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Ceriporiopsis subvermispora is a white-rot fungus with a high specificity towards lignin mineralization when colonizing dead wood or lignocellulosic compounds. Its lignocellulose degrading system is formed by cellulose hydrolytic enzymes, manganese peroxidases, and laccases that catalyze the efficient depolymerization and mineralization of lignocellulose. To determine if this metabolic specialization has modified codon usage of the lignocellulolytic system, improving its adaptation to the fungal translational machine, we analyzed the adaptation to host codon usage (CAI), tRNA pool (tAI, and AAtAI), codon pair bias (CPB), and the number of effective codons (Nc). These indexes were correlated with gene expression of C. subvermispora, in the presence of glucose and Aspen wood. General gene expression was not correlated with the index values. However, in media containing Aspen wood, the induction of expression of lignocellulose-degrading genes, showed significantly (p &lt, 0.001) higher values of CAI, AAtAI, CPB, tAI, and lower values of Nc than non-induced genes. Cellulose-binding proteins and manganese peroxidases presented the highest adaptation values. We also identified an expansion of genes encoding glycine and glutamic acid tRNAs. Our results suggest that the metabolic specialization to use wood as the sole carbon source has introduced a bias in the codon usage of genes involved in lignocellulose degradation. This bias reduces codon diversity and increases codon usage adaptation to the tRNA pool available in C. subvermispora. To our knowledge, this is the first study showing that codon usage is modified to improve the translation efficiency of a group of genes involved in a particular metabolic process.

Aim The present study evaluated differences between two strains of Ceriporiopsis subvermispora on improving the nutritive value and in vitro degradability of wheat straw. Methods and Results Wheat straw was treated with the fungi for 7 weeks. Weekly samples were analyzed for ergosterol content, in vitro gas production (IVGP), chemical composition and lignin degrading enzyme activity. Ergosterol data showed CS1 to have a faster initial growth than CS2 and reaching a stationary phase after 3 weeks. The IVGP of CS1-treated wheat straw exceeded the control earlier than CS2 (4 vs 5 weeks). CS1 showed a significantly higher (P < 0.001) selectivity in lignin degradation compared to CS2. Both strains showed peak activity of laccase and manganese peroxidase (MnP) at week 1. CS1 showed a significantly higher (P < 0.001) laccase activity, but lower (P = 0.008) MnP activity compared to CS2. Conclusion Both CS strains improved the nutritive value of wheat straw. Variation between strains was clearly demonstrated by their growth pattern and enzyme activities. Significance and Impact of Study The differences among the two strains provide an opportunity for future selection and breeding programs in improving the extent and selectivity of lignin degradation in agricultural biomass. This article is protected by copyright. All rights reserved.

Lignin degradation by white-rot fungi is an essential step in terrestrial carbon cycling and has great potential for biotechnological applications. Selective white-rot fungi have been recognized for their ability to effectively delignify lignocellulose, but the underlying mechanisms, particularly in situ, have largely remained elusive to date. In this work, we elucidate specific degradation routes of β-O-4 aryl ethers in actual lignocellulosic biomass for the industrially relevant selective white-rot fungus Ceriporiopsis subvermispora. Multidimensional NMR and py-GC-MS analyses together with enzymatically synthesized model compounds enabled, for the first time, the identification of various diagnostic lignin cleavage products in residual wheat straw. Our results support that in situ ligninolysis by C. subvermispora is initiated by single-electron transfer, which then cascades into the cleavage of Cα-Cβ, Cβ-O, and O-4-aryl bonds of β-O-4 aryl ethers. The high abundance of 1-(benzyl)-2,3-dihydroxypropan-1-ones indicated that β-O-|4 cleavage is a more important pathway than previously considered. Our approach highlights key diagnostic substructures for providing mechanistic insight into fungal ligninolysis.

Miscanthus was pretreated with the fungus Ceriporiopsis subvermispora under non-sterile conditions, using sterile miscanthus that had been previously colonized with the fungus as the inoculum. Inoculum ratios equal to or greater than 30% yielded a successful pretreatment, enhancing the enzymatic digestibility of miscanthus by 3- to 4-fold over that of raw miscanthus, which was comparable with the fungal pretreatment under sterile conditions. This enhanced digestibility was linearly correlated with lignin degradation. Although cellulose loss of up to 13% was observed for the successful non-sterile pretreatments, the final glucose yield was 3-4 times higher than that of raw miscanthus and comparable to that of the sterile pretreated miscanthus. A time course study showed that maximum glucose yield can be achieved with a pretreatment time of 21 days.

This study evaluated the biodepolymerization of biochar from cotton stalks gasification using four fungal strains Phanerochaete chrysosporium, Ceriporiopsis subvermispora, Postia placenta, and Bjerkandera adusta. After the biological treatment, the liquid and solid phases were analyzed using UV–Vis and FT-IR spectroscopy. The biochar’s largest depolymerization was obtained by C. subvermispora. The depolymerization kinetic of C. subvermispora showed the production of laccase and manganese peroxidase and a correlation between the depolymerization and the production of ligninolytic enzymes. The highest production of ligninolytic enzymes and depolymerization was observed after 24 days. The modifications in the UV–Vis and FT-IR spectra exhibited the presence of functional groups related with humic substances from cultures with C. subvermispora and cotton stalks biochar.

A rapid and complete colonization of lignocellulosic biomass by white-rot fungi is a prerequisite for an efficient and contamination free delignification. To improve the colonization, mycelium growth and delignification by Ceriporiopsis subvermispora and Lentinula edodes of wheat straw, with addition of 2, 4, 6 and 10% of wheat bran for 0, 2, 4, 8 and 12 weeks, was evaluated. The ergosterol content, chemical composition and in vitro gas production (IVGP) by ruminal microorganisms were determined. The fungal biomass and starch amount was significantly (P < 0.001) affected by the inclusion level of wheat bran and incubation time. An increase in added wheat bran increased the fungal biomass with different effects observed for the two fungi. The added starch from the wheat bran was degraded by C. subvermispora, mainly during the first two weeks (up to 44%), whereas, L. edodes degraded starch during 12 weeks up to 83%. The supplementation of wheat bran, however, did not affect (P > 0.05) the amount of hemicellulose and lignin at the end of the incubation. There was no significant effect of wheat bran addition on the IVGP for the two fungal treatments. Supplementation of wheat straw with wheat bran resulted in a quicker fungal colonization, which is useful for C. subvermispora, a fungus that forms only thin mycelium and is more prone to contamination than L. edodes.

