Showing papers by "Andrzej Leonowicz published in 1999"

Immobilization of laccase from Cerrena unicolor on controlled porosity glass

Abstract: White-rot basidiomycete Cerrena unicolor grown in non-induced and induced conditions was tested for production of laccase, lignin peroxidase (LiP) and manganese-dependent peroxidase (MnP). A typical correlation between the concentration of phenolic compounds in the culture fluid and the extracellular laccase activity was observed. The heterogeneous crude laccase preparation obtained after the non-induced fermentor cultivation was immobilized both on controlled porosity glass (CPG) activated by γ-aminopropyltriethoxysilane (APTES) and on CPG with its surface covered by dextran layers. The laccase activities were tested in the aqueous solution for the native and immobilized preparations using different pH and temperature conditions. Laccase activities were additionally examined for native and immobilized forms of laccase preparations in the aqueous solution containing organic solvents. The greatest activity toward the substrate used in the presence of organic solvents was shown by the laccase preparation coupled with the CPG covered by a dextran layer. Potential inhibitors such as thioglycolic acid, thiourea and EDTA used in 1-mM concentration did not show inhibiting properties towards the laccase preparations.

Increased enzymatic activities and levels of superoxide anion and phenolic compounds in cultures of basidiomycetes after temperature stress

Abstract: Selected strains of basidiomycetes (Abortiporus biennis, Trametes versicolor and Cerrena unicolor) were shown to produce enhanced extracellular peroxidase (EP), superoxide dismutase (SOD) and laccase activities following the exposure of 10-day-old fungal cultures to separate high and low temperature stress. The stressful conditions also caused an increase in the concentrations of phenol compounds and superoxide anion radicals in these cultures. At first, peroxidase activity was observed at 12 hours from the moment of temperature stress application. Laccase activity appeared at 96 hours after the maximum levels of superoxide anion radicals (48 h) and SOD activity (36-72 h). The concentration of phenolic substances grew steadily during the period of cultivation. These relations between laccase, SOD and EP as well as superoxide radicals and phenol levels in the environment of ligninolytic fungi seems to be important in the course of the biosynthesis or biodegradation of lignin, as the consequence of adaptation of these basidiomycetes to environmental temperature conditions.

Cooperation of Fungal Laccase and Glucose 1-Oxidase in Transformation of Björkman Lignin and Some Phenolic Compounds

Abstract: A screening of wood-rotting basidiomycete fungi was conducted for glucose 1-oxidase (GOD) and laccase (LAC) production as well as for ligninolytic activity measured by a Rhemazol reaction. The results showed that genera rich in GOD are lignin degraders as well as effective producers of extracellular LAC. The fungi poor in GOD neither showed LAC, nor ligninolytic activity. The Bjorkman lignin and 3 phenolic compounds, hydroquinone and syringic and vanillic acids, were tested on the sequential activity of LAC and GOD. In the presence of LAC, quinoid intermediates formed from Bjorkman lignin and phenolic compounds were observed. The addition of GOD caused a diminution of the quinone level. During incubation of Bjorkman lignin with LAC and GOD depolymerization occurred, and in the experiments omitting GOD the quantities of low molecular products were markedly lower. Consequently, the consecutive ping-pong activity of LAC and GOD reduced the polymerization and improved the efficiency of depolymerization processes.

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

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

Original Articles : The Effects of Wood Rotting Fungi and Laccase on Destaining of Dyes and KP Bleaching Effluent

Abstract: The ability of several wood rotting fungi for decolorization of two anthracene derivatives, Carminic acid (CA) and Remazol brilliant blue R (RBBR), and hardwood KP bleaching liquor (BL) as well as laccase activities in these fungi were studied. The enzyme activity appeared exclusively in fungi destaining RBBR and CA, but in the case of BL, such relationship was not observed. The laccase enzyme was released into the decolorization media and its inducible (but not constitutive) forms shown destaining activity. The purified inducible forms of Kuehneromyces mutabilis and Pleurotus ostreatus laccase destained CA. Thus the possible differentiation between specificity of particular LAC forms was confirmed. In addition the nitrogen starvation induced both laccase and CA destaining activities, but the increase was higher for decolorization of CA than LAC activity. Probably LAC would be only partly responsible for decolorization of this dye. This results suggested that purified LACs decolorize CA, however its destaining activities were considerably lower than the activities on syringaldazine.

