• Journal of the Korean Wood Science and Technology
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• v.28 no.4
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• pp.29-40
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• 2000

Highly purified Cerrena unicolor laccase (benzenediol:oxygen oxidoreductase, EC 1.10.3.2) caused the demethoxylation of milled wood lignin and several lignin related substances. The constitutive form of the enzyme produced extracellularly by C. unicolor fermenter culture was isolated and purified by ion-exchange chromatography on the DEAE-Toyopearl column and by affinity chromatography on a ConA-Sepharose and Syringyl-AH-Sepharose 4B columns. The enzyme was further immobilized on functionalized porous glass (CPG) and keratin coated CPG. The demethylating activity was monitored both by estimation of released methanol and by detection of the level of methoxyl groups (also in some water miscible solvents) after incubation of lignin materials with laccase preparations (free and immobilized). The effects of the incubation time and temperature on the demethoxylating activity of immobilized laccase preparations were also studied.

• Journal of the Korean Wood Science and Technology
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• v.24 no.1
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• pp.68-74
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• 1996

Present work was undertaken to investigate the origin of milled wood lignin(MWL) in the wood cell wall using immunocytochemical techniques, which can provide the information on the localization of specific antigens(MWL in the present study) to be examined. Spruce MWL dissolved in DMSO and emulsified with Freund adjuvant was injected directly into the mouse spleen. The animals were boostered at two-week intervals after the initial immunization. Blood samples were purified in standard procedures. The characteristics of antibodies against MWL were tested by indirect ELISA. Visualization of MWL was carried out using conventional indirect immunogold-labelling methods on the ultrathin sections of spruce wood. Immuno-TEM observations showed that the immunogold probes were selectively attached to secondary cell walls of spruce wood. The most intense labelling was frequently observed in the S2 layer. In contrast, gold labelling in the lignin-rich regions, such as middle lamella and cell corner was not found. The immuno-TEM provides an indication that spruce MWL originates from the S2 layer.

• Journal of Korean Society of Forest Science
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• v.45 no.1
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• pp.62-67
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• 1979

Ultraviolet microscopy of ultrathin sections of wood has proved to be one of the useful means for determining the lignin distribution in the various regions of the cell wall. Also, spectral approach and quantitative analysis of isolated compound middle lamella fraction from birch xylem have revealed that the lignin associated with the vessel secondary wall and middle lamella is composed predominantly of gualacylpropane units. Lignin deposited in the fiber and ray parenchyma secondary walls is composed mostly of syringylpropane units. The middle lamella lignin around fibers and ray cells contains both guaiacyl and syringyl propane quits. On the basis of the results above, this research was carried out to clarify the origin of milled wood lignin (MWL) by analysing the chemical characteristics of ML MWLs extracted at various milling stages. The amount of phenolic hydroxyl-, ${\alpha}$-carbonyl-, and methoxyl-group in the MWL's increases the milling time. And progressive mining contributes to the merease of ratio of syringylaldehyde to vanillin(S/V ratio) after nitrobenzene oxidation of MWL. Accordingly, It could be concluded that milled wood lignin extracted at the initial milling stage derives from compound middle lamella region of cell wall, whereas, with progressive milling, lignin of secondary wall of fiber is introduced gradually to milled wood lignin. These results are suggesting that heterogeneous chemical structure of lignins in hardwood exists. Although milled wood lignin at the initial stage seems to have lower molecular weight in comparison with milled wood lignin extracted at final milling stage from the result of Gel-filtration curves, further study would be required on molecular weight distribution of milled wood lignin in future.

• Journal of the Korean Wood Science and Technology
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• v.39 no.1
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• pp.86-94
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• 2011

Pyrolytic lignin was obtained from biooil produced with yellow poplar wood. Fast pyrolysis was performed under various temperature ranges and residence times using fluidized bed type reactor. Several analytical methods were adopted to characterize the structure of pyrolytic lignin as well as the effect of pyrolysis temperature and residence time on the modification of the lignin. The yield of pyrolytic lignin increased as increasing pyrolysis temperature and decreasing residence time of pyrolysis products. The molecular weight of pyrolytic lignin determined by gel permeation chromatography (GPC) was approximately 1,200 mol/g, which was approximately a tenth of milled wood lignin (MWL) purified from the same woody biomass. Based on analytical data, demethoxylation and side chain cleavage reaction were dominantly occurred during fast pyrolysis.