The basidiomycete Ceriporiopsis subvermispora is a selective white-rot fungus that mainly depolymerizes lignin and hemicellulose leaving cellulose almost intact. Since it produces hydrolytic enzymes, it has potential to grow on different lignocellulosic substrates in solid-state conditions and to transform lignocellulose into a variety of valuable phenolic compounds. In this work, grape pomace, a waste that remains after winemaking process, was biologically treated with C. subvermispora during fifteen days. Nineteen different individual phenolic compounds were analysed by gradient UHPLC method. All analyses were conducted before, during and after the biological treatment. The biological treatment of grape pomace had positive effect on liberation of phenolic compounds from its complex lignocellulosic structure. The highest content of phenolic acids such as p-hydroxybenzoic (0.01 mg/gdb), protocatechuic (0.20 mg/gdb), p coumaric (0.01 mg/gdb), syringic (0.14 mg/gdb), caffeic (0.01 mg/gdb), ferulic (0.01 mg/gdb), and vanillic (0.05 mg/gdb) acid was observed after one day and for gallic acid (0.28 mg/gdb) after two day of biological treatment.

Background The white-rot fungi Ceriporiopsis subvermispora (Cs), Pleurotus eryngii (Pe), and Lentinula edodes (Le) have been shown to be high-potential species for selective delignification of plant biomass. This delignification improves polysaccharide degradability, which currently limits the efficient lignocellulose conversion into biochemicals, biofuels, and animal feed. Since selectivity and time efficiency of fungal delignification still need optimization, detailed understanding of the underlying mechanisms at molecular level is required. The recently developed methodologies for lignin quantification and characterization now allow for the in-depth mapping of fungal modification and degradation of lignin and, thereby, enable resolving underlying mechanisms. Results Wheat straw treated by two strains of Cs (Cs1 and Cs12), Pe (Pe3 and Pe6) and Le (Le8 and Le10) was characterized using semi-quantitative py-GC–MS during fungal growth (1, 3, and 7 weeks). The remaining lignin after 7 weeks was quantified and characterized using 13C lignin internal standard based py-GC–MS and whole cell wall HSQC NMR. Strains of the same species showed similar patterns of lignin removal and degradation. Cs and Le outperformed Pe in terms of extent and selectivity of delignification (Cs ≥ Le >> Pe). The highest lignin removal [66% (w/w); Cs1] was obtained after 7 weeks, without extensive carbohydrate degradation (factor 3 increased carbohydrate-to-lignin ratio). Furthermore, though after treatment with Cs and Le comparable amounts of lignin remained, the structure of the residual lignin vastly differed. For example, Cα-oxidized substructures accumulated in Cs treated lignin up to 24% of the total aromatic lignin, a factor two higher than in Le-treated lignin. Contrarily, ferulic acid substructures were preferentially targeted by Le (and Pe). Interestingly, Pe-spent lignin was specifically depleted of tricin (40% reduction). The overall subunit composition (H:G:S) was not affected by fungal treatment. Conclusions Cs and Le are both able to effectively and selectively delignify wheat straw, though the underlying mechanisms are fundamentally different. We are the first to identify that Cs degrades the major β-O-4 ether linkage in grass lignin mainly via Cβ–O–aryl cleavage, while Cα–Cβ cleavage of inter-unit linkages predominated for Le. Our research provides a new insight on how fungi degrade lignin, which contributes to further optimizing the biological upgrading of lignocellulose.

International audience; Fungal pretreatment by Ceriporiopsis subvermispora of two forest residues (hazel and acacia branches) and two agricultural lignocellulosic residues (barley straw and sugarcane bagasse) were studied as a pretreatment to improve their subsequent anaerobic digestion for methane production. Biomass samples were grinded to 2 ranges of particle sizes (

Efficient lignin depolymerization is unique to the wood decay basidiomycetes, collectively referred to as white rot fungi. Phanerochaete chrysosporium simultaneously degrades lignin and cellulose, whereas the closely related species, Ceriporiopsis subvermispora, also depolymerizes lignin but may do so with relatively little cellulose degradation. To investigate the basis for selective ligninolysis, we conducted comparative genome analysis of C. subvermispora and P. chrysosporium . Genes encoding manganese peroxidase numbered 13 and five in C. subvermispora and P. chrysosporium , respectively. In addition, the C. subvermispora genome contains at least seven genes predicted to encode laccases, whereas the P. chrysosporium genome contains none. We also observed expansion of the number of C. subvermispora desaturase-encoding genes putatively involved in lipid metabolism. Microarray-based transcriptome analysis showed substantial up-regulation of several desaturase and MnP genes in wood-containing medium. MS identified MnP proteins in C. subvermispora culture filtrates, but none in P. chrysosporium cultures. These results support the importance of MnP and a lignin degradation mechanism whereby cleavage of the dominant nonphenolic structures is mediated by lipid peroxidation products. Two C. subvermispora genes were predicted to encode peroxidases structurally similar to P. chrysosporium lignin peroxidase and, following heterologous expression in Escherichia coli , the enzymes were shown to oxidize high redox potential substrates, but not Mn 2+ . Apart from oxidative lignin degradation, we also examined cellulolytic and hemicellulolytic systems in both fungi. In summary, the C. subvermispora genetic inventory and expression patterns exhibit increased oxidoreductase potential and diminished cellulolytic capability relative to P. chrysosporium .

The variable C-terminal tail of manganese peroxidases, a group of enzymes involved in lignin degradation, is implicated in their catalytic and stability properties, as shown by new crystal structures, molecular-simulation and directed-mutagenesis data. Based on this structural–functional evaluation, short and long/extralong manganese peroxidase subfamilies have been accepted; the latter are characterized by exceptional stability, while it is shown for the first time that the former are able to oxidize other substrates at the same site where manganese(II) is oxidized., The genome of Ceriporiopsis subvermispora includes 13 manganese peroxidase (MnP) genes representative of the three subfamilies described in ligninolytic fungi, which share an Mn2+-oxidation site and have varying lengths of the C-terminal tail. Short, long and extralong MnPs were heterologously expressed and biochemically characterized, and the first structure of an extralong MnP was solved. Its C-terminal tail surrounds the haem-propionate access channel, contributing to Mn2+ oxidation by the internal propionate, but prevents the oxidation of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS), which is only oxidized by short MnPs and by shortened-tail variants from site-directed mutagenesis. The tail, which is anchored by numerous contacts, not only affects the catalytic properties of long/extralong MnPs but is also associated with their high acidic stability. Cd2+ binds at the Mn2+-oxidation site and competitively inhibits oxidation of both Mn2+ and ABTS. Moreover, mutations blocking the haem-propionate channel prevent substrate oxidation. This agrees with molecular simulations that position ABTS at an electron-transfer distance from the haem propionates of an in silico shortened-tail form, while it cannot reach this position in the extralong MnP crystal structure. Only small differences exist between the long and the extralong MnPs, which do not justify their classification as two different subfamilies, but they significantly differ from the short MnPs, with the presence/absence of the C-terminal tail extension being implicated in these differences.

