Penicillium Solitum Research Articles

Gut microeukaryotes during anorexia nervosa: a case report

BackgroundFew studies have focused on eukaryote community in the human gut. Here, the diversity of microeukaryotes in the gut microbiota of an anorexic patient was investigated using molecular and culture approaches.Case presentationA 21-year-old Caucasian woman was admitted in an intensive care unit for severe malnutrition in anorexia nervosa. One stool specimen was collected from the anorexic patient, culture and polymerase chain reaction-based explorations yielded a restricted diversity of fungi but four microeukaryotes Tetratrichomonas sp., Aspergillus ruber, Penicillium solitum and Cladosporium bruhnei previously undescribed in the human gut.ConclusionsEstablishing microeukaryote repertoire in gut microbiota contributes to the understanding of its role in human health.

Attraction of Walnut Twig Beetle Pityophthorus juglandis (Coleoptera: Curculionidae) to the Fungus Geosmithia morbida

Geosmithia morbida causes thousand cankers disease of Juglans nigra and it is transmitted by the walnut twig beetle (WTB, Pityophthorus juglandis). Along with WTB, an ambrosia beetle, Xyleborinus saxeseni, is commonly associated with thousand cankers disease in the later stages of J. nigra decline, although X. saxeseni is not a known vector of G. morbida. We initiated studies to determine whether WTBs or X. saxeseni were attracted to volatiles produced by G. morbida and other bark fungi in a laboratory choice-test setting. There was no difference between the numbers of emerged WTB adults that were collected in tubes containing potato dextrose agar colonized by G. morbida and those in tubes containing agar only. More adult X. saxeseni were collected in tubes containing agar only compared with G. morbida. Walnut twig beetle larvae migrated more frequently toward an agar plug colonized by G. morbida and Fusarium solani compared with an un-colonized agar plug. No larval preference was observed when agar plugs colonized by F. solani and G. morbida, or G. morbida and Penicillium solitum, were placed in the same petri dish. These results suggest that WTB larvae are attracted to bark fungi in general, but not specifically to G. morbida. Accepted 27 June 2014. Published 13 August 2014.

An Evaluation of the Proteolytic and Lipolytic Potential of Penicillium spp. Isolated from Traditional Greek Sausages in Submerged Fermentation

A number of novel Penicillium strains belonging to Penicillium nalgiovense, Penicillium solitum, Penicillium commune, Penicillium olsonii, and Penicillium oxalicum species, isolated from the surface of traditional Greek sausages, were evaluated for their proteolytic and lipolytic potential in a solid substrate first and next in submerged fermentations, using complex media. Extracellular proteolytic activity was assessed at acid, neutral, and alkaline pH, while the lipolytic activity was assessed using olive oil, the short-chain triacylglycerol tributyrin, and the long-chain triolein, as substrates. The study revealed that although closely related, the tested strains produce enzymes of distinct specificities. P. nalgiovense PNA9 produced the highest alkaline proteolytic activity (13.2 unit (U)/ml) and the highest lipolytic activity with tributyrin (92 U/ml). Comparisons with known sources show that proteases and/or lipases can be secreted effectively by some Penicillia (P. nalgiovense PNA4, PNA7, and PNA9 and P. solitum PSO1), and further investigations on their properties and characteristics would be promising.

Penicillium solitum: a mesophilic, psychrotolerant fungus present in marine sediments from Antarctica

Few studies have addressed the diversity of cultivable fungi from marine sediments, especially those from Antarctica. In the present study, we evaluated the presence and distribution of cultivable fungi in marine core sediments obtained from 100, 500, 700 and 1,100 m below the Antarctic Ocean surface. Fifty-two fungal isolates were identified as Penicillium solitum by their physiological and morphological characteristics, and the identity of 12 representative isolates was further confirmed by sequencing of the ITS1-5.8S-ITS2 and β-tubulin genes. P. solitum displayed high sequence similarity to Penicillium taxa that have been described from other marine habitats. Conidial germination of P. solitum occurred at low temperatures and high salinities. In addition, P. solitum displayed extracellular amylasic and esterasic activities. The isolation of P. solitum from marine sediments in Antarctica and its survival at low temperatures and high salt concentrations suggest that it is adapted to the cold and halophilic environment of the Antarctic oceans. Because P. solitum produces extracellular enzymes, it is an interesting eukaryotic model for the study of structure–function relationships during enzymatic biocatalysis and biotransformation under extreme conditions. Marine sediments from Antarctica may represent a unique source for obtaining extremophilic fungi. New studies using different culture media, temperatures ranges and pressure conditions as well as metagenomic techniques can assist in understanding the extremophilic fungal communities in marine sediments across the Antarctic Ocean.

