Soil fungal communities involved in the biodegradation of polyester polyurethane (PU) were investigated. PU coupons were buried in two sandy loam soils with different levels of organic carbon: one was acidic (pH 5.5), and the other was more neutral (pH 6.7). After 5 months of burial, the fungal communities on the surface of the PU were compared with the native soil communities using culture-based and molecular techniques. Putative PU-degrading fungi were common in both soils, as <45% of the fungal colonies cleared the colloidal PU dispersion Impranil on solid medium. Denaturing gradient gel electrophoresis showed that fungal communities on the PU were less diverse than in the soil, and only a few species in the PU communities were detectable in the soil, indicating that only a small subset of the soil fungal communities colonized the PU. Soil type influenced the composition of the PU fungal communities. Geomyces pannorum and a Phoma sp. were the dominant species recovered by culturing from the PU buried in the acidic and neutral soils, respectively. Both fungi degraded Impranil and represented >80% of cultivable colonies from each plastic. However, PU was highly susceptible to degradation in both soils, losing up to 95% of its tensile strength. Therefore, different fungi are associated with PU degradation in different soils but the physical process is independent of soil type.The worldwide production of synthetic polymers continues to rise, resulting in an increased environmental burden through the generation of plastic waste. More than 140 million tonnes of plastic was produced worldwide in 2001 (34), and the proportion of household plastic waste in the average American home increased from 3 to 5% of total waste in 1969 (15) to more than 30% in 1995 (21) and continues to rise. Many plastics are both physically and chemically robust and cause waste management problems (10). However, several families of plastics undergo biodegradation in the environment, and an understanding of how this degradation occurs may aid in the development of strategies to exploit these processes for waste management purposes.Microorganisms are responsible for the majority of plastic degradation (6), and abiotic factors such as photodegradation or hydrolysis play a very minor role (18,42). Plastics vulnerable to biodegradation include the polyhydroxyalkanoates, polycaprolactone, polylactic acid, polyvinyl chloride (31, 32), and polyester polyurethane (PU). PU is used in a variety of industrial applications, including insulating foams, fibers, and synthetic leather and rubber goods. The presence of ester and urethane linkages in the backbone of PUs makes them susceptible to hydrolysis by enzymes secreted by microorganisms, releasing breakdown products which may act as a carbon source and lead to a weakening of the tensile strength (1,13,22,26,27).Both PU-degrading fungi (5,6,12,32) and bacteria (1, 20, 23) have been isolated from PU, indicating that there are potential reservoirs of PU-degrading organisms widespread in the environment. It...
a b s t r a c tThe identification and annotation of protein-coding genes is one of the primary goals of whole-genome sequencing projects, and the accuracy of predicting the primary protein products of gene expression is vital to the interpretation of the available data and the design of downstream functional applications. Nevertheless, the comprehensive annotation of eukaryotic genomes remains a considerable challenge. Many genomes submitted to public databases, including those of major model organisms, contain significant numbers of wrong and incomplete gene predictions. We present a community-based reannotation of the Aspergillus nidulans genome with the primary goal of increasing the number and quality of protein functional assignments through the careful review of experts in the field of fungal biology.
Multiple Aspergillus fumigatus isolates from a patient with two aspergillomas complicating chronic pulmonary aspergillosis were pan-azole resistant. Microsatellite typing was identical for all isolates despite major phenotypic and some growth rate differences. Three different cyp51A mutations were found (G138C, Y431C, and G434C), of which the first two were demonstrated by heterologous expression in a hypersusceptible Saccharomyces cerevisiae strain to be at least partly responsible for elevated MICs. cyp51A and cyp51B gene duplication was excluded, but increased expression of cyp51A was demonstrated in three isolates selected for additional study (7-to 13-fold increases). In the isolate with the greatest cyp51A expression, an
Background: Fungi can undergo autophagic-or apoptotic-type programmed cell death (PCD) on exposure to antifungal agents, developmental signals, and stress factors. Filamentous fungi can also exhibit a form of cell death called heterokaryon incompatibility (HI) triggered by fusion between two genetically incompatible individuals. With the availability of recently sequenced genomes of Aspergillus fumigatus and several related species, we were able to define putative components of fungi-specific death pathways and the ancestral core apoptotic machinery shared by all fungi and metazoa.
Background: Secretion stress is caused by compromised folding, modification or transport of proteins in the secretory pathway. In fungi, induction of genes in response to secretion stress is mediated mainly by the unfolded protein response (UPR) pathway. This study aims at uncovering transcriptional responses occurring in the filamentous fungi Trichoderma reesei exposed to secretion stress and comparing these to those found in the yeast Saccharomyces cerevisiae.