The aim of this study was to set parameters of repeated-batch cultivation of Ceriporiopsis subvermispora for laccase production and evaluate the efficiency of this type of cultivation for production of selected enzyme. The suitable conditions for repeated-batch cultivation were designed on the base of study of batch cultivation of white-rot fungus C. subvermispora. C. subvermispora was cultivated in media with different concentration of casein hydrolysate as nitrogen source and glucose as carbon source. A suitable concentration of casein hydrolysate to stimulate the laccase production was 1.5 and 2.5 g/L. Laccase production was started at certain critical concentration of glucose (5 g/L). In order to improve laccase production by repeated-batch cultivation of C. subvermispora, glucose was tested in concentration 10 g/L and casein hydrolysate in concentration 1.5 g/L. During a repeated-batch cultivation was measured increase laccase activities from 177.8 to 266 U/L. It was also observed, the cultivation time needed to reach maximum laccase production was shortened to 10 days.

The present work aimed to study sugarcane bagasse fibers pre-treated with fungi and using NaOH/ anthraquinone (AQ) in chemical pulping processes for applications in composite materials. Bagasse was decayed with 250 mg of Ceriporiopsis subvermispora inoculum in a 20 L bioreactor. After that, samples were submitted to similar conditions of decaying without inoculum charge. Decayed and undecayed fibers were treated with NaOH 12.5 wt%, 0.15 of AQ and a 12:1 (v/w) liquor:bagasse ratio at 160°C. Then, all obtained fibers were characterized according to their chemical composition. Dried biotreated (decayed) and control (undecayed) fibers were mixed through an extruder process with polypropylene. Later, composite granulates were injected directly in mold with cavities for tensile, flexural and shear tests. Composite materials with 10 and 20 wt% fibers were submitted to static mechanical standard tests and DMA (Dynamic Mechanical Analysis) to evaluate the effect of biotreatment. Biotreatment, cook time (pulping), and fiber content contributed to improvements in the mechanical properties of the composites. The interface between fiber and matrix was increased with the biotreatment and pulping of fibers. Furthermore, DMA results also showed that fiber incorporation into PP improved the modulus, mainly for biotreated fibers/PP composites. The Tg (tan δ data) from composites was dislocated at lower temperatures with respect to neat PP due to the influence of fibers on matrix.

This work evaluates linoleic acid peroxidation reactions initiated by Fe 3+ -reducing compounds recovered from Eucalyptus grandis , biotreated with the biopulping fungus Ceriporiopsis subvermispora . The aqueous extracts from biotreated wood had the ability to reduce Fe 3+ ions from freshly prepared solutions. The compounds responsible for the Fe 3+ -reducing activity corresponded to UV-absorbing substances with apparent molar masses from 3 kDa to 5 kDa. Linoleic acid peroxidation reactions conducted in the presence of Fe 3+ ions and the Fe 3+ -reducing compounds showed that the rate of O 2 consumption during peroxidation was proportional to the Fe 3+ -reducing activity present in each extract obtained from biotreated wood. This peroxidation reaction was coupled with in-vitro treatment of ball-milled E. grandis wood. Ultraviolet data showed that the reaction system released lignin fragments from the milled wood. Size exclusion chromatography data indicated that the solubilized material contained a minor fraction representing high-molar-mass molecules excluded by the column and a main low-molar-mass peak. Overall evaluation of the data suggested that the Fe 3+ -reducing compounds formed during wood biodegradation by C. subvermispora can mediate lignin degradation through linoleic acid peroxidation.

In the present study, it was evaluated how two different culture conditions for the biotreatment of Eucalyptus grandis by Ceriporiopsis subvermispora affect a subsequent high-yield kraft pulping process. Under the varied culture conditions investigated, different extracellular enzyme activities were observed. Manganese-peroxidase (MnP) secretion was 3.7 times higher in cultures supplemented with glucose plus corn-steep liquor (glucose/CSL) as compared to non-supplemented (NS) cultures. The biotreated samples underwent diverse levels of wood component degradation as losses of weight and lignin were increased in glucose/CSL cultures. Mass balances for lignin removal during kraft pulping showed that delignification was facilitated when both biotreated wood samples were cooked. Delignification efficiency did not correlate positively with MnP levels in the cultures. On the other hand, biopulps from NS and glucose/CSL cultures saved 27% and 38% beating time to achieve 28° Schopper-Riegler freeness during refining, respectively. Biopulps disposed of decreased tensile and tear resistances, thus easier refining of the biokraft pulps seems to be a consequence of less resistant fiber walls. Improved beatability of biopulps was tentatively related to short fibers and fines formation during refining. We suggest that to some extent polysaccharide depolymerization occurred during the biotreatment, which also resulted in diminished pulp yields in the case of glucose/CSL cultures.

The effect of different culture conditions have been evaluated concerning the extracellular enzyme activities of the white-rot fungus Ceriporiopsis subvermispora growing on Eucalyptus grandis wood. The consequence of the varied fungal pretreatment on a subsequent chemithermomechanical pulping (CTMP) was addressed. In all cultures, manganese peroxidase (MnP) and xylanase were the predominant extracellular enzymes. The biopulping efficiency was evaluated based on the amount of fiber bundles obtained after the first fiberizing step and the fibrillation levels of refined pulps. It was found that the MnP levels in the cultures correlated positively with the biopulping benefits. On the other hand, xylanase and total oxalate levels did not vary significantly. Accordingly, it was not possible to determine whether MnP accomplishes the effect alone or depends on synergic action of other extracellular agents. Pulp strength and fiber size distribution were also evaluated. The average fiber length of CTMP pulps prepared from untreated wood chips was 623 μm. Analogous values were observed for most of the biopulps; however, significant amounts of shorter fibers were found in the biopulp prepared from wood chips biotreated in cultures supplemented with glucose plus corn-steep liquor. Despite evidence of reduced average fiber length, biopulps prepared from these wood chips presented the highest improvement in tensile indexes (+28% at 23° Schopper-Riegler).

In biopulping, efficient wood colonization by a selected white-rot fungus depends on previous wood chip decontamination to avoid the growth of primary molds. Although simple to perform in the laboratory, in large-scale biopulping trials, complete wood decontamination is difficult to achieve. Furthermore, the use of fungal growth promoters such as corn steep liquor enhances the risk of culture contamination. This paper evaluates the ability of the biopulping fungus Ceriporiopsis subvermispora to compete with indigenous fungi in cultures of fresh or poorly decontaminated Eucalyptus grandis wood chips. While cultures containing autoclaved wood chips were completely free of contaminants, primary molds grew rapidly when non-autoclaved wood chips were used, resulting in heavily contaminated cultures, regardless of the C. subvermispora inoculum/wood ratio evaluated (5, 50 and 3000 mg mycelium kg−1 wood). Studies on benomyl-amended medium suggested that the fungi involved competed by consumption of the easily available nutrient sources, with C. subvermispora less successful than the contaminant fungi. The use of acid-washed wood chips decreased the level of such contaminant fungi, but production of manganese peroxidase and xylanases was also decreased under these conditions. Nevertheless, chemithermomechanical pulping of acid-washed samples biotreated under non-aseptic conditions gave similar fibrillation improvements compared to samples subjected to the standard biodegradation process using autoclaved wood chips.

