Phlebia Brevispora Research Articles

Influence of metals on lignocellulolytic enzyme production and metal removal by Phlebia brevispora in a wheat straw based fixed‐bed bioreactor

AbstractHeavy metals contamination poses a significant environmental threat, which requires the development of eco‐friendly bioremediation techniques. Present research work was conducted using wheat straw in submerged conditions for the growth of white rot fungus, Phlebia brevispora in the presence of varying concentrations of Cd, Pb, Ni, and Cr. The lignocellulolytic enzyme production ability of fungus was monitored. A wheat straw‐based fixed‐bed bioreactor was designed to treat metal‐containing water in continuous mode. The fungus showed a positive influence on laccase production, cellulase production, and lignin peroxidase activity. As observed through atomic absorption spectroscopy (AAS), Cr removal efficiency was 78%–80%, regardless of the initial metal concentration. The Cr was also found on the surface of fungal mycelium as per the results obtained from SEM–EDX. The continuous bioreactor achieved 98%–99% metal removal, making it a natural and cost‐effective solution for metal removal from wastewater using P. brevispora.

Conversion of lignocellulosic biomass: Production of bioethanol and bioelectricity using wheat straw hydrolysate in electrochemical bioreactor

The present study evaluated efficiency of wheat straw (WS) hydrolysate obtained through fungal pre-treatment to produce ethanol and electricity in an electrochemical bioreactor. Three white rot fungi Phanerochaete chrysosporium, Phlebia floridensis and Phlebia brevispora were used to degrade WS for hydrolysate preparation, Lignocellulolytic enzyme production was also monitored during the pretreatment. Yeast Pichia fermentans was allowed to ferment all three hydrolysates up to 12 days. The yeast showed maximum electrochemical response as open circuit voltage (0.672 V), current density 542.42 mA m−2, and power density of 65.09 mW m−2 on 12th day in the hydrolysate prepared using Phlebia floridensis. Maximum ethanol production of 9.2% (w/v) was achieved on 7th day with a fermentation efficiency of about 62.1%. Further, the coulombic efficiency improved from 0.06 to 1.46% during 12 days of the experiment. Thus, the results indicated towards the possible conversion of lignocellulosic biomass into bioethanol along with bioelectricity generation.

Electroactive biofilm development on carbon fiber anode by Pichia fermentans in a wheat straw hydrolysate based microbial fuel cell

Abundantly available lignocellulosic biomass may be used as feed stock for various applications including microbial fuel cells. However, a pre-treatment of such biomass is always required for an efficient operation. In the present study, a double-chambered fuel cell was developed to evaluate the biofilm formation along with electricity production ability of Pichia fermentans using wheat straw hydrolysate. The wheat straw hydrolysate was prepared by degrading the wheat straw using white rot fungi, Phlebia brevispora, Phanerochaete chrysosporium, and Phlebia floridensis. This hydrolysate primarily consisted of sugars, protein, phenols, and enzymes and used as anolyte in fuel cell. The hydrolysate as anolyte facilitated the growth of yeast Pichia fermentans and development of biofilm. Maximum electrochemical response of 20.13 ± 0.052 mW m−2 and 20.42 ± 0.071 mW m−2 was recorded in hydrolysate prepared by using P. floridensis and P. chrysosporium, respectively. Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM) of the biofilm revealed the typical characteristics along with significant exopolysaccharide (EPS) production. The study demonstrated active biofilm formation by yeast using wheat straw hydrolysate, which further indicates the application of such organisms in the development of lignocellulosic biomass based systems including the electrochemical one.

Development of wheat straw based catholyte for power generation in microbial fuel cell

Microbial fuel cell has been extensively used for electricity generation. A dual chambered microbial fuel cell was designed to degrade wheat straw by white rot fungi, Phlebia floridensis and Phlebia brevispora in cathode chamber. Pichia fermentans was used as exoelectrogenic yeast in anodic chamber. Lignocellulolytic enzymes including cellulase and laccase were efficiently produced by both the fungi in cathode chamber during 30 days of experiment. Concentrations of reducing sugars and phenolics in the catholyte increased during wheat straw degradation. Maximum laccase (9.035 CU ml−1) was produced by P. floridensis and a maximum open circuit voltage (0.504 V) was achieved on 9th day. P. brevispora produced maximum laccase upto 4.77 CU ml−1 and maximum open circuit voltage (0.496 V) was recorded on 17th day. Power density followed the pattern of open circuit voltage and a maximum power density of 33.19 mW m−2 and 12.90 mW m−2 was achieved by P. floridensis and P.brevispora, respectively. The degradation of wheat straw improved the efficiency of catholyte by improving electricity generation in a yeast fuel cell. Thus, the setup explores the potential of agricultural waste to be used in such sustainable and eco-friendly technology to produce bio-electricity.

