Effects of Metal Ions and EDTA on Free Radical Reaction Intermediates of Laccase-catalyzed Oxidation of Wood Powder from Szemao Pine

INTRODUCTIONLaccase (EC 1.10.3.2) is multicopper oxidase catalyzing oxidation of various phenolic compounds, aromatic amines, and even certain inorganic compounds by a one-electron transfer mechanism. The electron withdrawn from the substrates is transferred via four copper atoms to molecular oxygen (Messerschmidt 1997). Laccase is very common in nature, especially in plants and fungi (Gianfreda et al. 1999). Fungal laccases participate in plant pathogenesis, pigment production and ligni…     

酶活化处理条件及其对松木纤维胶合性能的影响初探

通过测定漆酶处理纤维压制纤维板的内结合强度 ,观察到漆酶对云南思茅松木纤维的活化作用及漆酶处理对木纤维自身胶合的贡献。不同pH值条件下漆酶处理木纤维的内结合强度显著高于水处理纤维压制的板材强度。失活后的酶液与水等效 ,说明对实现木材自身胶合起作用的主要是漆酶本身。初步探索了pH值、酶剂量、底物浓度、活化处理时间等因素对胶合效果的影响。采用试验得到的较好活化条件压制的板材内结合强度可达1 0MPa以上     

Extractives of <Emphasis Type="Italic">Quercus crispula</Emphasis> sapwood infected by the pathogenic fungi <Emphasis Type="Italic">Raffaelea quercivora</Emphasis> I: comparison of sapwood extractives from noninfected and infected samples

The extracts of Quercus crispula infected by the ambrosia fungus, Raffaelea quercivora, were investigated. Phenol and tannin analyses indicated that normal sapwood (NS) contained a considerable amount of hydrolysable tannins, while infected colored sapwood (IS) contained less hydrolysable tannins and more phenols than NS. In treating pentagalloyl glucose (PGG), which is a model compound of hydrolysable tannins, with a culture medium of R. quercivora, PGG was rapidly hydrolyzed to produce gallic acid. The resulting gallic acid decreased in concentration over the subsequent cultivation period eventually disappeared. Measuring tannase and laccase activities of the culture medium of R. quercivora, tannase activity increased gradually from the beginning, while laccase activity increased rapidly at 5 days of incubation and disappeared at 8 days. An oxidative product from gallic acid treated with laccase was isolated by preparative high performance liquid chromatography, and was identified as purprogallincarboxylic acid (PGCA) by nuclear magnetic resonance spectroscopy and electron-impact mass spectrometry. PGCA was present in a 70% aqueous acetone extract of IS, and showed slight growth inhibition against R. quercivora. Part of this study was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, Japan, 2007     

Lignin-degrading activity of edible mushroom <Emphasis Type="Italic">Strobilurus ohshimae</Emphasis> that forms fruiting bodies on buried sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) twigs

Strobilurus ohshimae is an edible mushroom, and it specifically forms its fruiting bodies on buried sugi (Cryptomeria japonica) twigs. In this research, we studied lignindegrading activity of S. ohshimae. We isolated 18 strains of S. ohshimae from various regions of Japan, and determined their lignin degradation rates on sugi wood meal medium. All the strains of S. ohshimae degraded approximately 6%–12% of sugi lignin in 30 days, and these lignin degradation rates were 1.5–3 times higher than those of Trametes versicolor, which is a typical lignin-degrading fungus. Among the three main lignin-degrading enzymes, activity of lignin peroxidase and manganese peroxidase was not observed, while 4340U/g of laccase was produced in 30 days. To investigate the effect of wood species on lignin degradation by S. ohshimae, the lignin degradation rate and laccase productivity on sugi wood meal medium were compared with those on beech (Fagus crenata). In T. versicolor, both lignin degradation rate and laccase productivity were higher on beech than on sugi. Conversely, in S. ohshimae, lignin degradation rate and laccase productivity were higher on sugi than on beech. Therefore, it was suggested that coniferous lignin is not always difficult to degrade for the fungi that inhabit softwood. Part of this article presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004     

Fracture in Norway spruce wood treated with <Emphasis Type="Italic">Physisporinus vitreus</Emphasis>

Changes in the fracture behaviour of Norway spruce tonewood after fungal treatment were studied. Specimens were incubated for 6, 9 and 12 months with Physisporinus (P.) vitreus. Fracture tests were performed in a compact-tension fracture experiment set-up, and the results were compared with the morphological analysis of the degraded wood structure and transverse sections of the crack tip viewed under light and fluorescence microscopy. It was evident that both the failure load and critical stress intensity factors were reduced in wood after prolonged incubation periods. Weight losses were significantly higher in sapwood than in heartwood. With prolonged incubation periods, the frequency of unstable fracture and brittle behaviour of the wood increased. In untreated wood, cracks were initiated in the earlywood. The process involved both delamination of the cells within the middle lamellae and rupture of the cell walls, inducing a zigzag crack tip pattern. In fungally treated wood, cracks often commenced from the intersection between late- and earlywood, resulting in a straight tangential crack line. Micrographic images showed that P. vitreus was more active in the secondary walls of latewood tracheids. In this region of the wood, the cell walls were strongly degraded after 9–12 months of incubation, resulting in a reduction in tensile strength, even though the wood did not show strong features of decay at the macroscopic level.     