The herbicide diuron (3-(3,4-dichlorophenyl)-1,1-dimethylurea) is used in many agricultural crops and non-crop areas worldwide, leading to the pollution of the aquatic environment by soil leaching. White rot fungi and its lignin modifying enzymes, peroxidases and laccases, are responsible for its degradation. Therefore, it is of interest to explore the potential use of Ceriporiopsis subvermispora laccase (CersuLac1) in the biotransformation of this herbicide by using its enzyme laccase. However, the structure of laccase from Ceriporiopsis subvermispora is still unknown. Hence, a model of laccase was constructed using homology modeling. The model was further used to dock p-methylbenzoate in the presence of four copper ions to analyze molecular basis of its binding and interaction. The ligand-protein interaction is stereo-chemically favorable in nature. The presence of the single protonated Lys457 was necessary for catalysis, being coordinated by a cupper ion. The best pose of diuron on CersuLac1 has a theoretical Ki of 2.91 mM. This is comparable to the KM values for laccases from other organisms with similar compounds. Thus, we document the insights for the potential use of laccase from Ceriporiopsis subvermispora in the biotransfrormation of diuron.

Loblolly pine (Pinus taeda) chips treated with Ceriporiopsis subvermispora for two or four weeks were pulped with different combinations of kraft pulping conditions to obtain a better understanding of the interaction between the fungal action and the pulping variables. Two different levels of effective alkali (18 or 22%), two times at maximum Tmax (60 or 90 min), 22% sulfidity, and a Tmax of 170°C, were used. The best delignification without adversely affecting pulp viscosity was found in pulps made from chips treated with the fungus for 2 weeks and at the mildest pulping conditions. At all pulping conditions there was a substantial decrease in the amount of rejects with 2 weeks of fungal treatment. Pulps from fungally‐treated chips refined more easily than the control pulp and strength properties of pulps of fungally‐treated chips were superior to those of the control pulp.

The effect of treating loblolly pine (Pinus taeda) chips with the fungus Ceriporiopsis subvermispora was investigated by assessing the wood changes at 2 and 4 weeks incubation relative to a control (no fungal treatment). Scanning electron microscopy indicated that during the first 2 weeks of colonization, C. subvermispora had grown over the surface of the chips and also within ray cells. A mass loss of 5% occurred after 2 weeks, 6% after four weeks. The extractives were reduced 23% in the first 2 weeks of fungal treatment and 32% after 4 weeks. Lignin loss was statistically insignificant at 2 weeks but reached 8% of the original lignin after 4 weeks. The carbohydrate content of the treated wood showed no significant differences from the control after 2 weeks and only minor losses after 4 weeks. The fraction of the wood soluble in 1% NaOH was 14.9% at 2 weeks compared to 11.5% for the control chips and increased to 18.8% after 4 weeks. This indicated only a mild decay due to the fungal treatment. ...

The capabilities of four white-rot fungi to improve the digestibility of sugarcane bagasse for ruminants were determined. Bagasse was cultured with Lentinula edodes, Pleurotus eryngii, Pleurotus salmoneostramineus, Ceriporiopsis subvermispora (ATCC 90467) or C. subvermispora (CZ-3) at 26°C for 8, 12 or 16 weeks. The in vitro organic matter digestibility (IVOMD) and in vitro neutral detergent fiber digestibility (IVNDFD) in untreated bagasse were 45.6 and 40.3%, respectively. Meanwhile, the bagasse cultured with L. edodes and two strains of C. subvermispora, respectively, for 12 weeks, were elevated to 68.6 and 59.1%, 60.6 and 49.9% and 59.9 and 49.0%, respectively. In contrast, the IVOMD and the IVNDFD in bagasse cultured with P. eryngii and P. salmoneostramineus were the same or lower than those in untreated bagasse. In vitro gas production (IVGP) in bagasse cultured with L. edodes and two strains of C. subvermispora for 12 weeks was also higher than that of untreated bagasse. These changes in IVOMD, IVNDFD and IVGP demonstrate that L. edodes has a higher capability of improving the digestibility for ruminants than C. subvermispora, P. eryngii or P. salmoneostramineus.

Production of cellulases and hemicellulases by Ceriporiopsis subvermispora cultured on wood chips of Eucalyptus grandis and Pinus taeda was studied. Xylanase, endo-1,4 β-glucanase, mannanase, β-glucosidase and β-xylosidase were detected in enzymatic extracts obtained after 30 days of cultivation. P. taeda gave rise to higher enzyme production compared to E. grandis . Biochemical characteristics of cellulases and hemicellulases produced in both wood species were almost the same. The optimum pH for the enzymes was between 4.0 and 5.0 and the optimum temperature was 60 °C. Thermal stability of the enzymes at pH 5.0 was appreciable up to 50 °C. The profile of proteins contained in extracts recovered from both biotreated wood species showed a similar number of bands during SDS-PAGE electrophoresis. Five bands with molecular weight ranging from 50 to 85 kDa and two bands with high molecular weight (100–200 kDa) were observed. Activity staining for xylanases indicated two enzymes with molecular weight of 80 and 33 kDa.

Near‐infrared (NIR) spectroscopy was evaluated as an analytical tool for monitoring changes in Pinus taeda (loblolly pine) induced by the white‐rot fungus Ceriporiopsis subvermispora. Intensities of several NIR bands increased with biodegradation time, but a direct correlation between band intensities and biodegradation periods or weight losses due to the biotreatment were not observed. On the other hand, the intensities of NIR bands correlated with properties reflecting the macromolecular characteristics of the components of the biotreated wood samples. Bio‐kraft pulps were also characterized by wet chemical analysis and NIR spectroscopy. In this case, appropriate prediction models related NIR spectral information with the chemical composition of the pulps, r 2 values ranged from 0.96 to 0.99.

