Microbial degradation of four biodegradable polymers in soil and compost demonstrating polycaprolactone as an ideal compostable plastic.

Plastics are an indispensable material but also a major environmental pollutant. In contrast, biodegradable polymers have the potential to be compostable. The biodegradation of four polymers as discs, polycaprolactone (PCL), polyhydroxybutyrate (PHB), polylactic acid (PLA) and poly(1,4 butylene) succinate (PBS) was compared in soil and compost over a period of more than 10 months at 25 °C, 37 °C and 50 °C.

Application of green fluorescent protein to measure antimicrobial efficacy and the kinetics of cell death against Escherichia coli.

Industrial antimicrobials have been extensively used to control unwanted microbial growth by incorporation into a variety of products such as plastics and paints, reducing biodeterioration and biofouling and extending the lifespan of the product. Industrial antimicrobials generally have broad sites of action affecting core cellular functions such as central metabolism, enzyme function, cell wall or DNA synthesis and can either be biocidal or biostatic. In addition, susceptibility can be affected by the metabolic state of the microbe, with metabolically inactive cells generally more resistant than metabolically active cells.

Metabarcoding analysis of home composts reveals distinctive fungal communities with a high number of unassigned sequences.

Home composting has been strongly advocated in the UK, Europe and North America to divert organic waste away from conventional waste processing. Despite this, little attention has been given to microbial communities and their diversity in these systems. In this study, we examined the diversity of fungal species in 10 different domestic composts by 454 tag-encoded pyrosequencing.

Prevalence, persistence, and phenotypic variation of Aspergillus fumigatus in the outdoor environment in Manchester, UK, over a 2-year period.

Aspergillus fumigatus, an opportunistic fungal pathogen that causes invasive aspergillosis in immunosuppressed patients, is considered to be the world's most dangerous mould. It is widely distributed in the environment, and airborne asexual conidia serve as the main mode of transport for pulmonary lung infection. It is important to monitor seasonal airborne conidia levels when assessing the risk of acquiring this infection.

Fungal succession in an in-vessel composting system characterized using 454 pyrosequencing.

Fungi are known to have an important role in the composting process as degraders of recalcitrant materials such as cellulose and lignin. Previous attempts to study the diversity and succession of fungi in compost systems have relied on the use of culture-dependent analyses and low-resolution DNA-fingerprinting techniques, lacking the necessary depth to analyse such a rich ecosystem. In this study, 454 pyrosequencing was used to characterize the fungal community composition at the different stages of an in-vessel composting process.

Genetic and virulence variation in an environmental population of the opportunistic pathogen Aspergillus fumigatus.

Environmental populations of the opportunistic pathogen Aspergillus fumigatus have been shown to be genotypically diverse and to contain a range of isolates with varying pathogenic potential. In this study, we combined two RAPD primers to investigate the genetic diversity of environmental isolates from Manchester collected monthly over 1 year alongside Dublin environmental isolates and clinical isolates from patients. RAPD analysis revealed a diverse genotype, but with three major clinical isolate clusters.

Fungal communities associated with the biodegradation of polyester polyurethane buried under compost at different temperatures.

Plastics play an essential role in the modern world due to their low cost and durability. However, accumulation of plastic waste in the environment causes wide-scale pollution with long-lasting effects, making plastic waste management expensive and problematic. Polyurethanes (PUs) are heteropolymers that made up ca.

Laccases of Pleurotus ostreatus observed at different phases of its growth in submerged fermentation: production of a novel laccase isoform.

The production of laccases during the lag, exponential and stationary phases of growth of Pleurotus ostreatus in submerged fermentation was evaluated. Laccase activity was positively correlated to the growth of the fungus. The specific growth rate was 0.

Genomic analysis of the secretion stress response in the enzyme-producing cell factory Aspergillus niger.

Background: Filamentous fungi such as Aspergillus niger have a high capacity secretory system and are therefore widely exploited for the industrial production of native and heterologous proteins. However, in most cases the yields of non-fungal proteins are significantly lower than those obtained for fungal proteins. One well-studied bottleneck appears to be the result of mis-folding of heterologous proteins in the ER during early stages of secretion, with related stress responses in the host, including the unfolded protein response (UPR).

The Aspergillus fumigatus metacaspases CasA and CasB facilitate growth under conditions of endoplasmic reticulum stress.

We have examined the contribution of metacaspases to the growth and stress response of the opportunistic human mould pathogen, Aspergillus fumigatus, based on increasing evidence implicating the yeast metacaspase Yca1p in apoptotic-like programmed cell death. Single metacaspase-deficient mutants were constructed by targeted disruption of each of the two metacaspase genes in A. fumigatus, casA and casB, and a metacaspase-deficient mutant, DeltacasA/DeltacasB, was constructed by disrupting both genes.

Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus.

Aspergillus fumigatus is exceptional among microorganisms in being both a primary and opportunistic pathogen as well as a major allergen. Its conidia production is prolific, and so human respiratory tract exposure is almost constant. A.

Correlation between in vitro growth rate and in vivo virulence in Aspergillus fumigatus.

We describe a kinetic microbroth method of measuring the growth rate of Aspergillus fumigatus spectrophotometrically. Using this method, growth rates were determined for nine A. fumigatus isolates for which an LD90 value in immunosuppressed CD-1 mice had previously been obtained.

Transcriptome analysis of recombinant protein secretion by Aspergillus nidulans and the unfolded-protein response in vivo.

