• Journal of Microbiology
• /
• v.43 no.6
• /
• pp.487-492
• /
• 2005

This study demonstrated that the brown rot basidiomycete Fomitopsis palustris was able to degrade crystalline cellulose (Avicel). This fungus could also produce the three major cellulases (exoglucanases, endoglucanases, and $\beta-glucosidase$) when the cells were grown on $2.0\%$ Avicel. Avicel degraded by F. palustris showed a decrease in relative crystallinity from $83\%\;to\;78.5\%$ after 14 days of incubation. The characterization study indicated that optimum pH was 4.5 and optimum temperature was $70^{\circ}C$ for exoglucanase (cellobiohydrolase) activity. Hydrolysis of Avicel by the crude enzyme from F. palustris yielded 1.6 mg/ml of glucose after 43 h, which corresponded to a cellulose conversion degree of $3.2\%$. Therefore, this study revealed for the first time that the brown rot basidiomycete F. palustris produces cellulases capable of yielding soluble sugars from crystalline cellulose.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.4
• /
• pp.347-358
• /
• 2011

Fomitopsis palustris, a brown-rot basidiomycete, causes the most destructive type of decay in wooden structures. In spite of its great economic importance, very little information is available at the molecular level regarding its complex decay process. To address this, we generated over 3,000 expressed sequence tags (ESTs) from a cDNA library constructed from F. palustris. Clustering of 3,095 high-quality ESTs resulted in a set of 1,403 putative unigenes comprising 485 contigs and 918 singlets. Homology searches based on BlastX analysis revealed that 78% of the F. palustris unigenes had a significant match to proteins deposited in the nonredundant databases. A subset of F. palustris unigenes showed similarity to the carbohydrateactive enzymes (CAZymes), including a range of glycosyl hydrolase (GH) family proteins. Some of these CAZyme-encoded genes were previously undescribed for F. palustris but predicted to have potential roles in biodegradation of wood. Among them, we identified and characterized a gene (FpCel45A) encoding the GH family 45 endoglucanase. Moreover, we also provided functional classification of 473 (34%) of F. palustris unigenes using the Gene Ontology hierarchy. The annotated EST data sets and related analysis may be useful in providing an initial insight into the genetic background of F. palustris.

• Journal of Forest and Environmental Science
• /
• v.24 no.1
• /
• pp.53-59
• /
• 2008

Extracellular enzyme activities of Fomitopsis palustris were determined by the particle sizes, culture periods and concentrations of wood particle substrate which was mixture of 4 domestic coniferous woods, such as Pinus densiflora, Larix leptolepsis, Pinus koraiensis, and Pinus rigida. The results showed that the culture conditions had an effect on the secretion of most of the extracellular enzymes from Fomitopsis palustris in the mixed wood particle substrate. :The optimal culture conditions for enzyme activities were 80~100 mesh in wood particle size, 7.5% in concentrations of wood substrate, and 4~8 weeks in culture period.

• Journal of the Korean Wood Science and Technology
• /
• v.35 no.6
• /
• pp.159-165
• /
• 2007

An extracellular xylanase from the brown-rot fungus Fomitopsis palustris grown on rice straw culture was purified to a single protein band. On SDS-PAGE, the molecular mass of purified enzyme was estimated to be about 43 kDa. The amino acid sequence of the proteolytic fragments showed high homology with fungal glycoside hydrolase family 10 xylanases. The $K_m$, $K_{cat}$ and $V_{max}$ for birch xylan were $31mg/m{\ell}$, $2.3{\times}10^4/min$ and 252.3 U/mg, respectively. The optimal activity of the purified xylanase from F palustris was observed at pH 4.0~5.0 and $70^{\circ}C$.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.3
• /
• pp.404-409
• /
• 2008

The brown-rot basidiomycete Fomitopsis palustris is known to degrade crystalline cellulose (Avicel) and produce three major cellulases, exoglucanases, endoglucanases, and ${\beta}$-glucosidases. A gene encoding endoglucanase, designated as cel12, was cloned from total RNA prepared from F. palustris grown at the expense of Avicel. The gene encoding Cel12 has an open reading frame of 732 bp, encoding a putative protein of 244 amino acid residues with a putative signal peptide residing at the first 18 amino acid residues of the N-terminus of the protein. Sequence analysis of Cel12 identified three consensus regions, which are highly conserved among fungal cellulases belonging to GH family 12. However, a cellulose-binding domain was not found in Cel12, like other GH family 12 fungal cellulases. Northern blot analysis showed a dramatic increase of cel12 mRNA levels in F. palustris cells cultivated on Avicel from the early to late stages of growth and the maintenance of a high level of expression in the late stage, suggesting that Cel12 takes a significant part in endoglucanase activity throughout the growth of F. palustris. Adventitious expression of cel12 in the yeast Pichia pastoris successfully produced the recombinant protein that exhibited endoglucanase activity with carboxymethyl cellulose, but not with crystalline cellulose, suggesting that the enzyme is not a processive endoglucanase unlike two other endoglucanases previously identified in F. palustris.

