Lignin degrading system of white-rot fungi and its exploitation for dye decolorization

With global attention and research now focused on looking for the abatement of pollution, white-rot fungi is one of the hopes of the future. The lignin-degrading ability of these fungi have been the focus of attention for many years and have been exploited for a wide array of human benefits. This review highlights the various enzymes produced by white-rot fungi for lignin degradation, namely laccases, peroxidases, aryl alcohol oxidase, glyoxal oxidase, and pyranose oxidase. Also discussed are the various radicals and low molecular weight compounds that are being produced by white-rot fungi and its role in lignin degradation. A brief summary on the developments in research of decolorization of dyes using white-rot fungi has been made.     

<Emphasis Type="Italic">Irpex lacteus</Emphasis>, a white-rot fungus with biotechnological potential — <Emphasis Type="Italic">review</Emphasis>

White-rot fungi that are efficient lignin degraders responsible for its turnover in nature have appeared twice in the center of biotechnological research — first, when the lignin degradation process started being systematically investigated and major enzyme activities and mechanisms involved were described, and second, when the huge remediation potential of these organisms was established. Originally, Phanerochaete chrysosporium became a model organism, characterized by a secondary metabolism regulatory pattern triggered by nutrient (mostly nitrogen) limitation. Last decade brought evidence of more varied regulatory patterns in white-rot fungi when ligninolytic enzymes were also abundantly synthesized under conditions of nitrogen sufficiency. Gradually, research was focused on other species, among them Irpex lacteus showing a remarkable pollutant toxicity resistance and biodegradation efficiency. Systematic research has built up knowledge of biochemistry and biotechnological applicability of this fungus, stressing the need to critically summarize and estimate these scattered data. The review attempts to evaluate the information on I. lacteus focusing on various enzyme activities and bioremediation of organopollutants in water and soil environments, with the aim of mediating this knowledge to a broader microbiological audience.     

Peculiarities of <Emphasis Type="Italic">Pycnoporus</Emphasis> species for applications in biotechnology

The genus Pycnoporus forms a cosmopolitan group of four species belonging to the polyporoid white-rot fungi, the most representative group of homobasidiomycetes causing wood decay. Pycnoporus fungi are listed as food- and cosmetic-grade microorganisms and emerged in the early 1990s as a genus whose biochemistry, biodegradation and biotechnological properties have since been progressively detailed. First highlighted for their original metabolic pathways involved in the functionalization of plant cell wall aromatic compounds to yield high-value molecules, e.g. aromas and antioxidants, the Pycnoporus species were later explored for their potential to produce various enzymes of industrial interest, such as hydrolases and oxidases. However, the most noteworthy feature of the genus Pycnoporus is its ability to overproduce high redox potential laccase—a multi-copper extracellular phenoloxidase—as the predominant ligninolytic enzyme. A major potential use of the Pycnoporus fungi is thus to harness their laccases for various applications such as the bioconversion of agricultural by-products and raw plant materials into valuable products, the biopulping and biobleaching of paper pulp and the biodegradation of organopollutants, xenobiotics and industrial contaminants. All the studies performed in the last decade show the genus Pycnoporus to be a strong contender for white biotechnology. In this review, we describe the properties of Pycnoporus fungi in relation to their biotechnological applications and potential.     

Lignin-modifying enzymes from selected white-rot fungi: production and role from in lignin degradation