Mechanical properties,components and decay resistance of Populus davidiana bioincised by Coriolus versicolor

The experiment was conducted to improve the decay resistance of wood by increasing the wood preservatives uptake and penetration depth by bioincising with Coriolus versicolor. The mechanical properties of bioincised Populus davidiana sapwood were measured. The changes of wood components(lignin, cellulose and hemicellulose) were analyzed by FTIR. The staining treatment using potassium permanganate was to measure the preservatives uptake and penetration depth in wood specimens. The decay resistance of bioincised specimens impregnated with IPBC and CA was assessed against C.versicolor and Gloeophyllum trabeum using bioincised P.davidiana sapwood and untreated controls. The results showed that the mechanical properties of the bioincised specimens were changed with the incubation time and mass loss. The mechanical strength was decreased after 17 days compared to that of the specimens bioincised 13 days.The bioincised specimens demonstrated the deeper penetration depth than the un-bioincised specimens. It was significant correlation(P 0.01) between penetrationdepth and incubation time. The average value of the penetration depth of the bioincised specimens was deeper than the control groups by 158.7%. By FTIR analysis, the biodegradation ability of C. versicolor to decay cellulose and hemicellulose was weaker than that of lignin. The bioincising treatment significantly increased the wood uptake of IPBC and CA, and reached the maximum uptake value at 17 and 13 days, respectively. Summarily, the bioincising pretreatment can obviously improve the wood decay resistance against C. versicolor and G. trabeum following by introducing the IPBC and CA.     

秦岭细粘束孢产漆酶的培养条件

漆酶(Laccase,EC1.10.3.2)属于蓝色氧化酶家族的一种多酚氧化酶,是重要的木质纤维(lingocellulose)降解酶之一(Bertrand et al.,2002)。漆酶能催化降解多种芳香族化合物特别是酚类物质(Solomon et al.,1996)。由于漆酶具有430～790mV氧化还原电位(Shleev et al.,2003),能够将分子氧直接还原为水,在没有H2O2和其他次级代谢产物存在下,只要存在溶解氧,就可直接氧化底物(韩     

The influence of high defibration temperature on the properties of medium-density fiberboard (MDF) made from laccase-treated softwood fibers

This study was carried out to elucidate the effect of defibration temperature in the range 171–202°C on the properties of 12-mm thick MDF boards made without synthetic resins from softwood fibers activated by laccase treatment for the generation of phenoxy radicals on the fiber surfaces. Laccase treatment generated radicals in the fibers. An increase in defibration temperature improved the reactivity of fibers during laccase-catalyzed oxidation. The number of radicals detected in the fibers after laccase treatment in water suspension and the fiber oxygen consumption during the treatments increased with an increase in defibration temperature, while a concurrent improvement was observed in the mechanical strength and thickness swell of dry-process MDF boards made from fibers refined at different temperatures and treated with laccase in the refiner blowline. The different fiber reactivities or board properties were not due to a presence of different amounts of lignin remaining on the fiber surfaces after acetone extraction. The probable reason for them was the fact that the amount of low-molecular weight lignin, a reactive substrate for laccase, increases with increasing defibration temperature. The adhesion occurring during pressing is thus likely to involve coupling or other reactions of radicals located on adjacent fibers, whereby interfiber covalent bonds are formed.     

Assessment of decay risk of airborne wood-decay fungi

The decay risk of airborne wood-decay fungi was investigated by using an air sampler. Japanese cedar disks 7.8 cm in diameter and about 3 mm in thickness with moisture content of about 100% were placed in a “BIOSAMP” air sampler and exposed to 1000 l air. Air sampling was carried out from June to September at the same sampling site in Tsukuba, Japan. The exposed disks were then incubated for 16 weeks in a damp container kept at 26° ± 2°C. During the incubation period, wood mass loss ranged from −15 to 807 mg with a mean mass loss of 244 mg. Factors affecting mass loss were explored. Wood moisture content and ratio of heartwood area proved to be significant factors. In addition, six weather factors were found to influence mass loss. Disks that were sampled on a cloudy day showed significantly higher mean mass loss compared to those sampled on a sunny day. Subculturing of filamentous fungi from 16-week incubated disks suggested one-third of the isolated fungi produced ligninolytic enzymes.     