• Journal of the Korean Wood Science and Technology
• /
• v.35 no.6
• /
• pp.91-99
• /
• 2007

Activities of the extracellular enzyme from Fomitopsis palustris, a brown-rot fungi, and by which crystallinity changes of cellulose in the various softwoods, such as Larix leptolepsis, Finns rigida, Finns koraiensis and Finns densiflora by liquid culture, were investigated. Activity of Cellobiohydrolase (CBH) from F. palustris was detected in the every test softwoods culture, showing activities of the Endoglucanase (EG), $\beta$-glucosidase (BGL) and $\beta$-1,4-xylosidase (BXL). It was shown high enzyme activities in the sapwood culture than heartwood of the same wood species, However, the enzyme activities in most of test wood cultures increased with longer incubation time, indicating a possibility of intermix sapwood and heartwood for degradation process by enzyme. Also it was shown that protein patterns of the extracellular enzyme from F. palustris in wood particle substrate of the several domestic softwoods were similar with each other wood species, which suggested the possibility of mixing all softwoods in saccharification by enzyme from F. palustris. Crystallinity reduction value of cellulose by F. palustris was 4.2~20.4% in 4 weeks cultivation, 12.9~28.9% in 8 weeks.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.5
• /
• pp.800-805
• /
• 2007

Two endoglucanases with processive cellulase activities, produced from Fomitopsis palustris grown on 2% microcrystalline cellulose(Avicel), were purified to homogeneity by anion-exchange and gel filtration column chromatography systems. SDS-PAGE analysis indicated that the molecular masses of the purified enzymes were 47 kDa and 35 kDa, respectively. The amino acid sequence analysis of the 47-kDa protein(EG47) showed a sequence similarity with fungal glycoside hydrolase family 5 endoglucanase from the white-rot fungus Phanerochaete chrysosporium. N-terminal and internal amino acid sequences of the 35-kDa protein(EG35), however, had no homology with any other glycosylhydrolases, although the enzyme had high specific activity against carboxymethyl cellulose, which is a typical substrate for endoglucanases. The initial rate of Avicel hydrolysis by EG35 was relatively fast for 48 h, and the amount of soluble reducing sugar released after 96 h was $100{\mu}g/ml$. Although EG47 also hydrolyzed Avicel, the hydrolysis rate was lower than that of EG35. Thin layer chromatography analysis of the hydrolysis products released from Avicel indicated that the main product was cellobiose, suggesting that the brown-rot fungus possesses processive EGs capable of degrading crystalline cellulose.

• Journal of the Korean Wood Science and Technology
• /
• v.38 no.3
• /
• pp.262-273
• /
• 2010

In the enzymatic hydrolysis of rice straw and wood meals using extra-cellular enzymes from Fomitopsis palustris, key factors which enhanced the sugar conversion yield were investigated in this work, such as enzyme production and enzyme reaction conditions, surfactant effects, and the surface structure of substrates. F. palustris cultured with softwood mixture produced 12.0 U/$m{\ell}$ for endo-${\beta}$-1,4-gulcanase (EG), 116.68 U/$m{\ell}$ for ${\beta}$-glucosidase (BGL), 18.82 U/$m{\ell}$ for cellobiohydrolase (CBH), and 13.33 U/$m{\ell}$ for ${\beta}$-xylosidase (BXL). These levels of BGL, CBH, and BXL activities were two to four folds more than enzyme activities of F. palustris cultured with rice straw. The optimum reaction conditions of cellulase-RS which produced by F. palustris with rice straw and cellulase-SW which produced by F. palustris with softwood mixture were pH 5.0 at $45^{\circ}C$ and pH 5.0 at $50^{\circ}C$, respectively. The sugar conversion yield of cellulase-SW had the highest value of $40.6{\pm}0.6%$ within 72 h when rice straw was used as substrate. By adding 0.1% Tween 20 (w/w-substrate), the sugar conversion yield of rice straw was increased to 44%, which was about four fifths sugar conversion yield of commercial enzyme, Celluclast 1.5L (Novozyme A/S). A low crystallinity and an intensive fibril surface observed by the scanning electron microscope may explain the high sugar conversion yield of rice straw.

• 한국신재생에너지학회:학술대회논문집
• /
• 2006.11a
• /
• pp.529-532
• /
• 2006

Recently the production of ethanol from lignocecllulosics has received much attention due to immense potential for conversion of renewable biometerials into biofuels and chemicals. Fomitopsis palustris causes a typycal brown-rot and is unusual in that it rapidly depolymerize the cellulose in wood without removing the surrounding lignin that normally prevents microbial attack. This study demonstrated that the brown rot basidiomycete F. palustris was able to degrade crystalline cellulose. This fungus could also produce the three major cellulases (BGL, EXG and EG) when the cells were grown on 2.0% Avicel. The fungus was able to degrade both the crystalline and amorphous forms of cellulose from woody biomasses. Moreover, we found that this fungus has the processive EG like CBH which are able to degrade the crystalline region of cellulose. To establish the cellulase system in relation with degradation of woody biomass, we performed that purification, characterization and molecular cloning of a BGL, EGs and GLA from F. palustris grown on Avicel.

• Journal of the Korean Wood Science and Technology
• /
• v.44 no.6
• /
• pp.880-889
• /
• 2016

To evaluate the resistance of wood to decay caused by fungi, sapwood and heartwood of red pine (Pinus densiflora) and sapwood and heartwood of larch (Larix kaempferi) were conducted. Wood samples were immersed for 96 h in pyroligneous liquor. Then, the brown-rot fungus, Fomitopsis palustris, was used to examine the decay resistance of red pine and larch. Weight and density of wood from the all conditions increased after immersion treatment. Weight loss after decay resistance test was also dropped with a immersion treatment. The lowest weight loss indicated at immersion-treated heartwood of larch. Immersion treatment using pyroligneous liquor effectively increased the resistance of wood to decay caused by fungi.