Abstract: White-rot fungi produce extracellular lignin-modifying enzymes, the best characterized of which are laccase (EC 1.10.3.2), lignin peroxidases (EC 1.11.1.7) and manganese peroxidases (EC 1.11.1.7). Lignin biodegradation studies have been carried out mostly using the white-rot fungus Phanerochaete chrysosporium which produces multiple isoenzymes of lignin peroxidase and manganese peroxidase but does not produce laccase. Many other white-rot fungi produce laccase in addition to lignin and manganese peroxidases and in varying combinations. Based on the enzyme production patterns of an array of white-rot fungi, three categories of fungi are suggested: (i) lignin-manganese peroxidase group (e.g. P. chrysosporium and Phlebia radiata ), (ii) manganese peroxidase-laccase group (e.g. Dichomitus squalens and Rigidoporus lignosus ), and (iii) lignin peroxidase-laccase group (e.g. Phlebia ochraceofulva and Junghuhnia separabilima ). The most efficient lignin degraders, estimated by ¹⁴CO₂ evolution from ¹⁴C-[Ring]-labelled synthetic lignin (DHP), belong to the first group, whereas many of the most selective lignin-degrading fungi belong to the second, although only moderate to good [¹⁴C]DHP mineralization is obtained using fungi from this group. The lignin peroxidase-laccase fungi only poorly degrade [¹⁴C]DHP.     

白腐菌的研究进展及其在重金属修复中的展望

白腐菌是一类特殊的丝状真菌,能降解多种污染物质,具有广谱、彻底、高效、无专一性的 特点,在生物修复中有广阔的应用前景。综述了白腐菌的分类、酶系、降解机理以及应用于有机 物污染的研究现状,特别介绍了白腐菌在重金属污染的生物修复的应用进展情况,包括白腐菌吸 附重金属的原理、在重金属污染的废水中的研究应用现状及在修复重金属污染土壤中需考虑的 因素。同时展望了白腐菌在重金属污染及复合污染的生物修复中的应用前景。     

White-rot fungi and their enzymes for the treatment of industrial dye effluents

White-rot fungi produce various isoforms of extracellular oxidases including laccase, Mn peroxidase and lignin peroxidase (LiP), which are involved in the degradation of lignin in their natural lignocellulosic substrates. This ligninolytic system of white-rot fungi (WRF) is directly involved in the degradation of various xenobiotic compounds and dyes. This review summarizes the state of the art in the research and prospective use of WRF and their enzymes (lignin-modifying enzymes, LME) for the treatment of industrial effluents, particularly dye containing effluents. The textile industry, by far the most avid user of synthetic dyes, is in need of ecoefficient solutions for its colored effluents. The decolorization and detoxification potential of WRF can be harnessed thanks to emerging knowledge of the physiology of these organisms as well as of the biocatalysis and stability characteristics of their enzymes. This knowledge will need to be transformed into reliable and robust waste treatment processes.     

Mycoremediation (bioremediation with fungi) – growing mushrooms to clean the earth

Abstract

Some of the prospects of using fungi, principally white-rot fungi, for cleaning contaminated land are surveyed. That white-rot fungi are so effective in degrading a wide range of organic molecules is due to their release of extra-cellular lignin-modifying enzymes, with a low substrate-specificity, so they can act upon various molecules that are broadly similar to lignin. The enzymes present in the system employed for degrading lignin include lignin-peroxidase (LiP), manganese peroxidase (MnP), various H₂O₂ producing enzymes and laccase. The degradation can be augmented by adding carbon sources such as sawdust, straw and corn cob at polluted sites.     

Changes in selected enzyme activities during growth of pure and mixed cultures of the white-rot decay fungus Trametes versicolor and the potential biocontrol fungus Trichoderma harzianum.

Two filamentous fungi, the white-rot fungus Trametes versicolor and the soil fungus and potential biocontrol organism Trichoderma harzianum, have been grown in pure and mixed cultures on low-N (0.4 mM) and high-N (4 mM) defined synthetic media to determine the activities of selected wood-degrading enzymes such as cellobiase, cellulase, laccase, and peroxidases. Growth characteristics and enzyme activities were examined for potential correlations. Such correlations would allow the use of simple enzyme assays for measuring biomass development and would facilitate predictions about competitiveness of species in mixed fungal cultures. Our results show that while laccase and Poly Red-478 peroxidase activities indicate survival of the decay fungus, none of the monitored extracellular enzymes can serve as a quantitative indicator for biomass accumulation. As expected, the level of available nitrogen affected the production of the enzymes monitored: in low-N media, specific cellobiase, specific cellulase, and peroxidase activities were enhanced, while laccase activities were reduced. Most importantly, laccase activities of Trametes versicolor, and to a smaller extent, cellobiase activities of both fungi, were significantly induced in mixed cultures of Trametes versicolor and Trichoderma harzianum.     