Potency of Phlebia species of white rot fungi for the aerobic degradation, transformation and mineralization of lindane.

The widespread use of the organochlorine insecticide lindane in the world has caused serious environmental problems. The main purpose of this paper is to investigate the potency of several Phlebia species of white rot fungi to degrade, transform and mineralize lindane, and to provide the feasibility of using white rot fungi for bioremediation at contaminated sites. Based on tolerance experiment results, Phlebia brevispora and Phlebia lindtneri had the highest tolerance to lindane and were screened by degradation tests. After 25 days of incubation, P. brevispora and P. lindtneri degraded 87.2 and 73.3% of lindane in low nitrogen medium and 75.8 and 64.9% of lindane in high nitrogen medium, respectively. Several unreported hydroxylation metabolites, including monohydroxylated, dehydroxylated, and trihydroxylated products, were detected and identified by GC/MS as metabolites of lindane. More than 10% of [14C] lindane was mineralized to 14CO2 by two fungi after 60 days of incubation, and the mineralization was slightly promoted by the addition of glucose. Additionally, the degradation of lindane and the formation of metabolites were efficiently inhibited by piperonyl butoxide, demonstrating that cytochrome P450 enzymes are involved in the fungal transformation of lindane. The present study showed that P. brevispora and P. lindtneri were efficient degraders of lindane; hence, they can be applied in the bioremediation process of lindane-contaminated sites.

Growth management of white-rot fungus Phlebia brevispora improved degradation of high-molecular-weight polycyclic aromatic hydrocarbons.

The effect of co-culturing white-rot fungus Phlebia brevispora with growth-promoting bacterial strains Enterobacter sp. TN3W-14 and Pseudomonas sp. TN3W-8 on the degradation of polycyclic aromatic hydrocarbons (PAHs) was evaluated in liquid culture. In axenic cultures, P. brevispora strains TN3F and TMIC33929 showed high degradation of phenanthrene (> 98%) within 15days, and degraded 65% and 63% of pyrene, and 12% and 8% of benzo(a)pyrene, respectively. This low level of degradation ability toward benzo(a)pyrene was improved significantly by co-culturing the fungi with a mixture of bacterial strains TN3W-8 and TN3W-14 (mixed bacterial co-culture; MBC). Within 15days, P. brevispora TN3F with MBC achieved about 86% pyrene and 53% benzo(a)pyrene degradation; P. brevispora TMIC33929 with MBC showed 92% pyrene and 72% benzo(a)pyrene degradation. The MBC alone degraded little PAH, as its growth was inhibited by PAH; however, its co-culture with P. brevispora improved mycelial growth of the fungus, which led to improved degradation of the PAHs. A possible dihydrodiol metabolite of pyrene from fungal cultures suggests that hydroxylation was the first step in the degradation of pyrene by P. brevispora.

Characteristics of White-rot Fungus Phlebia brevispora TMIC33929 and Its Growth-Promoting Bacterium Enterobacter sp. TN3W-14 in the Decolorization of Dye-Contaminated Water.

The dye decolorization potential of the white-rot fungus Phlebia brevispora TMIC33929 when grown alone or in coculture with its growth-promoting bacterium Enterobacter sp. TN3W-14 was evaluated in low nitrogen liquid medium at different pHs. Axenic fungus removed a similar amount of Congo red and crystal violet at pH 4.5 and 7.0, respectively. The bacterium alone achieved only slightly better decolorization of crystal violet than the fungus at pH 9.0. Compared with axenic fungus, cocultures provided no increased crystal violet removal but achieved higher removal of crystal violet in mixed dye at all pHs, and the best-mixed dye decolorization at pH 9.0. Unlike bacterial growth on dyes, growth of fungal mycelia was not inhibited by the dyes at all pH but the cocultures gave comparably higher mycelial growth.

