Maximum Manganese-dependent Peroxidase Research Articles

Lignin in Paper Mill Sludge is degraded by White-Rot Fungi in Submerged Fermentation

This study investigated biological treatment of paper mill sludge and lignin degradation by fungi. Four different Basidiomycetes white-rot fungi (WRF) were used: Coriolus versicolor, Tyromyces albidus, Trametes gallica and Pleurotus ostreatus. The fungi were cultured in submerged fermentation cultures with paper mill sludge. The pH values of the cultures and ligninolytic enzymes production profiles of the four fungi were monitored and reported here. The highest laccase activity of 202 U/L was obtained in T. albidus at day 25 while the maximum manganese-dependent peroxidase (MnP) activity of 50 U/L was obtained in C. versicolor from day 15 to 25. The P. ostreatus showed highest total cellulase activity with the peak of 0.26 μmol at day 15 while the other three WRF showed negligible total cellulase activity. The lignin contents were significantly decreased in the four WRF-treated sludge samples and were confirmed by acetyl bromide and FTIR analyses. The results suggested that lignin was preferentially used and significantly degraded or converted in all the four WRF-treated sludge samples.

Biodegradation potential and ligninolytic enzyme activity of two locally isolated Panus tigrinus strains on selected agro-industrial wastes

The degradation potential and ligninolytic enzyme production of two isolated Panus tigrinus strains (M609RQY and M109RQY) were evaluated in this study. These strains were grown on three selected abundant agro-industrial wastes (rice straw; rice husk and cassava peel) under solid-state fermentation conditions. Degradation potential was determined by analyzing the chemical composition of the selected substrates before and after fermentation along with ligninolytic enzyme production. The strain M609RQY led to the highest lignin degradation of 40.81% on cassava peel, 11.25% on rice husk and 67.96% on rice straw. Both strains significantly increased the protein content of cassava peel. Rice husk stimulated maximum laccase (2556 U/L) and lignin peroxidase (24 U/L) production by the strains M109RQY and M609RQY, respectively. Furthermore, cassava peel stimulated maximum manganese-dependent peroxidase (141 U/L) production by the strain M109RQY. The de-lignified rice straw and the nutritionally-improved cassava peel could serve as potential animal feed supplements.

Manganese Peroxidase Production by Immobilized Phanerochaete chrysosporium BKM‐F‐1767 in a Cell Bioreactor

Manganese‐dependent peroxidase (MnP) production was performed in an immobilized cell bioreactor in which Phanerochaete chrysosporium BKM‐F‐1767 was immobilized on polystyrene foam. The immobilized cell culture yielded significantly greater MnP activity than the conventional stationary liquid culture. Cultivation was carried out in batch mode; the effect of glucose concentration was investigated and growth kinetics parameters were found as, μmax=0.59 day−1, K s =0.33 g/L and K ss =14.5. Batch operation led to maximum MnP (770.82 U/L) in the culture medium containing 0.05% Tween 80, 10 g/L glucose, and 174 µM Mn2+ at 37°C and pH 4.5. Enzyme productivity was obtained as 110.12 U/day/L.

Enhanced production of manganese peroxidase by Phanerochaete chrysosporium

Production of manganese-dependent peroxidase (MnP) by the white-rot fungus Phanerochaete chrysosporium BKM-F-1767 (ATCC 24725) was monitored during growth in different media and growth conditions. The effect of some activators of MnP production, Mn2+, Tween 80, phenylmethylsulphonylfloride (PMSF), oxygen, temperature, pH, glycerol and nitrogen was studied. Supplementing the cultures with Tween 80 (0.05 %, v/v) and Mn2+ (174 µM) resulted a maximum MnP activity of 356 U/L which was approximately two times higher than that obtained in the control culture (without Tween 80). Decolourisation of Direct Blue 15 and Direct Green 6 (50 mg/L) was also achieved with MnP.

Production and stimulation of manganese peroxidase by immobilized Phanerochaete chrysosporium

The effect of some activators of manganese-dependent peroxidase (MnP) production, Mn 2+, Tween 80, phenylmethylsulphonylfloride (PMSF), oxygen, temperature, pH, and glycerol and nitrogen amount to solid-state cultures of Phanerochaete chrysosporium BKM-F-1767 (ATCC 24725) was studied. Polystyrene foam beads were employed as support material. Supplementing the cultures with Tween 80 (0.05%, v/v) and Mn 2+ (174 μM) a maximum MnP activity of 421 U/l was achieved. This value was approximately two times higher than that obtained in the control cultures (without Tween 80). Decolorization of some azo dyes was also achieved with MnP that produced in solid-state culture of P. chrysosporium.

Production of laccase and manganese peroxidase by Fomes sclerodermeus grown on wheat bran.