Biological bleaching of chemical pulps

Use of biotechnology in pulp bleaching has attracted considerable attention and achieved interesting results in recent years. Enzymes of the hemicellulolytic type, particularly xylan-attacking enzymes, xylanases are now used commercially in the mills for pulp treatment and subsequent incorporation into bleach sequences. The aims of the enzymatic treatment depend on the actual mill conditions and may be related to environmental demands, reduction of chemical costs or maintenance or even improvement of product quality. The use of oxidative enzymes from white-rot fungi, that can directly attack lignin, is a second-generation approach, which could produce larger chemical savings than xylanase but has not yet been developed to the full scale. It is being studied in several laboratories in Canada, Japan, the U.S.A. and Europe. Certain white-rot fungi can delignify kraft pulps increasing their brightness and their responsiveness to brightening with chemicals. The fungal treatments are too slow but the enzyme manganese peroxidase and laccase can also delignify pulps and enzymatic processes are likely to be easier to optimize and apply than the fungal treatments. Development work on laccase and manganese peroxidase continues. This article presents an overview of developments in the application of hemicellulase enzymes, lignin-oxidizing enzymes and white-rot fungi in bleaching of chemical pulps. The basic enzymology involved and the present knowledge of the mechanisms of the action of enzymes as well as the practical results and advantages obtained on the laboratory and industrial scale are discussed.     

Impact of white-rot fungi on numbers and community composition of bacteria colonizing beech wood from forest soil

White-rot fungi are important wood-decomposing organisms in forest ecosystems. Their ability to colonize and decompose woody resources may be strongly influenced by wood-inhabiting bacteria that grow on easily utilizable compounds e.g. oligomers of wood-polymers released by fungal enzymes. However, so far, it is not known how white-rot fungi deal with the presence of potential competing bacteria. Here, the effects of two white-rot fungi, Hypholoma fasciculare and Resinicium bicolor, on the numbers and composition of bacteria colonizing sterile beech wood blocks from forest soil are reported. Both total numbers (microscopic counts) and the numbers of cultivable wood-inhabiting bacteria were considerably lower in wood blocks that became colonized by the white-rot fungi than in control blocks. This points to the fungi out-competing the opportunistic bacteria. The presence of white-rot fungi resulted in a change in the relative abundance of families of cultivable bacteria in wood and also in a change of denaturing gradient gel electrophoresis patterns of directly amplified 16S rRNA gene fragments. Analysis of the bacterial community structure in soil adhering to exploratory mycelium (cords) indicated that fungal species-specific effects on bacterial community composition were also apparent in this fungal growth phase.     

Are white-rot fungi a real biotechnological option for the improvement of environmental health?

The use of white-rot fungi as a biotechnological tool for cleaning the environment of recalcitrant pollutants has been under evaluation for several years. However, it is still not possible to find sufficiently detailed investigations of this subject to conclude that these fungi can decontaminate the environment. In the present review, we have summarized and discussed evidence about the potential of white-rot fungi to degrade such pollutants as polycyclic aromatic hydrocarbons, dyes or antibiotics as an example of the complex structures that these microorganisms can attack. This review also discusses field experiment results and limitations of white-rot fungi trials from contaminated sites. Moreover, the use of catabolic potential of white-rot fungi in biopurification systems (biobeds) is also discussed. The current status and future perspectives of white-rot fungi, as a viable biotechnological alternative for improvement of environmental health are noted.     

