Chrysosporium Pannorum Research Articles

Survey of Keratinophilic Fungi from Feathers of Birds in Tuscany.

Simple SummaryBirds represent an effective mechanism for spreading microorganisms because they can travel great distances. The present study presents the results of an extensive survey carried out to evaluate the occurrence of keratinophilic molds on several avian species from Tuscany (central Italy). These fungal species encompass several keratin-degrading fungi, frequently responsible for skin disease in both human patients and animals. Despite the fact that molds more frequently isolated belong to dark fungi, keratinophilic species, including a dermatophyte species responsible for human and animal tinea, were cultured in large amounts, mostly from terrestrial avian species. This finding could be due to the high water content in nests of aquatic birds, which is unsuitable for the spreading of keratinophilic molds, and due to the antidermatophytic action of the larger amounts of uropygial gland secretions produced by waterfowls when compared to terrestrial birds. Our findings confirm the important role of migrating birds in the dispersal of pathogenic and saprophytic fungi to other environments, with an evolutionarily important consequence of genetic mixing of fungal population throughout the world.Although keratinophilic fungi on avian feathers have been widely described, data from European literature are quite lacking regarding Mediterranean countries. The aim of the present study was therefore to evaluate the occurrence of fungal species on feathers of different avian species in Italy. A total of 378 feather samples from both aquatic (n = 254) and terrestrial birds (n = 124), for a total of 30 bird species, were cultured for keratinophilic fungi. Fungal isolates were recognized by their macro- and micro-scopical morphology, and results were corroborated by PCR and sequencing. Keratinophilic fungi belonging to 11 different species (Scopulariopsis brevicaulis, Chrysosporium keratinophilum, Trichophyton terrestre, Microsporum gypseum, Sepedonium sp., Chrysosporium pannorum, Myriodontium sp., Chrysosporium tropicum, Chrysosporium pruinosum, Chrysosporium luteum and Aphanoascus fulvescens) were isolated from 71 animals (18.8%). The frequency of isolation of keratinophilic species from terrestrial birds was significantly higher in waterfowl. Migratory birds in Italy have been proven to carry pathogenic fungi such as dermatophytes, (A. platyrhyncos, A. crecca, E. rubecula), besides saprophytic species.

Biomass of a Psychrophilic Fungus as a Biocatalyst for Efficient Direct Esterification of Citronellol

A biomass-bound lipase from psychrophilic Chrysosporium pannorum A-1 is an efficient biocatalyst for direct esterification of β-citronellol and acetic acid in an organic solvent. The biomass is effectively produced by fungal submerged culture at 20 ℃, which results in lower energy consumption during the production of biocatalyst. Supplementation of the culture medium with calcium carbonate together with olive oil contributed to a significant increase in the active biomass of mycelium in one batch culture and increased the efficiency of the biocatalyst. Biomass-bound lipase showed high catalytic activity in a broad temperature range of 30–60 °C and stability up to 70 °C. A maximum molar conversion value of 98% was obtained at 30 °C in n-hexane using a 2:1 alcohol-to-acid molar ratio and 3% w/v of the biocatalyst within 24 h. The high equimolar concentration of the substrates (200 mM) did not have an adverse effect on mycelial biomass activity. Dry mycelium of C. pannorum is a promising biocatalyst for large-scale biosynthesis of citronellyl acetate, given its low-cost production, high activity at low temperatures, and reusability in a minimum of seven 24-h biocatalytic cycles.

Mutagenesis and Adaptation of the Psychrotrophic Fungus Chrysosporium pannorum A-1 as a Method for Improving β-pinene Bioconversion.

Mutagenesis and adaptation of the psychrotrophic fungus Chrysosporium pannorum A-1 to the toxic substrate β-pinene were used to obtain a biocatalyst with increased resistance to this terpene and improved bioconversion properties. Mutants of the parental strain were induced with UV light and N-methyl-N′-nitro-N-nitrosoguanidine. Mutants resistant to β-pinene were isolated using agar plates with a linear gradient of substrate concentrations. Active mutants were selected based on their general metabolic activity (GMA) expressed as oxygen consumption rate. Compared to the parental strain, the most active mutant showed an enhanced biotransformation ability to convert β-pinene to trans-pinocarveol (315 mg per g of dry mycelium), a 4.3-fold greater biocatalytic activity, and a higher resistance to H2O2-induced oxidative stress. Biotransformation using adapted mutants yielded twice as much trans-pinocarveol as the reaction catalyzed by non-adapted mutants. The results indicate that mutagenesis and adaptation of C. pannorum A-1 is an effective method of enhancing β-bioconversion of terpenes.