Ectomycota Associated with Hibernating Bats in Eastern Canadian Caves prior to the Emergence of White-Nose Syndrome

Abstract - The emergence of the fungal disease white-nose syndrome (WNS) among hibernating bats in North America and its causative pathogen, Geomyces destructans, underscores how little is known about fungi associated with bats and their subterranean environments. Investigating 8 caves and mines in New Brunswick, Canada, we cultured a diverse array of fungi from the fur and skin of apparently healthy, hibernating Myotis lucifugus (Little Brown Bat) and M. septentrionalis (Northern Long-eared Bat) in the year prior to the emergence of WNS in the province. Among the 117 isolated fungal taxa, we found an array of psychrophilic, psychrotrophic, keratinolytic, coprophilous, and saprobic fungi. The most common taxa were Geomyces pannorum sensu lato, Penicillium spp., Mortierella spp., Mucor spp., Cephalotrichum stemonitis, Leuconeurospora spp., Penicillium solitum, Cladosporium spp., and Trichosporon dulcitum. Each bat hosted 6.9 ± 3 (SD) fungal taxa, and 30.8 ± 5 taxa were isolated per hibernaculum. Number of ...

Penicillium solitum produces a polygalacturonase isozyme in decayed Anjou pear fruit capable of macerating host tissue in vitro

A polygalacturonase (PG) isozyme was isolated from Penicillium solitum-decayed Anjou pear fruit and purified to homogeneity with a multistep process. Both gel filtration and cation exchange chromatography revealed a single PG activity peak, and analysis of the purified protein showed a single band with a molecular mass of 43 kDa, which is of fungal origin. The purified enzyme was active from pH 3.5–6, with an optimum at pH 4.5. PG activity was detectable 0–70 C with 50 C maximum. The purified isozyme was inhibited by the divalent cations Ca2+, Mg2+, Mn2+ and Fe2+ and analysis of enzymatic hydrolysis products revealed polygalacturonic acid monomers and oligomers. The purified enzyme has an isoelectric point of 5.3 and is not associated with a glycosylated protein. The PG isozyme macerated fruit tissue plugs in vitro and produced ~1.2-fold more soluble polyuronides from pear than from apple tissue, which further substantiates the role of PG in postharvest decay. Data from this study show for the first time that the purified PG produced in decayed Anjou pear by P. solitum, a weakly virulent fungus, is different from that PG produced by the same fungus in decayed apple.

Complete mitochondrial genome of compactin-producing fungus Penicillium solitum and comparative analysis of Trichocomaceae mitochondrial genomes

We determined the complete mitochondrial genome sequence of the compactin-producing fungus Penicillium solitum strain 20-01. The 28601-base pair circular-mapping DNA molecule encodes a characteristic set of mitochondrial proteins and RNA genes and is intron-free. All 46 protein- and RNA-encoding genes are located on one strand and apparently transcribed in one direction. Comparative analysis of this mtDNA and previously sequenced but unannotated mitochondrial genomes of several medically and industrially important species of the Aspergillus/Penicillium group revealed their extensive similarity in terms of size, gene content and sequence, which is also reflected in the almost perfect conservation of mitochondrial gene order in Penicillium and Aspergillus. Phylogenetic analysis based on concatenated mitochondrial protein sequences confirmed the monophyletic origin of Eurotiomycetes.