The wood colonising white-rot basidiomycete Ceriporiopsis subvermispora is able to degrade lignin in preference to cellulose. To differentiate between fungal strains and to estimate their delignification behaviour, both in an early stage of degradation and over a specific period, is important for the wood industry. Mid infrared (MIR) and near infrared (NIR) spectra were taken from 60 milled spruce wood samples and their total lignin content was determined by wet laboratory methods. Good correlations were found between the MIR band–height ratio (H1510 cm−1 / H897 cm−1) and the lignin content ( r = 0.965) and between the NIR band height at 5978 cm−1 (1673 nm) taken from spectra in the second derivative mode and the lignin content ( r = 0.956). Furthermore, good linear correlations between the band–height ratios calculated from the MIR spectra and the amplitudes of the band around 5978 cm−1 (1673 nm) of NIR spectra in the second derivative mode were found for the calibration samples ( r = 0.934) and for the fungal-treated samples ( r = 0.984). The good correlation found between the MIR band–height ratio and the band height from NIR spectra in the second derivative mode could be interesting if calibrations exist for MIR (or NIR) to predict samples measured in the NIR (or MIR). MIR and NIR spectra recorded from milled spruce wood shavings that had been subjected to fungal treatment with three strains of Ceriporiopsis subvermispora (CBS 347.63, FPL 90.031 and FPL 105.752), for a period of up to 14 days, were investigated to see if these spectroscopic techniques could replace chemical methods. It is shown that the relative degree of delignification can be obtained directly from NIR spectra in the second derivative mode measuring the amplitude of a distinct band and from MIR spectra normalised with respect to the band at 897 cm−1. Subjecting the spectra to principal component analysis (PCA) made it possible to study the time course along a PC axis. The use of an appropriate NIR wavenumber range subjected to PCA led to a scores plot that made it possible to differentiate between the three strains of C. subvermispora along one axis. It was also possible to give a time course and an indication of the relative degree of delignification along the second axis. In both cases, 99% of the data variance was explained with the first two PCs. A similar time course was obtained from MIR spectra, but the strains could not be separated well. Besides strain differentiation and examination of delignification, some practical applications (for example, in the pulp and paper industries, fungi-screening, evaluation of wood preservatives) are discussed. The results clearly demonstrate that it is possible to compare and differentiate between the strains without applying time-consuming chemical methods. The examination of NIR spectra is sufficient.

Scanning UV-microspectrophotometry was used to investigate the topochemistry of lignin removal from pine wood (Pinus taeda and P. elliottii) chips during biopulping involving wood treatment with Ceriporiopsis subvermispora (Pilat) Gilbn. & Ryv. followed by alkaline sulfite/anthraquinone delignification. A delignification front starting from the lumen towards the compound middle lamella was clearly observed in micrographs recorded from individual cell wall layers of wood samples biotreated for 30 days. Lignin was removed without cell wall erosion. UV-micrographs of wood samples cooked for a short time (90 min pulping) showed that the S2 of biotreated samples are more homogeneously delignified compared to the S2 of the undecayed controls. Similarly, the compound middle lamella and cell corners are also more delignified in biotreated samples. On the other hand, UV-micrographs of samples cooked for a long time (150 min pulping at 170°C) showed that there are no significant differences in the contents of residual lignin retained in the S2 of undecayed and biotreated wood samples.

Pine wood chips were treated for 30 days with Cemporiopsis subvermispora in 20 dm3 bioreactors. A typical selective biodelignification was observed. The biotreated wood chips and undecayed controls were subjected to modified alkaline sulfite/anthraquinone (ASA) cooking at 170 °C or 175 °C applying varying cooking times ranging from 30 to 270 min. In all cases, the residual lignin content of the pulps prepared from biologically pretreated wood chips was lower than that of the control pulp. With increasing cooking time, however, the differences in kappa number became smaller. Wood chips cooked for a short time required mechanical refining for fiber liberation. A disk-refining step resulted in pulps with low reject content (0.4%) and high screened yield (56–60%). In this case, the use of biotreated wood chips provided pulps with significantly lower kappa numbers than for the control pulp (71 and 83, respectively). The pulp from biologically pretreated wood fibrillated rapidly, reaching 20° SR in only 38 min beating time in a Jokro mill, while the control pulp required 56 min to reach the same beating degree. Although easier to beat, the biopretreated pulps showed tensile and burst indices similar to those of the control samples. However, their tear indices were always lower. Easier delignification after wood biotreatment was not observed for the reactions performed at long cooking times. Oxygen delignification of biotreated and conventional ASA pulps with low kappa numbers reduced kappa number and improved brightness considerably with the biotreated pulps being favored by a better preservation of viscosity. Copyright © 2004 Society of Chemical Industry

Summary The effect of biological pretreatment of sisal with several white rot fungi on the energy consumption in refining and on the mechanical properties of the pulps was evaluated in this investigation. Improvements were realized in all the mechanical properties (22–66 %) and a reduction in the energy consumption of > 39% was realized for the treated pulps with the different fungi. The best strength improvement and energy reduction results overall were obtained with the white-rot fungus, Ceriporiopsis subvermispora. The incubation time was optimized for this fungus with the optimum mechanical properties obtained with a two week treatment time.

A novel series of long-chain unsaturated dicarboxylic acids consisting of a long aliphatic chain attached to the C-3 position of itaconic acid has been identified by gas chromatography/mass spectrometry during in vitro decay of eucalypt wood by the white-rot basidiomycete Ceriporiopsis subvermispora. The major compounds were identified as tetradecyl-, 7-hexadecenyl- and hexadecylitaconic acids by their mass fragmentation patterns. Other members of the same compound series, identified as dodecanyl-, tridecanyl-, tetradecenyl-, pentadecanyl-, octadecenyl- and octadecanylitaconic acids, were present in very minor amounts or traces. Whereas hexadecenylitaconic acid has already been reported in cultures of C. subvermispora, to our knowledge this is the first report of the presence of the other alkylitaconic acids in fungal cultures. These new alkylitaconic-type metabolites may constitute a source for peroxidizable lipids involved in lignin degradation during wood decay by C. subvermispora and other white-rot basidiomycetes.

Biosulfite pulping of Eucalyptus grandis wood chips pretreated with ten strains of selected white-rot fungi was investigated. The biopulping efficiency was evaluated based on fungal pretreatment time (5, 7 and 10 days) and properties of biosulfite pulp (pulp yield, kappa number, brightness, alkali solubility S10 and S18, and viscosity) in comparison to the controls. Pretreatment of eucalyptus wood chips with Ceriporiopsis subvermispora SS-3 in the absence of an additional carbon source for 10 days resulted in a substantial decrease in kappa number (by 29%) and increase in brightness (by 12%). On the other hand 5% of the pulp yield was lost, and both viscosity and alkali solubility (S10 and S18) were retained at levels comparable to the control. When the fungal incubation time was reduced to 5 days, a 5% reduction of kappa number, 11% increase in brightness and no loss of pulp yield was observed. C. subvermispora SS-3 was therefore selected for further optimization studies of biosulfite pulping for dissolving pulp-directed manufacture

This study deals with pretreatment of wheat straw with lignin-degrading fungi and its effect on chemical pulping. Ceriporiopsis subvermispora strains, which preferentially attack the lignin, were used for biochemical pulping of bagasse. Treatment of depithed bagasse with different strains of C. subvermispora reduced the kappa number by 10-15% and increased unbleached pulp brightness by 1.1-2.0 ISO points on chemical pulping at the same alkali charge. Bleaching of biopulps at the same chemical charge increased final brightness by 4.7-5.6 ISO points and whiteness by 10.2-11.4 ISO points. Fungal treatment did not result in any adverse effect on the strength properties of pulp.