Effect of some abiotic stresses on the biotransformation of α-pinene by a psychrotrophic Chrysosporium pannorum

Biotransformation of hydrophobic terpenes is limited by their toxicity and low bioavailability to biocatalyst cells. The metabolic activity of microorganisms, which is strongly dependent on environmental parameters, may also be affected by stressful conditions. The influence of pre-incubation of the fungus Chrysosporium pannorum in different stress conditions (organic solvents, medium pH and temperatures) on its activity in the oxidative bioconversion of α-pinene to verbenone and verbenol was examined. The total bioconversion activity increased over 2-fold after 15-min pre-treatment with 1.4-dioxane as an abiotic stress factor. Also, the change of the medium pH from the optimal 5.6 to 2.0 and 10 for 1h before biotransformation enhanced product yield nearly 1.5-fold. Moreover, the use of dioxane, chloroform and ethanol and thermal stress at 50°C caused changes in the oxidation product profile (predominance of trans-pinocarveol over trans-verbenol). A maximum increase in the yield of verbenol was observed when the biocatalyst was subjected to 15-min dioxane stress before 24-h biotransformation of 1% (v/v) pinene. The significance of the data presented is that abiotic stresses may improve the bioconversion activity or create changes in the proportion of the main biotransformation products.

Bioconversion of α-pinene by a novel cold-adapted fungus Chrysosporium pannorum.

The psychrotrophic fungus Chrysosporium pannorum A-1 is reported for the first time as a novel biocatalyst for O2-promoted oxidation of α-pinene. GC–MS analysis indicated that the main products of the reaction were compounds of a high commercial value, verbenol (1) and verbenone (2). Exponentially growing cells (days 2–3) were about twice as active as cells in the late stationary phase in terms of the total concentration of products. The highest yields of 1 and 2 were obtained using three-day and two-day-old mycelia and a medium containing 1.5 and 1 % (v/v) of the substrate, respectively. The optimal time for the bioconversion of α-pinene varied from 1 to 3 days, and depended on the kind of product desired. Most of 1 was produced at a relatively high concentration of 360 mg/L after the first six hours of α-pinene bioconversion [with an average yield of 69 mg/(g dry cell L aqueous phase)]. The oxidative activity of C. pannorum was identified across a wide temperature range of 5–25 °C, 10 °C being the optimum for the production of 1 and 20 °C for the production of 2. Sequential addition of the substrate during 3 days of the biotransformation resulted in a significant increase in 1 and 2 up to 722 and 176 mg/L, respectively, and a 2-fold enhancement of product yield as compared to bioconversion with a single supply of α-pinene. The concentration of total conversion products in the culture medium reached 1.33 g/L [which corresponded product yield of 225 mg/(g dry cell L)]. This represents probably the most promising result reported to date for oxidative biotransformation of α-pinene by a wild-type microorganism.Electronic supplementary materialThe online version of this article (doi:10.1007/s10295-014-1550-0) contains supplementary material, which is available to authorized users.

If you can’t stand the heat, stay out of the city: Thermal reaction norms of chitinolytic fungi in an urban heat island

Elevated soil and air temperatures in urban heat islands have been exerting evolutionary pressure on organisms for decades in some cities. We measured thermal reaction norms (18–26 °C) for growth rate of four species of common chitinolytic fungi from an oak forest in an urban heat island and a corresponding rural area. Urban isolates of Chrysosporium pannorum and Trichoderma koningii grew faster than rural isolates at 26 °C, but grew slower than rural isolates at 18 °C. Urban isolates of Torulomyces lagena and Penicillium bilaii grew as fast or faster than rural isolates at all temperatures. These differences in thermal reaction norms between urban and rural isolates suggest that urbanization has caused both thermal specialization and counter-gradient variation in the fungal community.

Fungal bioconversion of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenol (2,4-DCP)

Ninety strains of fungi from the collection of our mycology laboratory were tested in Galzy and Slonimski (GS) synthetic liquid medium for their ability to degrade the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and its by-product, 2,4-dichlorophenol (2,4-DCP) at 100 mg l −1, each. Evolution of the amounts of each chemical in the culture media was monitored by HPLC. After 5 days of cultivation, the best results were obtained with Aspergillus penicilloides and Mortierella isabellina for 2,4-D and with Chrysosporium pannorum and Mucor genevensis for 2,4-DCP. The data collected seemed to prove, on one hand, that the strains responses varied with the taxonomic groups and the chemicals tested, and, on the other hand, that 2,4-D was less accessible to fungal degradation than 2,4-DCP. In each case, kinetics studies with the two most efficient strains revealed that there was a lag phase of 1 day before the onset of 2,4-D degradation, whereas there was none during 2,4-DCP degradation. Moreover, 2,4-DCP was detected transiently during 2,4-D degradation. Finally, M. isabellina improved its degradation potential in Tartaric Acid (TA) medium relative to GS and Malt Extract (ME) media.