Caspase-1 and -3 Inhibiting Drimane Sesquiterpenoids from the Extremophilic Fungus Penicillium solitum

Two new drimane sesquiterpene lactones and one new tricarboxylic acid derivative were isolated from the Berkeley Pit extremophilic fungus Penicillium solitum. The structures of these compounds were deduced by spectroscopic analysis. Berkedrimanes A and B inhibited the signal transduction enzymes caspase-1 and caspase-3 and mitigated the production of interleukin 1-β in the induced THP-1 (pro-monocytic leukemia cell line) assay.

Temperature Suppresses Decay on Apple Fruit by Affecting Penicillium solitum Conidial Germination, Mycelial Growth and Polygalacturonase Activity

Temperature Suppresses Decay on Apple Fruit by Affecting Penicillium solitum Conidial Germination, Mycelial Growth and Polygalacturonase Activity

Isolation, Purification, and Characterization of a Polygalacturonase Produced in Penicillium solitum-Decayed ‘Golden Delicious’ Apple Fruit

Polygalacturonase (PG) was extracted and purified from decayed 'Golden Delicious' apple fruit inoculated with Penicillium solitum. Ammonium sulfate, gel filtration, and cation exchange chromatography were used to purify the enzyme. Both chromatographic methods revealed a single peak corresponding to PG activity. The purified PG most likely originates from the fungus because PG activity from healthy and wounded apple tissue was undetectable. Analysis of cation exchange-purified material using sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a single 50-kDa band. The enzyme was active over a broad pH range (3 to 7), with optimal activity between pH 4 and 5. PG was highly active at 20 and 37 degrees C but was also detectable at 2, 50, and 75 degrees C. Divalent cations affected PG enzyme activity; Mg and Fe increased, whereas Ca and Mn reduced activity in vitro. Thin-layer chromatographic separation of hydrolysis products and data from a PG plate activity assay based on staining with ruthenium red showed that the enzyme exhibits both exo and endo activity. Purified PG incubated with intact apple fruit tissue in vitro caused a 30% reduction in mass after 48 h, suggesting a role in P. solitum-mediated decay of apple fruit.

Improved monoterpene biotransformation with Penicillium sp. by use of a closed gas loop bioreactor

A closed gas loop bioprocess was developed to improve fungal biotransformation of monoterpenes. By circulating monoterpene-saturated process gas, the evaporative loss of the volatile precursor from the medium during the biotransformation was avoided. Penicillium solitum, isolated from kiwi, turned out to be highly tolerant towards monoterpenes and to convert alpha-pinene to a range of products including verbenone, a valuable aroma compound. The gas loop was mandatory to reproduce the production of 35 mg L(-1) verbenone obtained in shake flasks and also in the bioreactor. Penicillium digitatum DSM 62840 regioselectively converted (+)-limonene to the aroma compound alpha-terpineol, but shake flask cultures revealed a pronounced growth inhibition when initial concentrations exceeded 1.9 mM. In the bioreactor, toxic effects on P. digitatum during biotransformation were alleviated by starting a sequential feeding of non-toxic limonene portions after a preceding growth phase. Closing the precursor-saturated gas loop during the biotransformation allowed for an additional replenishment of limonene via the gas phase. The gas loop system led to a maximum alpha-terpineol concentration of 1,009 mg L(-1) and an average productivity of 8-9 mg L(-1) h(-1) which represents a doubling of the respective values previously reported. Furthermore, a molar conversion yield of up to 63% was achieved.

Enzymatic hydrolysis of salmon oil by native lipases: optimization of process parameters

In an attempt to concentrate the content of n-3 polyunsaturated fatty acids (n-3 PUFA) in the residual acylglycerol, salmon oil (n-3 PUFA content of 30.1%) was hydrolyzed with three kinds of native microbial lipases (Aspergillus niger, Rhizopus javanicus and Penicillium solitum). For each lipase, a response surface methodology was used to obtain maximum PUFA content and to optimize the parameters of enzymatic reactions with respect to important reaction variables; temperature (X1), amount of lipases (X2) and water/oil ratio (X3). Based on these results, optimal reaction conditions were established. Aspergillus niger lipase was the most effective in concentrating n-3 PUFA. The degree of hydrolysis (60%) led to an increase in the docosahexaenoic acid (DHA) content from 14.4% in the original oil to 34.0% (2.4-fold enrichment) in the residual acylglycerol under optimum conditions: enzyme concentration of 500 U g-1 oil, reaction temperature of 45 ºC and water/oil mass rate of 2:1 (m/m) after 24 h reaction.