Decolorization of dyes using white-rot fungi and radical-generating reactions

Decolorization of synthetic dyes was performed using cultures of white-rot fungi producing ligninolytic enzymes and radical-generating reactions that could be involved in the mechanism of fungal decolorization. Among the white-rot fungi tested, Pleurotus ostreatus exhibited the highest decolorization rates, and also the highest production of laccase and Mn-peroxidase. P. ostreatus strain f6 gave 69% decolorization of Eosin Yellowish, 96% of Evans Blue, 75% of Phenol Red (all at 1 mM) and 88% of Poly B-411 (20 ppm) during a 14-day treatment. Treatment with Cu/succinic acid/H₂O₂ resulted in 96% decolorization of Evans Blue and Poly B-411 within 24 h. However, only 48% and 2% decolorization was achieved with Phenol Red and Eosin Yellowish, respectively. Similar decolorization rates were also obtained when Cu was replaced with Co. The results show that treatment of dye-containing solutions with both fungal cultures and biomimetic catalytic reactions results in decolorization.     

Factors affecting mushroom Pleurotus spp.

Pleurotus genus is one of most extensively studied white-rot fungi due to its exceptional ligninolytic properties. It is an edible mushroom and it also has several biological effects, as it contains important bioactive molecules. In basidiomycete fungi, lignocellulolytic enzymes are affected by many typical fermentation factors, such as medium composition, ratio of carbon to nitrogen, pH, temperature, air composition, etc. The survival and multiplication of mushrooms is related to a number of factors, which may act separately or have interactive effects among them. Out that understanding challenges in handling Pleurotus species mushroom requires a fundamental understanding of their physical, chemical, biological and enzymatic properties. This review presents a practical checklist of available intrinsic and extrinsic factors, providing useful synthetic information that may help different users. An in-depth understanding of the technical features is needed for an appropriate and efficient production of Pleurotus spp.     

Biodegradation of environmental pollutants by the white rot fungus Phanerochaete chrysosporium: Involvement of the lignin degrading system

The white-rot fungus Phanrochaete chrysosporium has the ability to degrade a wide variety of structurally diverse organic compounds, including a number of environmentally persistent organopollutants. The unique biodegradative abilities of this fungus appears to be dependent upon its lignin-degrading system. The non-specific and partially extracellular nature of this system suggests that it may be useful as a supplementary means to treat organochemical wastes.     

Distribution of ligninolytic enzymes in selected white-rot fungi

Ten white-rot fungi have been screened for the production of ligninase, manganese peroxidase and laccase. Although the fungi degraded lignin efficiently, they significantly differed in the occurrence of individual ligninolytic enzymes. Based on the enzyme pattern produced under N-limited conditions, the fungi can be divided into the following four groups:1. ligninase-manganese peroxidase-laccase group,2. ligninase-manganese peroxidase group,3. manganese peroxidase-laccase group,4. laccase group.     

White-rots, chlorine and the environment - a tale of many twists

White-rot fungi possess a unique oxidative mechanism by which the recalcitrant lignin component of wood is mineralised. The activity of lignin-degrading enzymes, chiefly lignin and manganese peroxidases, depends on several small organic molecules. Some of these (e.g. chloroanisyl alcohols) are chloroaromatics and may act as environmental pollutants in the forest soil, whereas the synthesis of others (e.g. veratryl alcohol) requires chloromethane. Certain white-rot genera, notably Phellinus and Inonotus, release excess quantities of chloromethane into the atmosphere where it acts as a greenhouse gas. On the other hand, their powerful ligninolytic system enables white-rot fungi to degrade a wide range of man-made environmental pollutants, including recalcitrant chloroaromatics such as DDT, PCP, 2,4-D and 2,4,5-T. This review describes the multifarious interactions of white-rot fungi with their environment via the chlorine cycle.     