Laccase isoenzyme patterns of European Armillaria species from culture filtrates and infected woody plant tissues

Polyacrylamide isoelectric focusing with specific staining for laccase activity was used to characterize laccase from European Armillaria species (Armillaria ostoyae, Armillaria mellea, Armillaria gallica, Armillaria cepistipes). The enzyme was extracted from culture media either supplemented, or not, with pine sawdust, and also from Pinus pinaster naturally infected by A. ostoyae, or artificially inoculated with A. mellea and A. ostoyae. Some differences in banding patterns were found for Armillana isolates according to the species and the culture media, but a common band at pI = 3.4 was found in all the extracts tested, independently of their origin (culture filtrate or wood).     

三种树种木纤维的漆酶活化胶合

通过测定经漆酶活化处理后的思茅松、枫香和杉木纤维制成的纤维板的内结合强度,对木纤维活化后的胶合性能进行了比较.结果显示:在相同条件下进行漆酶活化处理,不同树种木纤维的自身胶合性能有一定差别,可能与其木素含量有关;不同树种木纤维进行酶活化反应时介质最佳pH值均在酸性范围,但略有差别.     

木纤维PP/PE共混物复合材料的流变和力学性能(英文)

For evaluation of the rheological and mechanical properties of highly filled wood plastic composites （WPCs）, polypropylene/polyethylene （PP/PE） blends were grafted with maleic anhydride （MAH） to enhance the interfacial adhesion between wood fiber and matrix. WPCs were prepared from wood fiber up to 60 wt.% and modified PP/PE was blended by extrusion. The rheological properties were studied by using dynamic measurement. According to the strain sweep test, the linear viscoelastic region of composites in the melt was determined. The result showed that the storage modulus was independent of the strain at low strain region （〈0.1%）. The frequency sweep resuits indicated that all composites exhibited shear thinning behavior, and both the storage modulus and complex viscosity of MAH modified composites were decreased comparing to those unmodified. Flexural properties and impact strength of the prepared WPCs were measured according to the relevant standard specifications. The flexural and impact strength of the manufactured composites significantly increased and reached a maximum when MAH dosage was 1.0 wt%, whereas the flexural modulus after an initial decreased, also increased with MAH dosage. The increase in mechanical properties indicated that the presence of anhydride groups enhanced the interracial adhesion between wood fiber and PP/PE blends.     

Lignification and peroxidase in tension wood ofEucalyptus viminalis seedlings

Seedlings ofEucalyptus viminalis were grown for 50 days with their stems bent so tension wood would form. Every 10 days the lignin content, monomeric composition, and peroxidase activity in the tension wood were compared with those in the lower side (opposite wood) and in vertically grown controls. The lignin content in the developing tension wood started to decrease after 10 days of bending and kept decreasing for 50 days, whereas those in control plants and opposite wood remained almost unchanged. The yields of syringaldehyde from tension wood by nitrobenzene oxidation increased, and consequently the syringyl/ guaiacyl ratio of the lignin was higher in tension wood than in opposite wood and control plants. The peroxidase ionically bound to the cell walls (IPO) catalyzed oxidation of guaiacol and syringaldazine. The syringaldazineoxidizing activity of IPO from tension wood increased, whereas the activities of IPO from opposite wood and control plants did not show any marked change. In tension wood the increase in syringaldazine-oxidizing activity of IPO was consistent with an increase in the syringaldehyde yield. This suggests that IPO contributes to syringyl lignin deposition as other enzymes involved in the monolignol biosynthesis do in tension wood formation.This study was presented at the 50th Annual Meeting of the Japan Wood Research Society, Kyoto, April 2000     

Effect of coniferyl alcohol addition on removal of chlorophenols from water effluent by fungal laccase

The effect of coniferyl alcohol on removal of chlorinated phenols from a water environment byRhizoctonia praticola andCerrena unicolor laccases was studied. At optimal conditions in which 7 mM coniferyl alcohol and laccase were added to chlorinated phenols over 20h, about 34% of the radioactivity of 4-chlorophenol, 57% of 2,4-dichlorophenol, 66% of 2,4,5-trichlorophenol, and 85% of pentachlorophenol were removed from the supernatants, compared to the level without laccase activity. After 12-h incubation periods at the optimal concentrations of coniferyl alcohol and laccase (added simultaneously), the fast first phase of chlorophenol removal was complete in 1 h, and eventually coniferyl alcohol enhanced the removal of 4-chlorophenol by 40%, 2,4-dichlorophenol by 54%, 2,4,5-trichlorophenol by 60%, and pentachlorophenol by 76%.     