Moulds contaminants on Norwegian dry-cured meat products

Dry-cured meat production has a long tradition in Norway. However, uncontrolled mould growth on the surface of the dry-cured meat products is causing significant quality problems. As some moulds are mycotoxigenic, their growth on the dry-cured meat products could also pose a serious health risk. Such quality problems and potential health risks can be better handled if the types of moulds growing on the products are known. In total, 161 samples were collected from the ripening and packaging stages of production with the aim of identifying moulds contaminating smoked and unsmoked Norwegian dry-cured meat products. Moulds were isolated either by transferring aerial mycelium of each visible mould colonies on the products or by directly plating pieces of meat on suitable agar media. The isolates were identified at a species level by a polyphasic approach. In total, 264 isolates belonging to 20 species and four fungal genera were identified. The genus Penicillium covered 88.3% of the total isolates. This genus contributed to the isolates of smoked and unsmoked products by 91% and 84% respectively. Penicillium nalgiovense was the dominant species isolated from both smoked and unsmoked products and covered 38% of the total isolates. Penicillium solitum and P. commune were the next most frequently isolated species with a contribution of 13% and 10% respectively. Species of Cladosporium and Eurotium contributed to the total isolates by 6% and 4.9% respectively. Smoking seems to affect the growth of these dominating species differently. An increase in the isolation frequency of P. nalgiovense accompanied by the reduction in the occurrence of P. solitum, P. commune and species of Cladosporium was observed on smoked products. The survey showed that the species of Penicillium are associated with Norwegian dry-cured meat products in general. Penicillium nalgiovense, the dominating mould species, is a potential producer of penicillin and its presence could represent a threat to allergic consumers.

Immobilization of Penicillium solitum lipase on the carbon fiber material “Dnepr-MN”

A method for immobilizing Penicillium solitum lipase (the medicinal agent Solizyme) on the activated carbon fiber material “Dnepr-MN” is proposed; using a matrix with total pore volumes of 0.41–1.3 cm3/g, the method allows the enzyme:matrix ratio (1:30–1:60) to be controlled, yielding preparations with a specified lipolytic activity (20–40 LU/mg of agent) at quantitative yield (91–97%). Studies of the pH and temperature stability, functioning in acid media, and desorption demonstrated stabilization and prolongation of the lipolytic activity of the immobilized lipase.

In Vivo ESR Spin Trapping Detection of Carbon-Centered α-Farnesene Radicals

Lyophilisates of the ascomycetes Penicillium solitum and Aspergillus niger converted alpha-farnesene to 7-hydroxyfarnesene as the major product. The radical mechanism of this bioconversion was proven by electron spin resonance (ESR) and GC-MS using the spin trapping technique. Intermediate carbon-centered radicals of alpha-farnesene were captured using two spin traps, 2-methyl-2-nitrosopropane and alpha-(4-pyridyl-1-oxide)- N- tert-butylnitrone, respectively. The evaluation of the coupling constants and hyperfine couplings of the ESR spectra showed that tertiary carbon radicals were trapped. The radical position at C7 of alpha-farnesene was derived from EI and CI mass spectra of the corresponding MNP spin adduct. The present study demonstrates that the complementary application of ESR and MS spectrometric data allows the detailed evaluation of a radical mechanism of a fungal terpene transformation reaction.