Changes in Microfungal Communities in Roots of Quercus robur Stumps and their Possible Effect on Colonization by Armillaria

AbstractSixteen fungal communities were sampled by isolating from 0.5–1 cm diameter roots of living trees and stumps of common oak (Quercus robur). The density of fungi was 1.5–2 times greater in roots from stumps than from living trees. The diversity of fungi was similar in living tree roots and stumps. Some of the fungal species with increased densities in stump roots (e.g. Aspergillus kanagawaensis, Chrysosporium pannorum, Cylindrocarpon destructans, C. didymum, Hormiactis candida, Monodictys lepraria, Mycelium radicis atrovirens, Penicillium daleae, P. janczewskii, and Trichocladium opacum) usually stimulated the growth of rhizomorphs of either Armillaria ostoyae or A. gallica in oak‐wood segments in vitro. Eight of 27 isolates that were studied stimulated the rhizomorph growth in both Armillaria spp. It is presumed that the increase in density of `stimulants' may predispose oak stumps to infection by A. ostoyae and A. gallica.

Fungi of the Windmill Islands, continental Antarctica. Effect of temperature, pH and culture media on the growth of selected microfungi

Studies were conducted on microfungi isolated from soils in the Windmill Islands, continental Antarctica. Growth responses of Alternaria alternata, Chrysosporium pannorum, Nectria peziza, Thelebolus microsporus, Mycelia sterile and Phoma cf. herbarum to temperature, pH and culture media were investigated. Maximum growth occurred after 16 days, except in Nectria peziza and Thelebolus microsporus, where maximum growth occurred 12 days after inoculation. All isolates showed poor growth at 0°C. Maximum growth was obtained with temperatures ranging from 15 to 25°C. The optimum temperature for all fungi was 20°C. An acid medium favoured growth. Chrysosporium pannorum, Phoma cf. herbarum and Nectria peziza grew best at pH 3–4, whereas Mycelia sterile, Alternaria alternata and Thelebolus microsporus grew best at pH 5–6. The culture medium had little effect on growth, except for nutrient agar, which showed poor growth against all isolates with the exception of Thelebolus microsporus.

Effect of substrate and plant maturity on the incidence of infection of potato roots by pathogenic and non-pathogenic fungi

Colonization of roots of field-grown potato plants by pathogenic and non-pathogenic fungi was examined by plating surface-sterilized roots onto low nutrient medium. Plants which originated from micropropagated nodal cuttings (MP plants) or from seed-grade potato tubers (ST plants) were subjected to nine pre-plant treatments designed to test the effect of different potting substrates and different levels of natural tuber inoculum on the root mycoflora. The experiment was repeated in a second year but at a different site; in each year roots were collected once in July, August and September. Cylindrocarpon destructans, Colletotrichum coccodes, Trichoderma viride, Microdochium bolleyi and Fusarium tabacinum showed the highest mean frequencies of colonization; Verticillium tricorpus, Rhizoctonia solani, Polyscytalum pustulans and Gliocladium roseum were mainly confined to roots of ST plants or MP plants with added tuber inoculum, and Trichoderma viride, Chrysosporium pannorum, Penicillium spp., Aureobasidium pullulans and Phoma leveillei were dominant in roots of MP plants raised in peat-based potting substrates. There was evidence of suppression of the potato pathogens R. solani, P. pustulans and C. coccodes, and of several non-pathogens in roots of the same plants. Most of the fungi recovered from potato roots were also typical components of the soil microflora and occurred in both field sites. Exceptions were Fusarium oxysporum and V. dahliae, which were found in one year only. Possible associations and dissociations between different combinations of fungi occurring together on the same root system were evaluated using a correlation matrix, and spatial relationships of different groups demonstrated by principal coordinate analysis. The influence of the indigenous mycoflora on root colonization by particular potato tuber pathogens is discussed.

Pannorin, a new 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibitor produced by Chrysosporium pannorum.

Pannorin, a naphthopyrone that inhibits 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in cholesterol synthesis, was isolated from a culture broth of Chrysosporium pannorum M10539 by solvent extraction, Bio-Gel P-6 column chromatography and reverse phase HPLC (Silica ODS). Spectroscopic analyses of the compound yielded 4,8,10-trihydroxy-5-methyl-2H-naphtho[1,2-b]pyran-2-one as the proposed structure. Pannorin inhibited HMG-CoA reductase and in vitro sterol synthesis 50% at a concentration of 160 microM.

Purification and some properties of endoinulinase from Chrysosporium pannorum

One endoinulinase (2,1-β- d-fructan fructanohydrolase EC 3.2.1.7) was purified from Chrysosporium pannorum AHU 9700. The enzyme was a glycoprotein having an isoelectric point around pH 3.8. The molecular weight was 58,000 and 56,000 by SDS-polyacrylamide gel electrophoresis and gel filtration on Sephacryl S-200, respectively. The endoinulinase was most active between pH 6.0–7.0 at 50°C, and was stable at 45°C (10 min) and from pH 4.5 to 8.5 (24 h). This enzyme was active only on inulin, not on levan, sucrose, raffinose, or melezitose. The main products from inulin were inulotriose, inulotetraose, and inulopentaose.