The aim of this study was to compare biofilm accumulation and heterotrophic bacterial diversity on three pipe materials-cast iron, medium density polyethylene (MDPE), and unplasticised polyvinyl chloride (uPVC) - using a laboratory model system run over a short period (21 d) and a longer period (7 months). Newly Modified Robbins Devices (nMRD) were run in parallel, each containing 25 discs of one material with cold tap water flowing through the devices at 3 ml min(-1) (Reynolds Number 9.05) for 21 d. The numbers of bacteria on each material increased exponentially between 0 and 11 d when the biofilm viable count remained constant. The mean doubling times of the heterotrophic population on the materials during the exponential phase was 13.2 h for cast iron and 15.6 h for MDPE and uPVC. The same experiment was repeated under different environmental conditions with a lower temperature, higher free chlorine and lower number of organisms ml(-1) of incoming water. The exponential phase lengthened to 16 d but the steady state count remained the same. The mean viable count after 21 d and after 7 months was on average 97% higher on cast iron than on the other materials. Very few different colony types were isolated from each material with the largest number (nine) recovered from cast iron. The numbers of planktonic bacteria in the effluent water leaving each of the nMRDs directly correlated with the numbers in the biofilm phase on each of the materials. In addition the distribution and thickness of the biofilms on the MDPE and uPVC were observed using confocal scanning laser microscopy. In conclusion, MDPE and uPVC support the lowest numbers of bacteria in a steady state biofilm in the short term (21 d) and over a longer term (7 months). The diversity of heterotrophic bacteria was greatest on cast iron.
Lactose is the only soluble and economically feasible carbon source for the production of cellulases or heterologous proteins regulated by cellulase expression signals by Hypocrea jecorina (Trichoderma reesei). We investigated the role of the major -galactosidase of H. jecorina in lactose metabolism and cellulase induction. A genomic copy of the bga1 gene was cloned, and this copy encodes a 1,023-amino-acid protein with a 20-amino-acid signal sequence. This protein has a molecular mass of 109.3 kDa, belongs to glycosyl hydrolase family 35, and is the major extracellular -galactosidase during growth on lactose. Its transcript was abundant during growth on L-arabinose and L-arabinitol but was much less common when the organism was grown on lactose, D-galactose, galactitol, D-xylose, and xylitol. ⌬bga1 strains grow more slowly and accumulate less biomass on lactose, but the cellobiohydrolase I and II gene expression and the final cellulase yields were comparable to those of the parental strain. Overexpression of bga1 under the control of the pyruvate kinase promoter reduced the lag phase, increased growth on lactose, and limited transcription of cellobiohydrolases. We detected an additional extracellular -galactosidase activity that was not encoded by bga1 but no intracellular -galactosidase activity. In conclusion, cellulase production on lactose occurs when -galactosidase activity levels are low but decreases as the -galactosidase activities increase. The data indicate that bga1-encoded -galactosidase activity is a critical factor for cellulase production on lactose.The ascomycete Hypocrea jecorina (anamorph, Trichoderma reesei) is used industrially to produce cellulolytic and hemicellulolytic enzymes, and its strong cellulase promoters are of interest for heterologous protein production by this fungus. Cellulase and hemicellulase formation can be induced by several mono-and disaccharides, including sophorose, xylobiose, lactose, D-xylose, and L-sorbose, most of which are too expensive for industrial fermentations. Thus, the range of technically applicable carbon sources is limited. For cellulase and heterologous protein production with cellulase promoters, lactose (1,4-O--D-galactopyranosyl-D-glucose) is virtually the only carbon source that can be used. However, the mechanism by which lactose triggers cellulase formation is not understood. Lactose metabolism is slow, and the cellulase yields on lactose are lower than those on cellulose (27, 28).A key enzyme in the induction of cellulase is galactokinase, since a galactokinase-negative mutant cannot induce cellulases during growth on lactose (30). Lactose can enter the cell after it is hydrolyzed extracellularly to D-glucose and D-galactose by a -galactosidase (lactase, -D-galactoside galactohydrolase; E.C 3.2.1.23), and the monosaccharides are then taken up by the respective permeases. Extracellular -galactosidases are known from several fungi, including Aspergillus niger, Aspergillus oryzae, and Penicillium canescens (5,21,26,29). Alternatively, the...
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