Effects of gadolinium and tin to the production of oxidative enzymes and the growth of five basidiomycetous fungi

Effects of gadolinium (Gd) and tin (Sn) on the growth and production of oxidative enzymes with five basidiomycetous fungi were tested. For this study we have selected well-known white-rot fungi Obba rivulosa and Kuehneromyces mutabilis, in addition to this we have tested three new isolates, the white-rot fungus Phlebia subochracea, the litter-degrading fungus Gymnopus dryophilus and the brown-rot fungus Heliocybe sulcata. This approach allowed us to find possible new sources for oxidative enzymes, such as laccases and versatile peroxidases (VPs). All five tested fungi grew in the presence of Gd (0-200 mg/l) or Sn (0-200 mg/l) on ABTS (2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) containing plates. The growth rate of H. sulcata was tolerant to Gd and Sn (0- 200 mg/l). The growth rates of P. subochracea and G. dryophilus were sensitive to Gd (5-200 mg/l) and Sn (5-200 mg/l). O. rivulosa, K. mutabilis, P. subochracea and G. dryophilus formed colour zones on the ABTS plates indicating that these fungi produced oxidative enzymes, most probably laccases. The brown-rot fungus H. sulcata did not form colour zone on the ABTS plate indicating that this fungus did not produce laccase. The production of laccase with G. dryophilus and K. mutabilis was tolerant to Gd (0-200 mg/l) and Sn (0-200 mg/l). The production of laccase with P. subochracea was sensitive to Gd (5-200 mg/l) and Sn (5-200 mg/l). P. subochracea decolorized the dye Reactive Black 5 without or with Gd and Sn (0- 200 mg/l) indicating the production of VP. O. rivulosa, K. mutabilis, G. dryophilus and H. sulcata did not produce VP. The production of VP by P. subochracea was sensitive to 200 mg/l Gd and Sn.Peer reviewe

Lignin peroxidases, manganese peroxidases, and other ligninolytic enzymes produced by Phlebia radiata during solid-state fermentation of wheat-straw

The white rot fungus Phlebia radiata 79 (ATCC 64658) produces lignin peroxidase (LiP), manganese peroxidase (MnP), glyoxal oxidase (GLOX), and laccase in the commonly used glucose low-nitrogen liquid medium. However, the enzymes which this fungus utilizes for selective removal of lignin during degradation of different lignocellulosic substrates have not been studied before. Multiple forms of LiP, MnP, GLOX, and laccase were purified from P. radiata culture extracts obtained after solid-state fermentation of wheat straw. However, the patterns of extracellular lignin-modifying enzymes studied were different from those of the enzymes usually found in liquid cultures of P. radiata. Three LiP isoforms were purified. The major LiP isoform from solid-state cultivation was LiP2. LiP3, which has usually been described as the major isoenzyme in liquid cultures, was not expressed during straw fermentation. New MnP isoforms have been detected in addition to the previously reported MnPs. GLOX was secreted in rather high amounts simultaneously with LiP during the first 2 weeks of growth. GLOX purified from P. radiata showed multiple forms, with pIs ranging from 4.0 to 4.6 and with a molecular mass of ca. 68 kDa

Secretion of ligninolytic enzymes and mineralization of 14C-ring-labeled synthetic lignin by 3 Phlebia tremellosa strains

Production of ligninolytic enzymes by three strains of the white rot fungus Phlebia tremellosa (syn. Merulius tremellosus) was studied in bioreactor cultivation under nitrogen-limiting conditions. The Mn(II) concentration of the growth medium strongly affected the secretion patterns of lignin peroxidase and laccase. Two major lignin peroxidase isoenzymes were expressed in all strains. In addition, laccase and glyoxal oxidase were purified and characterized in one strain of P. tremellosa. In contrast, manganese peroxidase was not found in fast protein liquid chromatography profiles of extracellular proteins under either low (2.4 µM) or elevated (24 and 120 µM) Mn(II) concentrations. However, H2O2- and Mn-dependent phenol red-oxidizing activity was detected in cultures supplemented with higher Mn(II) levels. Mineralization rates of 14C-ring-labelled synthetic lignin (i.e., dehydrogenation polymerizate) by all strains under a low basal Mn(II) level were similar to those obtained for Phanerochaete chrysosporium and Phlebia radiata. A high manganese concentration repressed the evolution of 14CO2 even when a chelating agent, sodium malonate, was included in the medium