• Journal of the Korean Wood Science and Technology
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• v.40 no.3
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• pp.194-203
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• 2012

1-Ethyl-3-methylimidazolium acetate known as efficient biomass pretreatment reagent was used for the extraction of lignin from rigida pine wood (pitch pine), which was called to ionic liquid lignin (ILL), and chemical structural features of ILL were compared with the corresponding milled wood lignin (MWL). The amounts of phenolic hydroxyl groups (Phe-OH) was determined to 10.0% for ILL and 7.2% for MWL, respectively, where those of methoxyl groups (OMe) were 4.9% for ILL and 11.0% for MWL, respectively. The weight average molecular weight (Mw) of ILL (3,995) were determined to ca. 1/2 of that of MWL (8,438) and polydispersity index (PDI: Mw/Mn) suggested that the lignin fragments were more uniform in the ILL (PDI 1.36) than in the MWL (PDI 2.64). The temperature (Tm) corresponding to maximum decomposition rate (Vm) of ILL ($306.6^{\circ}C$) was ca. $35^{\circ}C$ lower than that of MWL ($341.9^{\circ}C$), suggesting that ILL was thermally unstable than MWL, as evidence from the lower Tm for ILL. Moreover, the structural characteristics of ILL and MWL were confirmed by spectroscopic analyses (FT-IR and $^{13}C$-NMR), and these results indicated ionic liquid (1-ethyl-3-methylimidazolium acetate) was chemically or physically bound to ILL.

• Journal of the Korean Wood Science and Technology
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• v.38 no.3
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• pp.213-222
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• 2010

In this study, chemical compositions - holocellulose, lignin and monomeric sugars - were characterized with two poplar wood cell walls, one of which was grown at normal condition (CPW) and the other was genetically modified by antisence suppression of CCoAOMT gene expression (ACPW). Milled wood lignins were isolated from CPW and ACPW and subjected to methoxyl group, DFRC, Py-GC/MS, GPC, $^{13}C$-NMR analysis, respectively. There were few differences in holocellulose contents in both cell walls, which were determined to 81.6% in CPW and to 82.3% in ACPW. However, lignin contents in ACPW was clearly decreased by the suppression of CCoAOMT gene expression. In CPW 21.7% of lignin contents was determined, while lignin contents in ACPW was lowered to 18.3%. The relative poor solubility of ACPW in alkali solution could be attributed to the reduction of lignin content. The glucose contents of CPW and ACPW were measured to 511.0 mg/g and 584.8 mg/g and xylose contents 217.8 mg/g and 187.5 mg/g, respectively, indicating that suppression of CCoAOMT gene expression could be also influenced to the formation of monomeric sugar compositions. In depth investigation for milled wood lignin (MWL) isolated from both samples revealed that the methoxyl contents at ACPW was decreased by 7% in comparison to that of CPW, which were indirectly evidenced by $^{13}C$-NMR spectra and Py-GC/MS. According to the data from Py-GC/MS S/G ratios of lignin in CPW and ACPW were determined to 0.59 and 0.44, respectively. As conclusive remark, the biosynthesis of syringyl unit could be further influenced by antisense suppression of CCoAOMT during phenylpropanoid pathway in the plant cell wall rather than that of guaiacyl unit.

• Journal of the Korean Wood Science and Technology
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• v.33 no.6 s.134
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• pp.79-86
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• 2005

Aspen wood (Populus tremuloides, L.) was biotreated with Ceriporiopsis subvermispora for 1, 2, 4, and 6 weeks to observe the physical/chemical modification of wood components. Milled wood lignins (MWLs) isolated from each decayed wood were analyzed by gel permeation chromatography (GPC) and nitrobenzene oxidation (NBO). As fungal treatment was progressed, lignin contents continuously decreased up to 20% after 6-week treatment. The lignin polymer could be fragmented to low-molecular phenolics, which make an enhancement of alkali solubility. Holocellulose contents were not affected severely during the period of fungal treatment, only reduction of 5~6% compared to the control. Xylose contents were decreased gradually from 23.4% to 18% after 6 weeks, whereas alpha-cellulose remained almost unchanged. Gel permeation chromatography (GPC) indicates that molecular weight of lignin undergoes a slight decrement for 4 weeks of fungal treatment. Nitrobenzene oxidation revealed that total yield of NBO products of lignins were lowered ca 20% after fungal treatment. Sum of syringaldehyde and syringic acid are remarkably decreased. However, increment of sum of vanillin and vanillic acid was surprisingly observed. These results work as indirect evidence that a specific lignolytic reaction, maybe selective demethoxylaytion of S-lignin, can occur during fungal treatment of aspen wood by C. subvermispora.