Filamentous fungi have a high capacity for producing large amounts of secreted proteins, a property that has been exploited for commercial production of recombinant proteins. However, the secretory pathway, which is key to the production of extracellular proteins, is rather poorly characterized in filamentous fungi compared to yeast. We report the effects of recombinant protein secretion on gene expression levels in Aspergillus nidulans by directly comparing a bovine chymosin-producing strain with its parental wild-type strain in continuous culture by using expressed sequence tag microarrays.

The Aspergillus niger annexin, anxC3.1 is constitutively expressed and is not essential for protein secretion.

An annexin, anxC3.1, was isolated and characterised from the industrially important filamentous fungus Aspergillus niger. anxC3.

Glutamic protease distribution is limited to filamentous fungi.

Glutamic proteases are a distinct, and recently re-classified, group of peptidases that are thought to be found only in fungi. We have identified and analysed the distribution of over 20 putative glutamic proteases from all fungal species whose genomes have been sequenced so far. Although absent from the Saccharomycetales class, glutamic proteases appear to be present in all other ascomycetes species examined.

Combining transcriptome data with genomic and cDNA sequence alignments to make confident functional assignments for Aspergillus nidulans genes.

Whole genome sequencing of several filamentous ascomycetes is complete or in progress; these species, such as Aspergillus nidulans, are relatives of Saccharomyces cerevisiae. However, their genomes are much larger and their gene structure more complex, with genes often containing multiple introns. Automated annotation programs can quickly identify open reading frames for hypothetical genes, many of which will be conserved across large evolutionary distances, but further information is required to confirm functional assignments.

Oxidative and amphotericin B-mediated cell death in the opportunistic pathogen Aspergillus fumigatus is associated with an apoptotic-like phenotype.

When protoplasts of the opportunistic fungal pathogen Aspergillus fumigatus were treated with low but toxic levels of hydrogen peroxide (0.1 mM) or amphotericin B (0.5 microg ml(-1)), loss of cell viability and death were associated with a number of phenotypic changes characteristic of apoptosis.

Comparison of extracellular phospholipase activities in clinical and environmental Aspergillus fumigatus isolates.

Extracellular phospholipase production by environmental and clinical isolates of Aspergillus fumigatus collected from several centres world-wide were compared. All isolates produced extracellular phospholipases which included phospholipase C and a phospholipid acyl hydrolase (phospholipase A and/or phospholipase B) activity. Clinical isolates of A.

Use of expressed sequence tag analysis and cDNA microarrays of the filamentous fungus Aspergillus nidulans.

The use of microarrays in the analysis of gene expression is becoming widespread for many organisms, including yeast. However, although the genomes of a number of filamentous fungi have been fully or partially sequenced, microarray analysis is still in its infancy in these organisms. Here, we describe the construction and validation of microarrays for the fungus Aspergillus nidulans using PCR products from a 4092 EST conidial germination library.

Entry into the stationary phase is associated with a rapid loss of viability and an apoptotic-like phenotype in the opportunistic pathogen Aspergillus fumigatus.

When the opportunistic pathogen Aspergillus fumigatus entered the stationary phase, there was a rapid loss in cell viability which was associated with the appearance of markers characteristic of apoptosis, namely annexin V-FITC binding to the cytoplasmic membrane, demonstrating exposure of phosphatidylserine to the outer leaflet of the membrane; and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) staining of the nuclei, indicating DNA fragmentation. This was followed later by a loss of membrane integrity as revealed by propidium iodide staining. The development of the apoptotic phenotype was blocked when the protein synthesis inhibitor cycloheximide was added to the culture 1h prior to the onset of the stationary phase, demonstrating active participation of the cell.

Development and application of an assay for uranyl complexation by fungal metabolites, including siderophores.

An assay to detect UO(2)(2+) complexation was developed based on the chrome azurol S (CAS) assay for siderophores (B. Schwyn and J. B.

Glucoamylase::green fluorescent protein fusions to monitor protein secretion in Aspergillus niger.

A glucoamylase::green fluorescent protein fusion (GLA::sGFP) was constructed which allows the green fluorescent protein to be used as an in vivo reporter of protein secretion in Aspergillus niger. Two secretory fusions were designed for secretion of GLA::sGFP which employed slightly different lengths of the glucoamylase protein (GLA499 and GLA514). Expression of GLA::sGFP revealed that fluorescence was localized in the hyphal cell walls and septa, and that fluorescence was most intense at hyphal apices.

Extracellular proteases produced by the Quorn® myco-protein fungus Fusarium graminearum in batch and chemostat culture.

was grown in batch and continuous (chemostat) culture on a glucose-mineral salts medium in the presence and absence of casein. In the absence of casein no protease activity was detected in the culture filtrate from either batch or chemostat culture. For batch cultures grown on medium containing casein, most of the proteolytic activity detected in the supernatant during exponential growth had an optimum at ca pH 5.

How do highly branched (colonial) mutants of Fusarium graminearum A3/5 arise during Quorn myco-protein fermentations?

Chlorate-resistant, highly branched (colonial) mutants and auxotrophic mutants were used to study the nuclear distribution, morphology and growth of heterokaryons of the Quorn myco-protein fungus, Fusarium graminearum A3/5. The results showed that for several complementary homokaryons, even a strong selective pressure was insufficient to maintain heterokaryons in a 'balanced' condition (i.e.

Choline- and acetylcholine-induced changes in the morphology of Fusarium graminearum: evidence for the involvement of the choline transport system and acetylcholinesterase.

The response of Fusarium graminearum to choline, acetylcholine and a number of related analogues was investigated and their ability to induce a morphological response quantified. A number of mutants resistant to the alkylating agent nitrogen mustard (nim strains) were generated and found to have lost the ability to transport choline. These mutants were found to be insensitive to choline and acetylcholine but not to betaine, ethanolamine and other analogues.