Formation and action of lignin-modifying enzymes in cultures of Phlebia radiata supplemented with veratric acid

Transformation of veratric (3,4-dimethoxybenzoic) acid by the white rot fungus Phlebia radiata was studied to elucidate the role of ligninolytic, reductive, and demeth(ox)ylating enzymes. Under both air and a 100% O2 atmosphere, with nitrogen limitation and glucose as a carbon source, reducing activity resulted in the accumulation of veratryl alcohol in the medium. When the fungus was cultivated under air, veratric acid caused a rapid increase in laccase (benzenediol:oxygen oxidoreductase; EC 1.10.3.2) production, which indicated that veratric acid was first demethylated, thus providing phenolic compounds for laccase. After a rapid decline in laccase activity, elevated lignin peroxidase (ligninase) activity and manganese-dependent peroxidase production were detected simultaneously with extracellular release of methanol. This indicated apparent demethoxylation. When the fungus was cultivated under a continuous 100% O2 flow and in the presence of veratric acid, laccase production was markedly repressed, whereas production of lignin peroxidase and degradation of veratryl compounds were clearly enhanced. In all cultures, the increases in lignin peroxidase titers were directly related to veratryl alcohol accumulation. Evolution of 14CO2 from 3-O14CH3-and 4-O14CH3-labeled veratric acids showed that the position of the methoxyl substituent in the aromatic ring only slightly affected demeth(ox)ylation activity. In both cases, more than 60% of the total 14C was converted to 14CO2 under air in 4 weeks, and oxygen flux increased the degradation rate of the 14C-labeled veratric acids just as it did with unlabeled cultures

Impact of copper and zinc on the growth of saprotrophic fungi and the production of extracellular enzymes

Peer reviewe

Molecular analysis of fungal communities and laccase genes in decomposing litter reveals differences among forest types but no impact of nitrogen deposition

The fungal community of the forest floor was examined as the cause of previously reported increases in soil organic matter due to experimental N deposition in ecosystems producing predominantly high-lignin litter, and the opposite response in ecosystems producing low-lignin litter. The mechanism proposed to explain this phenomenon was that white-rot basidiomycetes are more important in the degradation of high-lignin litter than of low-lignin litter, and that their activity is suppressed by N deposition. We found that forest floor mass in the low-lignin sugar-maple dominated system decreased in October due to experimental N deposition, whereas forest floor mass of high-lignin oak-dominated ecosystems was unaffected by N deposition. Increased relative abundance of basidiomycetes in high-lignin forest floor was confirmed by denaturing gradient gel electrophoresis (DGGE) and sequencing. Abundance of basidiomycete laccase genes, encoding an enzyme used by white-rot basidiomycetes in the degradation of lignin, was 5–10 times greater in high-lignin forest floor than in low-lignin forest floor. While the differences between the fungal communities in different ecosystems were consistent with the proposed mechanism, no significant effects of N deposition were detected on DGGE profiles, laccase gene abundance, laccase length heterogeneity profiles, or phenol oxidase activity. Our observations indicate that the previously detected accumulation of soil organic matter in the high-lignin system may be driven by effects of N deposition on organisms in the mineral soil, rather than on organisms residing in the forest floor. However, studies of in situ gene expression and temporal and spatial variability within forest floor communities will be necessary to further relate the ecosystem dynamics of organic carbon to microbial communities and atmospheric N deposition.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72825/1/j.1462-2920.2007.01250.x.pd