Decolorization of dyes using white-rot fungi and radical-generating reactions

Decolorization of synthetic dyes was performed using cultures of white-rot fungi producing ligninolytic enzymes and radical-generating reactions that could be involved in the mechanism of fungal decolorization. Among the white-rot fungi tested, Pleurotus ostreatus exhibited the highest decolorization rates, and also the highest production of laccase and Mn-peroxidase. P. ostreatus strain f6 gave 69% decolorization of Eosin Yellowish, 96% of Evans Blue, 75% of Phenol Red (all at 1 mM) and 88% of Poly B-411 (20 ppm) during a 14-day treatment. Treatment with Cu/succinic acid/H₂O₂ resulted in 96% decolorization of Evans Blue and Poly B-411 within 24 h. However, only 48% and 2% decolorization was achieved with Phenol Red and Eosin Yellowish, respectively. Similar decolorization rates were also obtained when Cu was replaced with Co. The results show that treatment of dye-containing solutions with both fungal cultures and biomimetic catalytic reactions results in decolorization.     

Lignocellulose-degrading marine fungi

Evidence for lignocellulose-degrading ability among marine fungi is reviewed. Enzyme production, mass loss and micromorphological data suggest that most strains capable of decay activity are likely to be soft-rot fungi, with relatively few capable of white-rot decay. This probably reflects the relatively high number of ascomycete genera compared to basidiomycetes described to date. The ecological and biotechnological importance of marine fungal lignocellulolytic enzymes is discussed.     

Utilization and polymerization of lignosulfonates by wood-rotting fungi

The susceptibility of lignosulfonates to the action of lignin-degrading wood-rotting fungi was studied by submitting commercial lignosulfonate (Peritan Na) and fractions of calcium lignosulfonate of different molecular weights to the action of selected white rot fungi. As shown by gel filtration chromatography and determinations according to the nitroso method, lignosulfonates, even in conditions which did not support fungal growth, underwent strong polymerization when brought in contact with typical, extracellular polyphenol oxidase-producing white-rot fungi. Owing to the polymerization, nitroso determinations showed a seeming decrease of lignosulfonate. Polyporus dichrous, an “atypical” white-rot fungus which does not produce extracellular polyphenol oxidase and hence does not cause polymerization of lignosulfonates, was found to degrade 11% of the lignosulfonate available in a solid malt extract medium during 19 days. Addition of lignosulfonate to a rich synthetic liquid growth medium increased the mycelial yield of several white-rot fungi. Trametes versicolor was able to grow on a calcium lignosulfonate fraction with molecular weight 1350 which served as sole source of carbon and energy, but not on fractions of higher molecular weight. The utilization/polymerization of lignosulfonates was shown to depend on concentration and on the presence of additional utilizable sources of carbon.     

A review of biological delignification and detoxification methods for lignocellulosic bioethanol production

Future biorefineries will integrate biomass conversion processes to produce fuels, power, heat and value-added chemicals. Due to its low price and wide distribution, lignocellulosic biomass is expected to play an important role toward this goal. Regarding renewable biofuel production, bioethanol from lignocellulosic feedstocks is considered the most feasible option for fossil fuels replacement since these raw materials do not compete with food or feed crops. In the overall process, lignin, the natural barrier of the lignocellulosic biomass, represents an important limiting factor in biomass digestibility. In order to reduce the recalcitrant structure of lignocellulose, biological pretreatments have been promoted as sustainable and environmentally friendly alternatives to traditional physico-chemical technologies, which are expensive and pollute the environment. These approaches include the use of diverse white-rot fungi and/or ligninolytic enzymes, which disrupt lignin polymers and facilitate the bioconversion of the sugar fraction into ethanol. As there is still no suitable biological pretreatment technology ready to scale up in an industrial context, white-rot fungi and/or ligninolytic enzymes have also been proposed to overcome, in a separated or in situ biodetoxification step, the effect of the inhibitors produced by non-biological pretreatments. The present work reviews the latest studies regarding the application of different microorganisms or enzymes as useful and environmentally friendly delignification and detoxification technologies for lignocellulosic biofuel production. This review also points out the main challenges and possible ways to make these technologies a reality for the bioethanol industry.