The aim of this work was to study the growth and production of ligninolytic enzymes by Fomes sclerodermeus using a natural medium based on wheat bran as the principal substrate in a solid-state fermentation. Growth was monitored by measuring the chitin content in the substrate. The maximum rate of growth was observed between days 7 and 18. A 38% total dry-weight loss of the substrate was measured after 28 days of cultivation. Differential hydrolysis of the substrate revealed that cellulose was more extensively degraded than lignin. In the 28-day incubation period, the losses of cellulose and lignin were 38 and 15%, respectively. No lignin peroxidase activity was found in any of the media tested. The maximum manganese-dependent peroxidase activity recorded was 6.3 U g(-1) at 14 days, while the maximum laccase activity was 270 U g(-1) at 28 days post-inoculation. Addition of commonly used inducers such as copper or manganese did not produce a further increase in the enzyme activities, nor did addition of glucose, asparagine, or malt extract.

Production of manganese-dependent peroxidase in a new solid-state bioreactor by Phanerochaete chrysosporium grown on wood shavings. Application to the decolorization of synthetic dyes.

The production of manganese-dependent peroxidase (MnP) by Phanerochaete chrysosporium in a new solid-state bioreactor, the immersion bioreactor, operating with lignocellulosic waste, such as wood shavings, was investigated. Maximum MnP and lignin peroxidase (LiP) activity of 13.4 and 8.48 mukat/L were obtained, respectively. The in vitro decolorization of several synthetic dyes by the extracellular liquid produced in the above-mentioned bioreactor (containing mainly MnP) was carried out and its degrading ability was assessed. The highest decolorization was reached with Indigo Carmine (98%) followed by Bromophenol Blue (56%) and Methyl Orange (36%), whereas Gentian Violet was hardly decolorized (6%).

Performance of a solid-state immersion bioreactor for ligninolytic enzyme production: evaluation of different operational variables

The production of ligninolytic enzymes by the white-rot fungus Phanerochaete chrysosporium BKM-F-1767 in a solid-state immersion bioreactor, employing cubes of nylon sponge as a support, was studied. Cultivation was carried out in both batch and continuous mode, and the effect of some operational variables (aeration level, pH) was investigated. Batch operation at an aeration level of 0.5 vvm led to maximum manganese-dependent peroxidase (MnP) and lignin peroxidase (LiP) activities of 574 and 116 U l −1, respectively. The results were compared with those obtained at a higher aeration level (1 vvm), reported in a previous work, and it appeared that LiP productivity increased with aeration rate, while MnP was not significantly affected. Continuous operation showed much lower MnP activities (around 60 U l −1) and similar LiP activities (about 132 U l −1). However, in terms of productivity the difference between batch and continuous operation for MnP was less remarkable (239 and 150 U day −1, respectively), whereas LiP productivity was ten-fold higher in continuous operation than in batch mode. This could be attributed to the influence of operation pH on ligninolytic enzyme activities. The study of the kinetic characteristics of the biocatalysts supported this hypothesis.

Design of a new rotating drum bioreactor for ligninolytic enzyme production by Phanerochaete chrysosporium grown on an inert support

The production of ligninolytic enzymes by the white-rot fungus Phanerochaete chrysosporium BKM-F-1767 was studied in a new bioreactor configuration based on a standard rotating drum bioreactor. P. chrysosporium was grown on cubes of nylon sponge, and cultivation was carried out in batch. Two aeration levels: 0.5 and 1 vvm were tested. The latter led to activities about 3-fold higher than the former, achieving maximum manganese-dependent peroxidase (MnP) and lignin peroxidase (LiP) activities of 1350 and 364 U/l, respectively. Moreover, laccase activity was also detected, showing a highest activity of 56 U/l. In addition, the in vitro decolorisation of a model dye (Poly R-478) by the extracellular liquid obtained in the bioreactor was monitored in order to assess its ligninolytic ability. A percentage of Poly R-478 decolorisation of about 19% was achieved, after 15 min of dye incubation.

Influence of Several Activators on the Extracellular Laccase Activity andIn vivo Decolourization of Poly R-478 by Semi-Solid-State Cultures ofTrametes versicolor

The effect of several laccase activity activators,such as ethanol (novel activator), veratryl alcohol, melanin production and aeration level, on the laccase production by Trametes versicolor (CBS100.29) was investigated. The microorganism was cultivated on nylon sponge, functioning as a physical support on which the mycelium was bound. The cultures with veratryl alcohol showed maximum laccase and manganese-dependent peroxidase (MnP) activities of 238 U/l and 125 U/l, respectively. The laccase activity found is about two times higher than that attained in the control cultures. On the contrary, MnP activity did not appear to be influenced by the addition of this alcohol. Ethanol-supplemented cultures led to maximum laccase and MnP activity levels of about 102 U/l and 101 U/l, respectively. These activities were approx. 40% lower than those achieved in the reference cultures. The decolourization of the polymeric dye Poly R-478 by the above-mentioned cultures was also investigated. A percentage of biological decolourization of around 90% was achieved with control and veratryl alcohol-supplemented cultures, whereas with ethanol-supplemented cultures a slightly lower percentage of around 85% was reached after seven days of dye incubation.

