digestibility for ruminants

Structural changes and resulting in-vitro dry matter digestibility (IVDMD) were determined during the course of solid-state fermentation of wheat straw using the lignin-degrading white-rot fungi Sporotrichum pulverulentum, Pycnoporus cinnabarinus and Cyathus stercoreus. The first fungus grew very rapidly on straw but degraded hemicelluloses and cellulose non-selectively resulting in very low IVDMD increases (50% after 35% weight loss). P. cinnabarinus and C. stercoreus preferentially degraded hemicelluloses achieving high improvements in IVDMD (maximum increase of 63 and 94%, respectively) with limited dry weight losses (12 and 18% after 7 and 13 days, respectively). The three fungi exhibited some selectivity among the individual hemicellulose components: O-acetyls were removed essentially at the same rate as xylan, while uronic acids accumulated as incubation proceeded. Conversely, the arabinose content decreased rapidly, especially with C. stercoreus and P. cinnabarinus, suggesting that removal of this pentose was partly responsible for digestibility improvement. Esterified phenolic acids were rapidly degraded during the first stages of decay by all three fungi although P. cinnabarinus and C. stercoreus degraded ferulic acid faster than p-coumaric acid. Lignin was preferentially degraded compared to polysaccharides by all three fungi. The amount of lignin removed, as determined by Klason, correlated well with IVDMD improvement (r=?0.97), while acid detergent lignin (ADL) showed a lower correlation (r=-0.86). Acidolysis yields of decayed lignin pointed to preferential degradation of β-O-4 ether linked units by the fungi. Syringyl units were removed faster than guaiacyl units only after 5 to 10% weight loss was obtained.     

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.     

Fungal pretreatment of wheat straw: Effects on the biodegradability of cell walls,structural polysaccharides,lignin and phenolic acids by rumen microorganisms

Disappearance of cell wall components of untreated straw and straw treated with the ligninolytic white-rot fungi Phanerochaete chrysosporium, Dichomitus squalens and Cyathus stercoreus were determined during the course of rumen digestion of samples in nylon bags. The first fungus degraded hemicelluloses and cellulose non-selectively, adversely affecting the digestion rate of crude cell walls. Dichomitus squalens and C. stercoreus preferentially degraded hemicelluloses and lignin, affording cell wall degradation rates 1.5 times higher than in native straw. Furthermore, the extent of cell wall digestion was also significantly enhanced. Both strains improved the extent of cellulose digestion, whereas the potentially degradable xylan fraction remained unchanged. Polysaccharide digestion rates were influenced in different ways depending on the strain tested: straw degraded by C. stercoreus showed an increase in cellulose digestion rate by 50%, whereas residual arabinose units were slowly degraded. Xylan was degraded 1.8 times faster in straw decayed' by D. squalens, while cellulose digestion remained unchanged. Phanerochaete chrysosporium depressed both xylan and cellulose digestion rates. Fungal-treated lignins were solubilised in the rumen faster than in untreated straw, whereas only treatment by C. stercoreus resulted in higher lignin losses. Esterified phenolic acids were extensively degraded by all three fungi. Residual ferulic and p-coumaric acids accumulated during rumen digestion, although only the former decreased in the original straw.     

高氧气调包装对绿芦笋木质化的影响

以空气包装为对照,研究了60% O2+20% CO2+20% N2、80% O2+20% CO2 和100% O2 高氧气调包装的绿芦笋在(4 ± 1)℃贮藏28d 期间失重率、叶绿素含量、木质素含量、苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)活性的变化。结果表明：高氧气调包装可以减缓绿芦笋的失重,抑制叶绿素的降解、木质素含量的上升,抑制PAL和POD 酶活性的上升。80% O2+20% CO2 气调包装贮藏条件下绿芦笋失重较小、色泽保持良好、木质化程度较低。该条件可较好地延长绿芦笋货架期并保持其良好的品质。     

Bestimmung der phenolischen Hydroxylgruppen in Ligninen und Ligninfraktionen durch Aminolyse und FTIR-Spektroskopie

The quantitative analysis of phenolic OH groups is of interest for the characterization of isolated lignins when evaluating the extent to which lignin has been changed. While aminolysis is a very accurate if elaborate chemical method, the evaluation of the aliphatic and aromatic IR ester bands of acetylated lignins at 1745 and 1765 cm^?1 permits a quick, easy and reliable determination of the phenolic hydroxyl groups. The excellent correlation between the two methods was proved for 35 different lignins and lignin fractions of different molecular weights.     

Pyrolytic and hydrogenolytic degradation studies on lignocellulosics,pulps and lignins

Wood and MWL's from beech, spruce, bamboo as well as teak wood and teak-HCl-lignin were subjected to analytical pyrolysis (Py), using the off-line approach, and to hydrogenolysis (Hy). Gas chromatography-mass spectrometry (GCMS) was used for product identification and assignment as derivatives from 4-hydroxy-phenyl-propane, guaiacylpropane and syringylpropane basic units followed by capillary gas chromatography (GC) for quantitative determination of the phenolics. The results are compared with those of nitrobenzene oxidation and quantitative FTIR-spectroscopy. Py-GC and Hy-GC experiments were also used for the characterization of residual lignins in kraft and alkaline-sulphite-anthraquinone-methanol (ASAM) pulps from beech, pine and sugar cane bagasse. Analytical pyrolysis of biomass, both in on-line and off-line approach, is well suitable for lignin classification even without previous lignin isolation. Hydrogenolysis is more recommendable for the characterization of residual lignins in pulps.     

漆酶处理对杨木APMP木素分子质量的影响

研究了漆酶和漆酶/介体体系处理对三倍体毛白杨APMP性能的影响.采取酶解-弱酸解两段法分别从APMP原浆和生物处理后的纸浆中分离木素.为了使木素在凝胶渗透色谱流动相四氢呋喃中能完全溶解,对所有试样进行了卞基化处理.利用凝胶渗透色谱分析仪获得了木素的分子质量分布曲线,并计算出数均分子质量和质均分子质量,进而比较了漆酶和漆酶/介体体系处理的效果和差异.     

不同氧化剂对禾草类碱木质素结构变化的影响

研究了禾草类碱木质素在O2、H2O2、O3、O2/H2O2、O3/H2O2等不同氧化剂氧化时其结构的变化规律,为更好地利用禾草类碱木质素提供理论依据和工艺方法。研究结果表明,以O2/H2O2氧化禾草类碱木质素时,高用碱量可促进木质素高分子组分氧化降解,分子质量均一化,酚羟基和羧基增加较多,脱甲氧基反应也较强烈;低用碱量时,木质素氧化降解少,并伴有缩合反应;H2O2可促进碱木质素氧化降解,促进酚羟基、羧基的产生和脱甲氧基反应;在O3氧化适宜条件下,添加H2O2可使O3氧化反应缓和,木质素分子发生缩合并均一化,生成较多的酚羟基、羧基和羰基。     

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.