Indirect Bacterial Effect Enhanced Less Recovery of Neonicotinoids by Improved Activities of White-Rot Fungus Phlebia brevispora.

Bacterial strains that improve mycelial morphology and growth of white-rot fungi in liquid medium could enhance the impact of white-rot fungi towards lesser recovery of neonicotinoids when cocultured. This was demonstrated by the recovery of clothianidin and acetamiprid from cocultures of the white-rot fungus Phlebia brevispora strains with two mycelial-growth-promoting bacteria, Enterobacter sp. TN3W-14 and Pseudomonas sp. TN3W-8. Clothianidin recovery from cocultures of white-rot fungi and bacteria was over 40% lower than that from axenic microbial cultures and mixed-bacterial cultures. About 20% less acetamiprid was equally recovered from both TMIC33929+TN3W-14 cocultures and mixedbacterial cultures than from axenic fungal and bacterial cultures.

Biodegradation and bioconversion of endrin by white rot fungi, Phlebia acanthocystis and Phlebia brevispora

Endrin is persistent organic pollutants that contaminate soil in many parts of the world. In this study, endrin was used as the substrate for a degradation experiment with the white rot fungi of the genus Phlebia . The results of tolerance test showed that the tolerance level of Phlebia acanthocystis and Phlebia brevispora to endrin was higher than that of other fungi, and the tolerance coefficient of both strains to 1.0 mg/L endrin exceeded 0.9 in solid PDA medium. P. acanthocystis and P. brevispora could degrade endrin efficiently in pure culture, especially P. acanthocystis had the highest degradability of more than 80% after 20 d incubation. Compared with low-nitrogen medium, PDB medium is more suitable for the biodegradability of two fungi. Several hydroxylated products such as 8-hydroxyendrin and two monohydroxyendrin were detected, indicating that endrin was initially branched to different monohydroxylated products in fungal degradation. Moreover, a carboxylic acid product was obtained from P. acanthocystis culture, suggesting that the carboxylation reaction occurred in bioconversion of endrin. The fungal cytochrome P450 enzymes play significant role in the in the initial hydroxylation process on endrin degradation. This is the first report that endrin is converted to hydroxylated and carboxylated metabolites by microorganisms. • Phlebia acanthocystis and P. brevispora can tolerate endrin of different concentrations. • Both strains showed excellent ability to degrade endrin in PDB and HCLN media. • Endrin was converted into several hydroxylated and carboxylated products. • Fungal Cyt P450 is involved in the initial hydroxylation of endrin.

Antioxidant Potential and Extracellular Auxin Production by White Rot Fungi.

Selective lignin degrading white rot fungi viz. Phanerochaete chrysosporium, Phlebia brevispora, and Phlebia floridensis were selected to evaluate antioxidant potential and auxin (indole acetic acid) production in complex and synthetic medium. Antioxidant potential of these fungi was tested against different free radicals including 2, 2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide, ferrous ion, and ferric ion along with total phenolic content. All the fungal strains produce phenolics ranging from 5.2 to 16.7mg/ml and demonstrated various free radical and metal ion scavenging activity. Growth medium significantly affected all the activities. Almost similar antioxidant activity (~ 72% DPPH scavenging activity) was demonstrated by all the fungi in yeast extract glucose medium; however, the activity was lower in Czapek dox's medium (from 60 to 45%). Indole acetic acid production was maximum in P. brevispora (31μg/ml), which was closely followed by P. chrysosporium and P. floridensis. The extracts did not show any mutagenic or cytotoxic effect. Thus, these white rot fungi highlight their significance as a new source for the prompt production of extracellular antioxidants and auxin.