New ceriporic acids—alkadienyl and alkenyl itaconic acids having a bis-allyl (3-[(Z,Z)-hexadec-7,10-dienyl]-itaconic acid; ceriporic acid G) and a monoene (3-[(Z)-octadec-9-enyl]-itaconic acid; ceriporic acid H) structure in their side chains—were isolated from the cultures of the selective lignin-degrading fungus Ceriporiopsis subvermispora. The new metabolites ceriporic acid G and H were synthesized by a cross-aldol condensation and a Grignard substitution reaction, respectively. Ceriporic acid G triggered the manganese peroxidase (MnP)-catalyzed lipid peroxidation and decomposed a recalcitrant non-phenolic lignin substructure model compound. Except for simple fatty acids, this is the first report of a fungal metabolite that induced ligninolytic lipid peroxidation.

Spruce wood that had been degraded by white-rot fungi (Trametes versicolororCeriporiopsis subvermispora)and suffered mass losses up to 17% was investigated by transmission Fourier transform infrared (FT-IR) imaging microscopy. A significant marker during incipient simultaneous white-rot (T. versicolor) was the cleavage of glycosidic bonds of polysaccharides that preceded their metabolisation. Simultaneous white-rot processes were also characterised by a relative decrease of the overall lignin content and a relative accumulation of wood polysaccharides. No early marker was found for selective white-rot (C. subvermispora) that removes mainly lignin by an oxidative process. This feature was detected only in wood samples exhibiting mass losses higher than 12%. Furthermore, it was shown, that simultaneous and selective white-rot processes were unevenly distributed within the wood samples but quite evenly distributed within single tracheids.

European black pine (Pinus nigra Am.) chips were treated with the white-rot fungus Ceriporiopsis subvermispora for periods ranging from 20 to 100 days. The effects of pretreatment on the chemical composition of wood and kraft pulping were investigated. The results showed that fungal pretreatment reduced the lignin and extractive content of wood chips. Also, weight losses occurred. Kappa number, viscosity, and reject ratio of biokraft pulps decreased. Biokraft pulps gave better response to beating, which led to significant energy saving during refining. The tear index, burst index, and tensile index of biokraft pulps were found to be lower than those of kraft pulps. However, the tensile index and burst index of 20-day biotreated and unbeaten pulp was higher than those of kraft pulp. Also, the tear index of 20-day biotreated and beaten pulp was higher than that of kraft pulp. The brightness of biokraft pulps decreased irregularly with increasing incubation time. (C) 2011 Elsevier Ltd. All rights reserved.

Paper optical properties handsheets, whereas brightness and light scattering coefficient of the handsheets were decreased after all of the fungal treatments. Opacity of the handsheets after the fungal treatments remained unchanged. Loblolly pine (Pinus taeda) White-rot fungi Based on energy savings and improvements in the strength properties, regardless of bioreactor type, the white-rot fungus Ceriporiopsis subvermispora appeared to be superior to the other white-rot fungi tested. When incubated in stationary tray bioreactors, C. subvermispora caused only 6% weight loss, saved 42% energy during fiberization and refining, improved burst index by 32% and tear index by 67%, as compared to the control.