Variation in growth of Heterobasidion annosum among clones of Picea abies incubated for different periods of time

Thirty‐seven 4‐year‐old clones of Picea abies were inoculated with one isolate of the S intersterility group of Heterobasidion annosum in a greenhouse. The dehardened cuttings were organized in three different groups with four to six ramets in each group. All groups were inoculated on the same day shortly after shoot elongation. The groups were sampled for H. annosum growth after 34 (group 1), 83 (group 2) and 182 days (group 3), respectively. Measured parameters were cutting height and diameter, vigour index of the cuttings, infection incidence, mortality rate and fungal growth in the stem. The height of the cuttings was almost constant during the 6 months of incubation, whereas the diameter increased by about 10% during the same period. The proportion of living cuttings containing H. annosum decreased with time (99.5, 93 and 67% infection in groups 1, 2 and 3, respectively) and differed significantly among clones in group 3. Mortality rate increased with time (0.5, 22 and 37% mortality in groups 1, 2 and 3, respectively) and differed significantly among clones in groups 2 and 3; the same clones being most susceptible for both times. Mean fungal growth into the wood was significantly different among groups and among clones within each group. The ranking position for fungal growth was similar in the three groups. The results indicate that frequency of infection success, mortality rate and fungal growth are clone‐dependent factors. Broad sense heritability varied between 0.08 and 0.25 for fungal growth and lesion length at the three incubation periods. Infection success frequency was initially not different among clones but in the longer incubation periods there were significant differences among clones, indicating differences in resistance. Fungal growth in wood differed among separate host clones irrespectively of the length of inoculation period. The vigour of the cuttings seem to influence the length of fungal extension only in the initial stage of the infection.     

红平菇木质素降解酶系统漆酶、锰过氧化物酶及木质素过氧化物酶的检测

木材白腐菌在分解木质素的过程中会产生非特异性的分解木质素结构的酶系统,这些酶系统主要包括细胞外过氧化物酶[锰过氧化物酶(manganese peroxidase,MnP)、木质素过氧化物酶(lignin peroxidase,LiP)]和细胞外酚氧化酶[漆酶(laccase)].因此,在生物修复方面,白腐菌能够有效地降解废水和土壤中难被降解的多氯联苯、多环芳烃、DDT、染料、炸药和其他氯化物、叠氮化合物等.     

利用木质剩余物与废纸制备纸浆模塑材料的比较

对三种缓冲材料及利用木质剩余物一次纤维和废纸再生纤维制备纸浆模塑材料工艺过程进行对比。并利用扫描电镜观察热压干燥和真空干燥成型时,木质剩余物一次纤维与废纸再生纤维纤维形态结构的变化。结果表明：木质剩余物一次纤维纤维长度、宽度较废纸再生纤维得均匀,废纸再生纤维纤维表面有所损伤,纤维间的空隙增加,会导致与纤维间结合力有关的抗张强度、耐破强度和耐折强度等物理性能地下降。在生产实践中,要根据包装物的具体情况,选择合适的纸浆模塑原材料。     

Stem tangential strain on the tension wood side of <Emphasis Type="Italic">Fagus crenata</Emphasis> saplings

The tangential strain on the inner bark surface of Fagus crenata sapling stems was continuously measured using strain gauges. The total strain increased daily, increasing at night and decreasing during the day. When tension wood was induced by artificial inclination, the strain increased more on the upper side than on the lower side; and the increment in the strain at night was larger on the upper side than on the lower. The change in tangential strain on the inner bark surface arose from changes in the water content and the volume of differentiating cells. Differentiating tension wood fibers appear to contain more water and to expand more at night than differentiating normal wood fibers. We can determine whether tension wood is formed from the tangential strain during growth.     

Persistence of some pine pathogens in coarse woody debris and cones in a Pinus pinea forest

The persistence of Sphaeropsis sapinea, Leptographium serpens and Heterobasidion annosum s.s. in artificially inoculated pine branch pieces (S. sapinea and L. serpens) and wood blocks (L. serpens and H. annosum s.s.) was investigated in order to discuss the alternative of leaving coarse woody debris in stands of Italian stone pine (Pinus pinea). Also, natural colonization by S. sapinea of pine cones of different ages was assessed. Methods used for inoculating branch pieces and wood blocks were highly effective for all fungi. Type of a forest stand in which branch pieces and wood blocks have been incubated did not affect the persistence of the pathogens in the inoculated samples. For branch pieces, the success of re-isolation of L. serpens dropped as the sample incubation time increased, while S. sapinea was always successfully (100%) re-isolated (even 12 months after the inoculation). L. serpens and H. annosum s.s. were re-isolated from most of the buried wood blocks (from more than 95% samples) up to 3 months following the inoculation. Of the observed P. pinea cones (in most cases, more than 2 years old), 74% were naturally infected byS. sapinea. All three investigated pathogens were able to survive in dead plant tissues for long periods of time (at least for several (3–12) months). The persistence of these pine-pathogenic species in dead plant material questions the feasibility of leaving coarse woody debris in managed Italian stone pine forests meant for landscape conservation and leisure activities.