Bacterial strains isolated from cedar wood improve the mycelial growth and morphology of white rot fungus Phlebia brevispora on agar and liquid medium

Growth enhancement of white rot fungus Phlebia brevispora TN3F by two bacterial strains TN3W-8 and TN3W-14 which showed high identity with genus Pseudomonas and Enterobacter, respectively, was investigated in liquid and solid media at different glucose concentrations. The white rot fungus P. brevispora TN3F and the bacterial strains were isolated from same white rot-decayed cedar wood from the Tano Forest Science Station at University of Miyazaki. Confrontational assay between the fungus and the bacterial strains revealed a fungal growth enhancement phenomenon, in the direction of the bacterial cells, even without direct contact. This enhancement increased with decrease of glucose concentration, 0.5% glucose offering comparatively the most mycelial growth. In potato extract (PE) medium at 0.5% glucose concentration, along with mycelial growth enhancement and morphological change of P. brevispora TN3F, growth and morphological enhancement of fungus TMIC33929, another strain of P. brevispora, was achieved by the bacterial strain TN3W-14, although TMIC33929 originated from a different environment. Outside few reports of a positive interaction between white rot fungi and other microorganisms, this is a first report of such positive interaction in liquid medium. Furthermore, this also is the first report of a single bacterium having potential to positively affect mycelia growth of different strains of a white rot fungus.

Biological pretreatment of corn stover with Phlebia brevispora NRRL-13108 for enhanced enzymatic hydrolysis and efficient ethanol production.

Biological pretreatment of lignocellulosic biomass by white-rot fungus can represent a low-cost and eco-friendly alternative to harsh physical, chemical, or physico-chemical pretreatment methods to facilitate enzymatic hydrolysis. In this work, solid-state cultivation of corn stover with Phlebia brevispora NRRL-13018 was optimized with respect to duration, moisture content and inoculum size. Changes in composition of pretreated corn stover and its susceptibility to enzymatic hydrolysis were analyzed. About 84% moisture and 42 days incubation at 28°C were found to be optimal for pretreatment with respect to enzymatic saccharification. Inoculum size had little effect compared to moisture level. Ergosterol data shows continued growth of the fungus studied up to 57 days. No furfural and hydroxymethyl furfural were produced. The total sugar yield was 442 ± 5 mg/g of pretreated corn stover. About 36 ± 0.6 g ethanol was produced from 150 g pretreated stover per L by fed-batch simultaneous saccharification and fermentation (SSF) using mixed sugar utilizing ethanologenic recombinant Eschericia coli FBR5 strain. The ethanol yields were 32.0 ± 0.2 and 38.0 ± 0.2 g from 200 g pretreated corn stover per L by fed-batch SSF using Saccharomyces cerevisiae D5A and xylose utilizing recombinant S. cerevisiae YRH400 strain, respectively. This research demonstrates that P. brevispora NRRL-13018 has potential to be used for biological pretreatment of lignocellulosic biomass. This is the first report on the production of ethanol from P. brevispora pretreated corn stover. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:365-374, 2017.

Simultaneous Degradation of Polycyclic Aromatic Hydrocarbons by Attractive Ligninolytic Enzymes from Phlebia brevispora KUC9045

The hazards associated with the polycyclic aromatic hydrocarbons (PAHs) are known to be recalcitrant by their structure, but white rot fungi are capable of degrading recalcitrant organic compounds. Phlebia brevispora KUC9045 isolated from Korea was investigated its effi-ciency of degradation of four PAHs, such as phenanthrene, anthracne, fluoranthene, and pyrene. And the species secreted extracellular laccase and MnP(Manganese dependent peroxidase) during degradation. P. brevispora KUC9045 demonstrated effective degradation rates of phenanthrene (66.3%), anthracene (67.4%), fluoranthene (61.6%), and pyrene (63.3%), respectively. For enhancement of degradation rates of PAHs by the species, Remazol Brilliant Blue R (RBBR) was preferentially supplemented to induce ligninolytic enzymes. The biodegradation rates of the three PAHs including phenanthrene, fluoranthene, and pyrene were improved as higher concentration of Remazol Brilliant Blue R was supplemented. However, anthracene was degraded with the highest rate among four PAHs after two weeks of the incubation without RBBR addition. According to the previous study, RBBR can be clearly decolorized by P. brevispora KUC9045. Hence, the present study demonstrates simultaneous degradation of dye and PAHs by the white rot fungus. And it is considered that the ligninolytic enzymes are closely related with the degradation. In addition, it indicated that dye waste water might be used to induce ligninolytic enzymes for effective degradation of PAHs.

Decolorizing an anthraquinone dye by Phlebia brevispora: Intra-species characterization.