Ceriporiopsis subvermispora é um fungo filamentoso, muito seletivo na degradação de lignina e, por isso, tem sido uma das espécies mais estudadas no processo de biopolpação. A biopolpação consiste em um tratamento biológico da madeira que antecede etapas convencionais de polpação, proporcionando níveis de economia de energia elétrica no processo que podem atingir valores de 30 a 40%. Para degradar a lignina, esse fungo secreta a enzima manganês-peroxidase (MnP), a qual requer um ácido carboxílico para quelar e transportar íons Mn3+ oriundos do seu ciclo catalítico. O complexo quelante-Mn3+ degrada apenas frações fenólicas da lignina, porém pode também iniciar a peroxidação de lipídeos e com isso gerar radicais peroxila que apresentam capacidade oxidativa suficiente para degradar estruturas não-fenólicas da lignina. Com base nesses aspectos, o presente trabalho teve o objetivo de avaliar a degradação de lignina por reações que envolvem a peroxidação de ácido linoléico iniciadas por metabólitos extracelulares extraídos de cultivos de C. subvermispora. Também foram avaliados sistemas miméticos baseados nos íons Fe2+ e Mn3+ como iniciadores das mesmas reações. Essencialmente, foi estudada a degradação de lignina in vitro em reações iniciadas por sistemas compostos que incluíram, MnP/Mn+2/H2O2, Fe3+/agentes redutores de Fe3+ produzidos durante a biodegradação da madeira, íons Mn+3 ou íons Fe+2, todos adicionados ao ácido linoléico. Para realizar esse estudo foi necessário preparar, tanto MnP, quanto compostos redutores de Fe3+, em cultivos de C. subvermispora. Também foram preparados e caracterizados dois substratos para as reações em estudo. Esses substratos compreenderam um complexo lignina-carboidrato (CLC) e um modelo de um material lignocelulósico completo, porém moído e livrado de toda a fração de extrativos. Nos dois casos, o material de partida foi à madeira de Eucalyptus grandis. A caracterização química desses substratos indicou um teor de lignina de 44,8% e 29,0%, respectivamente. Reações de peroxidação de ácido linoléico iniciadas pelos sistemas em estudo mostraram que todos foram efetivos para esse propósito, sendo que as maiores taxas de consumo de oxigênio durante essas reações foram observadas nos meios reacionais que continham Fe2+ em solução. O CLC inibiu as reações de peroxidação quando adicionado ao meio reacional em concentraçãoes maiores do que 0,3 mg/mL. Entretanto, reações prolongadas por 72 h com o CLC numa concentração inicial de 1 mg/mL indicaram que ele sofreu despolimerização. As vias de degradação da lignina contida no CLC ou na madeira de E. grandis moída envolveram a despolimerização da molécula, ou simplesmente a oxidação das cadeias laterais e das estruturas fenólicas livres. Quando o sistema foi baseado na ação de MnP/Mn+2/H2O2/ácido linoléico, foram comprovadas vias de degradação da lignina que envolveram desde a simples oxidação do carbono-α até a quebra de ligações do tipo β-O-4 e/ou entre os carbonos α e β. Os resultados obtidos corroboraram dados anteriormente publicados para a ação de C. subvermispora in vivo. Uma exceção foi à observação da reação de simples oxidação do Cα nos sistemas in vitro, que havia sido descartada em trabalhos anteriores que se basearam na caracterização de lignina contida em madeira biotratada por C. subvermispora (sistema in vivo). Os resultados permitiram concluir que vários sistemas miméticos podem iniciar a peroxidação de ácido linoléico in vitro. Quando essas reações foram conduzidas na presença de lignina (CLC ou E. grandis moído) foi possível observar transformações importantes na estrutura da lignina que eventualmente poderiam ser exploradas, por exemplo, em etapas de processos de branqueamento de polpas kraft. The white-rot fungus Ceriporiopsis subvermispora degrades lignin selectively, being one of the most studied species in biopulping. Biopulping consists of a biological treatment of wood that precedes conventional pulping stages. The process can provide up to 30-40% of energy savings in mechanical pulping. To degrade lignin, this fungus secretes the enzyme manganese-peroxidase (MnP), which needs carboxylic acids to chelate and transport Mn3+ ions formed in the catalytic cycle of the enzyme. The chelate-Mn3+ complex is able to degrade phenolic structures of lignin; however, can also initiate lipid peroxidation reactions generating peroxyl radicals that are able to degrade nonphenolic lignin structures. Based on this background, the aim of this work was to evaluate lignin degradation through linoleic acid peroxidation reactions initiated by extracellular metabolites recovered from C. subvermispora cultures. Some biomimetic systems based on Fe2+ and Mn3+ ions were also evaluated as initiators of such reactions. The lignin degradation was studied in reaction systems composed of MnP/Mn+2/H2O2, Fe3+-reducing compounds produced during wood biodegradation by C. subvermispora, Mn+3 or Fe+2 ions, all of them in the presence of linoleic acid. To perform this study, MnP and Fe3+-reducing compounds were initially produced in C. subvermispora cultures. Two different reaction substrates were also prepared. One was a lignin-carbohydrate complex (LCC) and, the other, was a complete lignocellulosic material that was milled and extracted to remove the extractive fraction. Both substrates were prepared from Eucalyptus grandis wood. The chemical characterization of the substrates showed 44.8 % and 29.0 % of total lignin, respectively. Linoleic acid peroxidation reactions initiated by the studied systems showed that all of them were efficient on this purpose. The highest oxygen consumption rates during these reactions were observed in the Fe2+ initiated reactions. The LCC inhibited the peroxidation reactions when added to the reaction medium at concentrations higher than 0.3 mg/mL. However, prolonging the reactions up to 72h with LCC at 1 mg/mL showed that it was depolymerized. The lignin degradation routes involved depolymerization or simple side chain and free-phenolic structure oxidations. When the reactive system was based on the use of MnP/Mn+2/H2O2/linoleic acid, some lignin degradation routes were demonstrated and they included Cα-oxidation, as well as β-O-4 and/or Cα-Cβ cleavages. These results corroborate previous findings published for the action of C. subvermispora in vivo. One exception was the simple Cα oxidation that was observed for the in vitro reactions, but was ruled out by previous works that were based on the characterization of residual lignins extracted from wood samples biotreated by C. subvermispora (in vivo system). The current results permitted to conclude that several mimetic systems were able to initiate linoleic acid peroxidation in vitro. When these reactions were performed in the presence of lignin (LCC or milled E. grandis) it was possible to show the occurrence of several lignin transformation reactions that could be exploited, for example, in pulp bleaching processes.

Summary Decay of birch (Betul papyrifera), aspen (Poplus tremuloides) and loblolly pine (Pinus taeda) wood by 19 isolates of Phanerochaete chrysoporium showed considerable variation in percent weight loss and loss of lignin and wood sugars. Birch and aspen woods are degraded to a greateer extent than loblolly pine wood. Among the isolates tested, a great deal of variation was observed in their ability to preferentiall y degrade lignin. Isolate BKM-F-1767 resulted in the greatest loss of lignin on the deciduous woods tested. Many isolates of P. chrysoporium removed all cell wall components causing a nonselective type fo white rot. In contreast, nine isolates of Ceriporiopsis subvermispora caused moderate weight losses and perferential degradation of lignin in aspen, birch and loblolly pine wood. Less variation among isolates was observed in the cell wall components removed from all woods tested. Lignin losses ranged from 19 to 38% in loblolly pine and 50-80% in aspen and birch wood. Polyacrylamide gel electrophoresis (SDS-PAGE) showed a wide variety of band patterns for extracellular enzymes among isolates of P. chrysosporium, but relatively uniform protien bands among isolates of C. subvermispora . Only a few isolates of P. chrysosporium appear to be strains that preferentially removed large amounts of lignin from wood, whereas all isolates of C. subvermisproa tested are selective lignin degraders on deciduous as well as coniferous wood.

Wood is the main raw material used in the pulp and paper industry. It is a material that presents heterogeneous structure and complex composition, which results in a relatively resistant material to the biodegradation process. In the present review, we attempted to summarize the structural characteristics of wood and describe the chemical nature of its major components to, afterwards, comment about its biodegradation. The role of the enzyme manganese peroxidase in the lignin degradation by a selective white-rot fungus, Ceriporiopsis subvermispora, was highlighted.