The Effects of Wood Rotting Fungi and Laccase on Destaining of Dyes and KP Bleaching Effluent

Abstract: The ability of several wood rotting fungi for decolorization of two anthracene derivatives, Carminic acid (CA) and Remazol brilliant blue R (RBBR), and hardwood KP bleaching liquor (BL) as well as laccase activities in these fungi were studied. The enzyme activity appeared exclusively in fungi destaining RBBR and CA, but in the case of BL, such relationship was not observed. The laccase enzyme was released into the decolorization media and its inducible (but not constitutive) forms shown destaining activity. The purified inducible forms of Kuehneromyces mutabilis and Pleurotus ostreatus laccase destained CA. Thus the possible differentiation between specificity of particular LAC forms was confirmed. In addition the nitrogen starvation induced both laccase and CA destaining activities, but the increase was higher for decolorization of CA than LAC activity. Probably LAC would be only partly responsible for decolorization of this dye. This results suggested that purified LACs decolorize CA, however its destaining activities were considerably lower than the activities on syringaldazine.

The Role of Fungal Laccase in Biodegradation of Lignin

Abstract: Wood components, cellulose and lignin, are degraded simultaneously and the general outline for the complementary character of carbohydrates and lignin decomposition as well as the existence of enzymatic systems combining these processes is still valid The degradatiion of free cellulose or hemicellulose into monosaccharides has long been known to be relatively simple, but the mechanism of lignin degradatiion wasn ot solved very clearly yet Anyway the biodegradation of woold constituents is understood at present as an enzymatic process Kigninolytic activity has been correlated with lignin and manganese peroxidases At present the attention is paid to laccase Laccase oxidizes lignin molecule to phenoxy radicals and quinones This oxidation can lead to the cleavageo f C-C or C-O bonds in the lignin phenyl-propane subunits, resulting either in degradation of both side chains and aromatic rings, or in demethylation processes The role of laccase lies in the "activation" of some low molecular weight mediators and radicals produced by fungal cultures Such activated factors produced also in cooperation with other enzymes are probably exported to the wood environment where they work in degradation processes as the ' enzyme messengers" It is worth mentioning that only fungi possessing laccase show demethylating activity Thus demethylation, the process important for ligninolysis, is probably caused exclusively by laccase Under natural conditions laccase seems to work with other fungal enzymes , mediators and mediating radicals It has shown the possibility of direct Bjrkman lignin depolymerization by cooperative activity of laccase and glucose oxidase

Effect of Superoxide Dismutase and Low Molecular Mediators on Lignin Degradation

Abstract: As the biodegradation of wood constituents has been understood as a multi-basidiomycetes and enzymatic processes, this review will focus on the roles of low molecular compounds and radicals working in harmony with fungal enzymes. Wood rotting basidiomycete fungi penetrate wood, and lead to more easily metabolize carbohydrates of the wood complex. The white-rot fungi, having versatile enzymes, are able to attack directly the "lignin barrier". They also use a multi-enzyme system including so-called "feedback" type enzymes allowing for simultaneous degradation of lignin and carbohydrates. The multi-enzymes including laccase support the proposed route by explaining how the high molecular weight enzymes can function in the wood complex. These enzymes may function separately or cooperate each other. In addition, veratryl alcohol oxidase, cellobiose dehydrogenase, arylalcohol dehydrogenase, and particularly low molecular mediators and radicals have an important role in wood biodegradation. However, the possibility of other mechanism as well as other enzymes, as operating as feedback systems in the process of wood degradation, could not be excluded.