• Journal of the Korean Wood Science and Technology
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• v.37 no.4
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• pp.372-380
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• 2009

After supercritical water treatment of poplar wood meals (passed through 60 mesh) for 60s between 325 and $425^{\circ}C$ at the fixed pressure at $220{\pm}10atm$, some solid residues were present in the degradation products. They mainly consisted of chemically modified lignin and fibrous materials. Glucose and xylose were identified as main sugar components of fibrous materials, and the highest ratio of glucose/xylose was achieved at the highest reaction temperature. As reaction temperature was elevated, the portion of fibrous materials decreased in the solid residues, while lignin was further accumulated. The H : G : S ratio of lignin in solid residues was estimated by analytical pyrolysis. Irrespective of reaction temperatures, the H:G:S ratios were not significantly changed in the lignin in solid residues. Compared to poplar milled wood lignin (MWL), it was remarkable that H type monomers were further lowered, while portion of S type monomers increased. The amount of G type monomers were relative stable. In presence of HCl catalyst, lowering H type as well as enhancing S type was further distinguishable. According to the result of nitrobenzene oxidation (NBO), ca. 265 mg of vanillin and syringaldehyde was yielded from poplar MWL as main products. However, remarkably reduced amount of NBO products were determined from solid residues by raising operating temperature as well as by the addition of HCl catalyst. These results strongly indicate that $\beta$-O-4 linkage could be easily cleaved during supercritical water treatment, so that the lignins in the solid residues seem to be condensed phenol polymers, which are mainly formed by carbon-carbon linkages rather than $\beta$-O-4 linkage.

• Journal of the Korean Wood Science and Technology
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• v.38 no.3
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• pp.223-229
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• 2010

In this study diverse dehydrogenative polymers (DHPs) were synthesized with three precursors of native lignin [p-coumaryl alcohol (PCA), coniferyl alcohol (CA), sinapyl alcohol (SA)] in the presence of horseradish peroxidase (HRP, EC. 1.11.1.7)/$H_2O_2$. To compare the structural features between DHPs and native lignin, the DHPs as well as pine/poplar milled wood lignins were simultaneously subjected to gel permeation chromatography (GPC) to determine average molecular weights and derivatization followed by reductive cleavage (DFRC) to investigate the frequency of ${\beta}$-O-4 linkage. The highest yield of DHP was measured to 71% when CA was solely injected (G-DHP) and the yield of H-DHP was 42%. However, single injection of SA could not form any polymer in this system. The average molecular weights of DHPs were ranged between 3,000~4,700, which were only 1/2 fold compared with that of pine MWL (G-type lignin: Mw 7,340) and 1/3 scale compared with that of poplar MWL (GS-type lignin: Mw 13,250). DFRC analysis revealed that the formation of ${\beta}$-O-4 linkage during dehydrogenative polymerization was the highest in the GS-DHP with ca. 502 ${\mu}mol$/g, which was, however, remained to only 50% compared to that in poplar MWL (1107 ${\mu}mol$/g). The ${\beta}$-O-4 linkage was estimated to ca. 286 ${\mu}mol$/g In the G-DHP, which was twice as much as that of H-DHP(127 ${\mu}mol$/g). Similar to GS-DHP, only half amount of ${\beta}$-O-4 linkage, compared to pine MWL, was formed during in vitro polymerization of CA by horseradish peroxidase/$H_2O_2$.

• Journal of the Korean Wood Science and Technology
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• v.16 no.4
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• pp.3-9
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• 1988

Infrared (IR) spectroscopic techniques for the analysis of wood samples and the absorbance spectra of solid woods were presented. KBr pellets were prepared by throughly mixing approximately 300 mg of dried KBr and 1 mg of finely milled wood powder extracted with ethanol-cyclohexane previously. This mixture was made into a transparent disc by means of a pellet-making die (10 ton/$cm^2$ for 10 min). This IR techniques were applied for the analysis of archaeological wood samples. The most notable difference in the IR spectra between the recent and the archaeological waterlogged woods is that the absorption band centered at $1,730cm^{-1}$ was significantly diminished in the waterlogged ones. Total loss of absorption in $1,730cm^{-1}$ might be mainly due to the result of hemicellulose degradation. Another feature indicated by IR spectral comparision are that the degraded waterlogged wood samples showed 1) the increased intensity of the 1,600, 1,500 and $1,270cm^{-1}$ due to the residual lignin and the increased intensity at 1,470 and $1,425cm^{-1}$ due to the degradation of hemicellulose and 2) to the emergence of single band around $1,050cm^{-1}$ instead of three bands at 1,110, 1,060 and $1,040cm^{-1}$ in recent wood due to the degradation of cellulose crystalline. It was revealed from the IR examinations that the first change of wood in the waterlogged situation was the lysis of hemicellulose and the second the lysis of cellulose. It was also suggested that IR spectroscopy could serve a fast method for the investigation on the chemical characteristics of archaeological wood samples.