Os fungos de decomposição branca são organismos capazes de degradar os materiais lignocelulósicos de maneira eficiente através de reações catalisadas por enzimas hidrolíticas e oxidativas ou ainda por meio de metabólitos de baixa massa molar que iniciam o processo degradativo. Ceriporiopsis subvermispora é um fungo de decomposição branca que se destaca como um dos mais seletivos na degradação da lignina, possuindo grande potencial de aplicação tecnológica. Este fungo é capaz de iniciar reações de peroxidação de lipídeos a partir da formação de íons Mn3+ oriundos da ação da enzima manganês peroxidase (MnP) sobre Mn2+. No presente estudo avaliou-se a secreção de enzimas oxidativas por C. subvermispora agindo sobre madeira de Eucalyptus grandis em cultivos suplementados ou não com milhocina e milhocina e glicose. Os extratos enzimáticos obtidos a partir desses cultivos apresentaram elevada atividade de MnP, principalmente quando o meio foi suplementado com milhocina. Os mesmos extratos foram capazes de peroxidar ácido linoleico in vitro. Nessas reações ocorreu um acúmulo de substâncias reativas com ácido tiobarbitúrico (TBARS) que foi proporcional ao nível de MnP existente nos extratos. A taxa de consumo de oxigênio nessas reações também foi proporcional aos teores de MnP presente nos extratos. A caracterização da madeira biotratada indicou que as maiores perdas de massa e de lignina ocorreram nos cultivos suplementados, sendo que aparentemente há uma correlação entre a extensão da degradação de madeira e lignina com a atividade peroxidativa dos extratos. Lacases foram detectadas somente em baixos níveis nos extratos enzimáticos oriundos da madeira. Por outro lado, cultivos em meio líquido composto de 2% de extrato de malte e 0,2% de extrato de levedura induziram a produção de lacases e nenhuma secreção de MnP. Também as lacases foram capazes de peroxidar ácido linoleico, desde que na presença de um mediador, o ácido _-hidróxi-benzóico. De qualquer forma, durante a biodegradação de E. grandis por C. subvermispora, lacase foi detectada somente no início dos cultivos e mesmo assim em níveis bastante baixos o que significa que a participação de lacase na peroxidação de lipídeos durante a biodegradação de E. grandis por C. subvermispora deve ser pouco significante. The white-rot fungi are able to degrade lignocellulosic materials in an efficient way through reactions catalyzed by hydrolytic and oxidative enzymes or still by means of low molar mass metabolites that begin the degradative process. Ceriporiopsis subvermispora is a white-rot fungus that stands out as one of the most selective in lignin degradation, possessing great potential for technological applications. This fungus is able to initiate lipid peroxidation reactions based on the Mn3+ produced by the catalytic action of the enzyme manganese peroxidase (MnP) on Mn2+ ions. In the present study, the secretion of oxidative enzymes by C. subvermispora was evaluated in cultures containing Eucalyptus grandis wood chips and some supplements such as corn steep liquor and corn steep liquor plus glucose. The enzymatic extracts obtained from these cultures contained high MnP activity, mainly when corn steep liquor was used as a culture supplement. The same extracts were able to peroxidize linoleic acid in vitro. In these reactions, thiobarbituric acid reactive substances (TBARS) accumulated over reaction time and the TBARS levels were proportional to the MnP present in the extracts. The rate of oxygen consumption in these reactions was also proportional to the level of MnP present in the extracts. The characterization of the biotreated wood indicated that the largest lignin and wood mass losses were observed in supplemented cultures, with an apparent correlation between the peroxidative activity in the extracts and the extension of the wood and lignin degradation. Laccase was detected only in low levels in the enzymatic extracts originating from the wood-containing cultures. On the other hand, submerged cultures in which the liquid medium was composed of 2% malt-extract and 0,2% yeast extract induced laccase production without simultaneous secretion of MnP. The laccase active extract was also able to peroxidize linoleic acid, since in the presence of a mediator (_-hydroxy-benzoic acid). Anyway, during E. grandis biodegradation by C. subvermispora, laccase was only detected in the beginning of the cultivations and even so at low levels suggesting that laccase were not involved in lipid peroxidation during the E. grandis wood biodegradation by C. subvermispora.

Ceriporiopsis subvermispora was used for biochemical pulping of agricultural residues and the results were compared with chemical pulping. Independent variables were screened by Plackett-Burman and optimized by full factorial experimental designs. Biological treatment of rice, wheat and barley straw samples resulted in decrease of the kappa number of these straws by 34%, 21% and 19%, respectively, as compared with controlled samples. The tensile strength and burst factor of hand sheets produced from rice straw were increased by 51% and 33% as compared with the control straws. The tensile strength and burst factor of hand sheets produced from wheat straws were improved by 67% and 36%, these variables for barely straws were 36.7% and 45%, respectively. Although the delignification of wheat and barley straws are not as efficient as chemical process, but the quality of papers produced by biochemical pulping of straws were excellent.

Ceriporiopsis subvermispora is a white-rot basidiomycete that degrades lignin selectively and often is used in biopulping. Development of appropriate experimental procedures for recovery and determination of enzymes produced by C. subvermispora during biopulping is necessary to evaluate the role of each enzyme in this process. In this work, enzymes produced during biopulping were extracted and determined by using a series of substrates. Manganese-dependent peroxidases (MnP) predominated over laccases, which were of minor significance. The lack of laccases was further confirmed by assaying several substrates such as syrindaldazine, o-dianisidine and ABTS. A basal oxidation of syringaldazine and phenol red in the absence of exogenous Mn2+ led to the assumption that the extracts contained some Mn-independent peroxidase or enough Mn2+ to initiate MnP catalyzed reactions. The presence of Mn2+ was evidenced since phenol red was not oxidized by a 16-h-dialysed extract. Addition of 111 μM of Mn2+ to the reaction medium restored the oxidative activity. Lignin peroxidase activity was not detected in the extracts based on the Azure B assay. Cellulase activities were very low while xylanases predominated. The xylanolytic complex was deficient in beta-xylosidase. Successive extractions were performed for a quantitative recovery of the enzymes adsorbed on wood chips. The first 4-h extraction recovered an average of 50% of the total enzymatic activities.

Indexación: Scopus. Genetic analyses of the lignin-degrading fungus Ceriporiopsis subvermispora is complicated by a dikaryotic nuclear condition and the absence of spore forms. Previous investigations had identified a family of closely related sequences encoding manganese peroxidase (MnP), but the relationship between genes and allelic variants could not be experimentally established. Addressing this issue, homokaryotic derivatives of C. subvermipora strain FP105752 were isolated from regenerated protoplasts. Designated CsA and CsB, their homokaryotic nature was established by polymerase chain reaction amplification and sequence analysis of the allelic variants of three MnP genes. Isoelectrofocusing revealed fewer MnP isoenzymes in filtrates of homokaryon cultures relative to the parental strain. The homokaryotic strains will simplify genetic analyses, particularly the identification of new genes. https://academic-oup-com.recursosbiblioteca.unab.cl/femsle/article/199/1/91/553870

A new method to facilitate rapid screening of lignin-degrading microorganisms was developed. Fungal strains are cultivated in tissue culture plates containing 14 C-ring-labeled dehydrogenation polymerizate (DHP) (synthetic lignin). Evolved 14 CO 2 is trapped in barium-saturated filter paper and is detected by exposing the paper to X-ray film. Analysis of the autoradiograms, carried out by density measurement with an image analysis program, allows for a semiquantitative estimation of the amount of 14 CO 2 evolved. The method is especially useful for screening for new, powerful lignin-degrading strains in both man-made and natural environments. It eliminates the need for special equipment for their cultivation and trapping of 14 CO 2 as well as laborious sample analysis. The method has in this study been used to test three novel fungal isolates and a laccaseless mutant of the basidiomycete Pycnoporus cinnabarinus . Their ligninolytic capacities were compared with those of the potent lignin degrader Ceriporiopsis subvermispora.