Remazol brilliant blue R (RBBR) is an anthraquinone dye derived from anthracene that is decolorized by a white rot fungus, Phlebia brevispora. Interestingly, P. brevispora produces two phenomena of yellowish and pinkish colors during the degradation of RBBR. Here, we characterized the decolorization of RBBR by P. brevispora. The fungus was significantly different between the two colors via UV spectrophotometry, and the morphology of the hyphae observed in the respective color culture was also entirely different. Moreover, both of the two ligninolytic enzymes, laccase and manganese-dependent peroxidase (MnP), were remarkably stimulated in the yellowish culture at the beginning of the decolorization. It is possible that the RBBR decolorizing mechanism might be primarily related to the amount of laccase and MnP produced in the yellowish culture. Thus, the decolorized color may be rapidly estimated at initial period of incubation. In addition, GeneFishing technology revealed that two genes were differentially expressed in yellowish culture.

Biological pretreatment of corn stover with white-rot fungus for improved enzymatic hydrolysis

Biological pretreatment of lignocellulosic biomass by white-rot fungus can represent a low-cost and eco-friendly alternative to harsh physical, chemical or physico-chemical pretreatment methods to facilitate enzymatic hydrolysis. However, fungal pretreatment can cause carbohydrate loss and it is, therefore, necessary to use the appropriate fungal strain-biomass type combination. In this work, 26 white-rot fungal strains were evaluated under solid state cultivation at 74% moisture level at 28 °C for 30 days for fermentable sugar production from corn stover after enzymatic hydrolysis of each fungal pretreated corn stover using a cocktail of 3 commercial enzyme (cellulase, β-glucosidase, hemicellulase) preparations. The best result was obtained with Cyathus stercoreus NRRL-6573 which gave a sugar yield of 394 ± 13 mg/g of pretreated stover followed by Pycnoporus sanguineus FP-10356-Sp (393 ± 17 mg/g) and Phlebia brevispora NRRL-13108 (383 ± 13 mg/g).

A survey of genes encoding H2O2-producing GMC oxidoreductases in 10 Polyporales genomes

The genomes of three representative Polyporales (Bjerkandera adusta, Phlebia brevispora and a member of the Ganoderma lucidum complex) recently were sequenced to expand our knowledge on the diversity and distribution of genes involved in degradation of plant polymers in this Basidiomycota order, which includes most wood-rotting fungi. Oxidases, including members of the glucose-methanol-choline (GMC) oxidoreductase superfamily, play a central role in the above degradative process because they generate extracellular H2O2 acting as the ultimate oxidizer in both white-rot and brown-rot decay. The survey was completed by analyzing the GMC genes in the available genomes of seven more species to cover the four Polyporales clades. First, an in silico search for sequences encoding members of the aryl-alcohol oxidase, glucose oxidase, methanol oxidase, pyranose oxidase, cellobiose dehydrogenase and pyranose dehydrogenase families was performed. The curated sequences were subjected to an analysis of their evolutionary relationships, followed by estimation of gene duplication/reduction history during fungal evolution. Second, the molecular structures of the near one hundred GMC oxidoreductases identified were modeled to gain insight into their structural variation and expected catalytic properties. In contrast to ligninolytic peroxidases, whose genes are present in all white-rot Polyporales genomes and absent from those of brown-rot species, the H2O2-generating oxidases are widely distributed in both fungal types. This indicates that the GMC oxidases provide H2O2 for both ligninolytic peroxidase activity (in white-rot decay) and Fenton attack on cellulose (in brown-rot decay), after the transition between both decay patterns in Polyporales occurred.

Decolorization of Kraft liquor effluents and biochemical characterization of laccases from Phlebia brevispora BAFC 633

In this work we evaluated the decolorization ability of fungus identified as Phlebia brevispora BAFC 633 on Kraft liquor effluents and we looked at the biochemical characterization of two novel laccases from the fungus. We conducted decolorization assays on diluted Kraft liquor with the enzyme supernatants. Ligninolytic activity was demonstrated by the 20% reduction of lignin peak absorbances between 215 and 280 nm after laccase supernatants treatment.From these supernatants, two novel laccases were isolated and characterized: a constitutively expressed 60-kDa (Lac A), and a CuSO4-induced 75 kDa (Lac B). Laccases were purified by anion-exchange and gel filltration chromatography and their biochemical properties were determined. The temperature and pH optima were 70–75 °C and 4.8 for Lac A, and 60–70 °C and 4.4 for Lac B; and both exhibited high stability at low pH and high temperature. Substrate specificity of the purified enzyme was tested and the highest oxidation was to 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) with a catalytic efficiency of 2.6 × 107 s−1 mol l−1 for Lac A and 9 × 108 s−1 mol l−1 for Lac B.Enzyme supernatants can be used to remediate lignin-rich effluent. The isoenzymes tolerance to low pH and high temperature are significant features to optimize laccase production toward potential biotechnological applications.