Design of different bioreactor configurations: application to ligninolytic enzyme production in semi-solid-state cultivation

The production of ligninolytic enzymes by Phanerochaete chrysosporium BKM-F-1767 (ATCC 24725) in laboratory-scale bioreactors was studied. The cultivations were carried out in semi-solid-state conditions, employing corncob as carrier, which functioned both as a place of attachment and as a source of nutrients. Several bioreactor configurations were investigated in order to determine the most suitable one for ligninolytic enzyme production: a 1-dm3-static-bed bioreactor, a 1-dm3-static-bed bioreactor with air diffusers into the bed, a 0.5-dm3-static-bed bioreactor with air diffusers into the bed and a tray bioreactor. Although the static-bed configurations produced maximum individual lignin peroxidase (LiP) activities about 400 U dm−3 (1.0-dm3 bioreactor) and about 700 U dm−3 (0.5-dm3 bioreactor), manganese-dependent peroxidase (MnP) was not detected throughout the cultures. Nevertheless, the tray configuration led to maximum individual MnP and LiP activities of about 200 U dm−3 and 300 U dm−3, respectively. Therefore, this configuration is the most adequate of the different bioreactor configurations tested in the present work, since the ligninolytic complex formed by MnP and LiP is more efficient for its application to bio-processing systems. In addition, the results indicated the influence of the oxygen in ligninolytic enzyme production. © 2001 Society of Chemical Industry

Stimulation of ligninolytic enzyme production and the ability to decolourise Poly R-478 in semi-solid-state cultures of Phanerochaete chrysosporium

The effect of adding some activators of ligninolytic enzyme production, Tween 80, veratryl alcohol and manganese (IV) oxide, to semi-solid-state cultures of Phanerochaete chrysosporium BKM-F-1767 (ATCC 24725) was studied. As carrier, cubes of polyurethane foam were employed. Supplementing the cultures with Tween 80, a maximum manganese-dependent peroxidase (MnP) activity of 350 U/l was achieved. This value was about 3-fold higher than that obtained in the control cultures (without Tween 80). When not only Tween 80 but also veratryl alcohol or manganese (IV) oxide were incorporated into the culture medium, the effect was greater and led to MnP activities of about 600 and 1100 U/l with alcohol or oxide, respectively. These activities were notably higher than those obtained in the control cultures (about 3-fold and 7-fold, respectively). The decolourisation of the dye Poly R-478 by the above-mentioned cultures was investigated. The percentage of biological decolourisation reached at the end of all the cultivations was higher than 95%.

Extracellular ligninolytic enzyme production by Phanerochaete chrysosporium in a new solid-state bioreactor

The production of manganese-dependent peroxidase (MnP) and lignin peroxidase (LiP) by the fungus Phanerochaete chrysosporium (ATCC 24725) in a new bioreactor, the Immersion Bioreactor, which grows cells under solid-state conditions, was studied. Maximum MnP and LiP activities were 987 U l−1 and 356 U l−1, respectively. The polymeric dye, Poly R-478, was degraded at 2.4 mg l−1 min−1 using the extracellular culture filtrate.

In vivo decolourization of the polymeric dye poly R‐478 by corncob cultures of Phanerochaete chrysosporium

AbstractIn this paper, the in vivo decolourization of the polymeric dye Poly R‐478 by semi‐solid‐state cultures of Phanerochaete chrysosporium BKM‐F‐1767 (ATCC 24725) was investigated, employing corncob as a support. In order to stimulate the ligninolytic system of the fungus, the cultures were supplemented with veratryl alcohol (2 mM) or manganese (IV) oxide (1 g/l).Maximum manganese‐dependent peroxidase (MnP) and lignin peroxidase (LiP) activities of around 2,000 U/l and 400 U/l were attained by the former, whereas the activities reached by the latter were of about 1,500 U/l and 200 U/l, respectively. Furthermore, laccase activity (around 150 U/l) was only detected in manganese (IV) oxide supplemented cultures.The polymeric dye Poly R‐478 (0.02 w/v) was added to three‐day‐old cultures. A percentage of biological decolourization of about 85% was achieved using cultures supplemented with veratryl alcohol, whereas MnO2 cultures showed a rather lower percentage of around 58% after nine days of dye incubation. Moreover, a correlation between MnP activity and Poly R‐478 decolourization could be observed, indicating that this enzyme is mainly responsible for dye degradation.In the present work, the in vivo decolourizing capability of the ligninolytic complex secreted by P. chrysosporium was investigated under the above‐mentioned cultivation conditions, employing a model compound, such as the polymeric dye Poly R‐478.