Production of Secondary Metabolites by Some Terverticillate Penicillia on Carbohydrate-Rich and Meat Substrates

Most terverticillate penicillia isolated from dry-cured meat products are toxigenic, but their ability to produce hazardous metabolites on meat-based substrates is not well known. The production of extrolites by selected terverticillate penicillia isolated from dry-cured ham has been studied on carbohydrate-rich media (malt extract agar, Czapek yeast autolysate agar, rice extract agar, and rice), meat extract triolein salt agar, and ham slices. Chloroform extracts from the selected strains grown on malt extract agar were toxic for the brine shrimp (Artemia salina) larvae and VERO cells at a concentration of 2 mg/ml, but 0.02 mg/ml produced no toxic effect. Analysis by high-pressure liquid chromatography (HPLC) coupled with photodiode array detection (DAD) or with mass spectrometry (MS) and an atmospheric pressure chemical ionization (APCI) source revealed different biologically active metabolites: cyclopiazonic acid and rugulovasine A from Penicillium commune; verrucosidin, anacine, puberuline, verrucofortine, and viridicatols from Penicillium polonicum; arisugacin and viridicatols from Penicillium echinulatum; and compactin and viridicatols from Penicillium solitum. Most of these metabolites, including the amino acid–derived compounds, were produced in the media containing high levels of carbohydrates. High concentrations of nitrogen compounds in the medium does not imply a greater production of the metabolites studied, not even those derived from the amino acids. However, molds growing on dry-cured ham are able to synthesize limited amounts of some secondary metabolites, a fact not previously reported. The combination of HPLC coupled with DAD and MS-APCI was useful for identification of closely related terverticillate Penicillium species from dry-cured ham. These techniques could be used to characterize the risk associated with the potential production of secondary metabolites in cured meats.

Solistatinol, a Novel Phenolic Compactin Analogue from Penicillium solitum.

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Mould growth on traditional greek sausages and penicillin production by Penicillium isolates

Visible moulds were isolated and identified from traditional Greek sausages from Northern Greece. Penicillium species were isolated from 90.8% of visibly mouldy sausages. Penicillium solitum, P. nalgiovense and P. commune species made up 60.6% of the total number of isolates. The most frequently occurring species was P. solitum (26.1%). P. nalgiovense and P. olsonii were found to be positive to penicillin production in an agar assay and further examination for antibiotic production in liquid culture with complex media designed for penicillin production, confirmed their ability for penicillin biosynthesis. Penicillin production by P. olsonii is reported for the first time in this study.

Solistatinol, a novel phenolic compactin analogue from Penicillium solitum

Solistatinol, a novel phenolic compactin analogue, has been isolated from Penicillium solitum using a UV-guided strategy. The structure and relative stereochemistry were determined by NMR spectroscopy and mass spectrometry. The absolute stereochemistry was determined by chemical degradation and comparison of CD data with literature data.

Control of greenhouse tomato root rot [Pythium ultimum] in hydroponic systems, using plant-growth-promoting microorganisms

Twenty-eight microorganisms showing in vitro antagonistic activity against Pythium ultimum were tested for their ability to reduce root rot [P. ultimum] on mature tomato plants grown in a greenhouse under hydroponic conditions. Of those, Penicillium brevicompactum, Penicillium solitum strain 1, Pseudomonas fluorescens subgroup G strain 2, Pseudomonas marginalis, Pseudomonas putida subgroup B strain 1, Pseudomonas syringae strain 1, and Trichoderma atroviride were shown to strongly reduce root rot severity, to improve the anchorage, and to increase the marketable yields of the plants grown in rockwool infested with P. ultimum. Experiments conducted in an organic medium containing peat, pine sawdust, and compost (v / v / v ; 60:30:10) also revealed the capability of most of these microorganisms to reduce root rot severity and to improve the anchorage of the tomato plants. However, Pseudomonas marginalis was the only microorganism that significantly improved fruit production of infected plants grown in organic medium. Moreover, Penicillium brevicompactum, Penicillium solitum strain 1, Pseudomonas fluorescens subgroup G strain 2, Pseudomonas marginalis, Pseudomonas putida subgroup B strain 1, and T. atroviride were shown to stimulate the growth of healthy tomato seedlings, suggesting that they act as PGPR (plant-growth-promoting rhizobacteria) or PGPF (plant-growth-promoting fungi). This study led to the selection of potential biocontrol agents against root rot of tomato caused by P. ultimum in hydroponic systems. This may open the way for new alternatives for the biological control of Pythium diseases in hydroponic systems that not only protect the crop but also have a beneficial effect on the plant growth and development in the absence of pathogens.