Expression of a manganese peroxidase isozyme 2 transgene in the ethanologenic white rot fungus Phlebia sp. strain MG-60

BackgroundThe white-rot fungus Phlebia sp. strain MG-60 was proposed as a candidate for integrated fungal fermentation process (IFFP), which unifies aerobic delignification and semi-aerobic consolidated biological processing by a single microorganism based on its ability to efficiently degrade lignin and ferment the sugars from cellulose. To improve IFFP, the development of a molecular breeding method for strain MG-60 is necessary. The purpose of this study is to establish the transformation method for the strain MG-60 and to obtain the over-expressing transformants of lignin-degrading enzyme, manganese peroxidase.FindingsIn the present study, the expression vector regulated by Phlebia brevispora glyceraldehyde-3-phosphate dehydrogenase promoter and terminator was constructed. A polyethylene glycol transformation method for the ethanol-fermenting white-rot fungus Phlebia sp. MG-60 was established with high transformation efficiency, and the manganese peroxidase isozyme 2 gene (MGmnp2) transformants were obtained, showing higher MnP activity than control transformants. MGmnp2 transformants showed higher selective lignin degradation on Quercus wood powder.ConclusionsThis first report of MG-60 transformation provides a useful methodology for widely accessible to interested researches. These results indicate the possibility of metabolic engineering of strain MG-60 for improving IFFP.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-3-699) contains supplementary material, which is available to authorized users.

Effect of chemical and metallic compounds on biomass, mRNA levels and laccase activity of Phlebia brevispora BAFC 633

Nine aromatic compounds (caffeic acid, syringaldehyde, vanillic acid, guaiacol, vanillin, sinapic acid, syringol, syringic acid and ferulic acid) and four metallic compounds (CuSO4, AgNO3, MnSO4, and CaCl2) were tested for their ability to increase laccase (Lac) activity in the ligninolytic basidiomycete Phlebia brevispora BAFC 633. The addition of syringaldehyde, syringol, guaiacol, sinapic acid, vanillin, ferulic acid and CuSO4 showed a positive effect on fungal growth; however, it decreased dramatically with the addition of AgNO3 and did not undergo changes in the presence of CaCl2 or MnSO4. Lac activity increased with the addition of all the compounds tested, depending on the concentration and the day of culture. P. brevispora BAFC 633 produced two isoenzymes, a constitutively expressed of 60 kDa and another of 75 kDa expressed upon induction by sinapic acid, MnSO4 or CuSO4. Lac secretion capacity of P. brevispora BAFC 633 can be increased 27 times higher than the control with the highest levels detected in the presence of 0.3 mM CuSO4 at day 14. The action is affected at pre-transcriptional level regulating at the onset of the process, however it does not rule out the effect at the post-transcriptional and post-translational levels, for which is necessary to deepen in the knowledge of all possible regulation points of gene expression.

Biopulping of wood chips with Phlebia brevispora BAFC 633 reduces lignin content and improves pulp quality

The white-rot fungus Phlebia brevispora BAFC 633 produces laccases in large proportions. In this work P. brevispora BAFC 633 was grown on Pinus taeda wood chips in 10-L bioreactors. To select the biopulping experimental conditions, we analyzed the variables affecting enzymatic laccase activity in the culture supernatants, indicating that the suitable incubation temperature was 30 °C in order to promote enzyme stability. Phlebia brevispora BAFC 633 secreted 744 U/g of laccase, selectively removing lignin during biotreatment of wood chips, causing a reduction in Kappa number and 10% weight loss, and creating a more open structure and better access to the pulping liquor, which would require less chemical consumption, thus diminishing the environmental impact of the chemical pulping process.These results support the biotechnological potential of P. brevispora BAFC 633 for biopulping processes and enhance the potential for bioprospecting native isolates of the microflora of our country's natural environment.