Purification and characteristics of two exoinulinases from Chrysosporium pannorum

Two types of exoinulinase (2,1-β- d-fructan fructanohydrolase EC 3.2.1.7) were purified from Chrysosporium pannorum AHU 9700. The enzymes, F2 and F3, were glyco-proteins having isoelectric points around pH 4.6 and 4.45. The molecular weights were estimated by SDS-polyacrylamide gel electrophoresis to be 84,000 and 70,000, respectively. The enzymes were active on inulin, sucrose, raffinose, stachyose, and fructo-oligosaccharides, but not on melezitose. Levan could be hydrolyzed by exoinulinase F3, but not by exoinulinase F2. The hydrolysis products of inulin by the two enzymes were fructose and small amounts of glucose.

Inulinase from Chrysosporium pannorum

A mold, Chrysosporium pannorum AHU 9700, isolated from soil, was found to produce a very active inulin-hydrolyzing enzyme. The inulinase activity of this strain was induced by inulin, but not by sucrose, glucose, or fructose. The highest inulinase activity, 115 units/ml, was obtained under the optimum conditions. A crude enzyme preparation was active against inulin, levan, sucrose, and raffinose, but not melezitose. An inulin suspension of 10% was hydrolyzed completely at pH 5.6, 50°C, in 8 h, and inulo-oligosaccharides were detected in the initial stage of enzymatic reaction. Jerusalem artichoke extract was also rapidly hydrolyzed into fructose and small amounts of glucose. These results indicate that the C. pannorum crude enzyme contained exo- and endo-inulinases. The inulinase of C. pannorum AHU 9700 might be used industrially to prepare high fructose syrup from inulin-containing agricultural crops.

Keratinophilic fungal flora isolated from small wild mammals and rabbit-warren in France. Discussion on the fungal species found

The occurrence of dermatophytes and other keratinophilic fungi was investigated in 237 small wild mammals and 125 european rabbit. The purpose of the investigation was to determine what were the species of fungi present in the these animals. Four species of dermatophytes were isolated: Trichophyton ajelloi, Trichophyton terrestre, Trichophyton mentagrophytes, Microsporum persicolor. Trichophyton terrestre was the most frequently isolated and it occurred more frequently than its presence could be explained by the contamination from soil. Members of the genus Chrysosporium were found in many animals: Chrysosporium keratinophilum, Chrysosporium tropicum, Chrysosporium multifidum, Chrysosporium pannorum, Chrysosporium georgii, Chrysosporium merdarium, Chrysosporium anamorph of Arthroderma curreyi, Chrysosporium anamorph of Arthroderma cuniculi, Anixiopsis stercoraria, Chrysosporium parvum. Wild small mammals and european rabbits in France, not only act as carrier of keratinophilic fungi and allied dermatophytes but also provide a suitable habitat for their survival as saprophytes. The recurrence of numerous species present on the coat, isolated fort the first time in France was remarkable.

Temperature and water activity minima for growth of spoilage moulds from meat.

Five species of fungi were isolated from mould spoilage on meat other than black spot. 'White spot' colonies yielded Chrysosporium pannorum or an Acremonium sp.; 'whiskers' colonies yielded Thamnidium elegans or Mucor racemosus, and blue-green colonies yielded Penicillium corylophilum . Chrysosporium pannorum was moderately xerotolerant with a minimum growth temperature of -5 degrees C. The Acremonium sp. and P. corylophilum showed a similar level of xerotolerance but had a minimum growth temperature of -2 degrees C. Mucor racemosus was no more xerotolerant than many spoilage bacteria and did not grow below -1 degree C, but grew rapidly at 3 degrees C and above. Thamnidium elegans grew at -7 degrees C on supercooled medium and an intrinsic minimum growth temperature of -10 degrees C was indicated. However, the low xerotolerance of this species precluded growth on frozen media below -5 degrees C. It seems therefore that -5 degrees C is the practical limiting temperature for mould growth on meat, and mould spoilage usually indicates that surfaces of freezer stored meats have approached and possibly exceeded 0 degrees C.

Chryscandin, a novel peptidyl nucleoside antibiotic. I. Taxonomy, fermentation, isolation and characterization.

A novel antifungal antibiotic, chryscandin was found in the culture broth of a strain of fungi. The producing organism was subsequently identified as Chrysosporium pannorum. The antibiotic obtained as colorless needle crystals (C20H23N7O6, MW 457) is active against Candida albicans and Gram-positive bacteria, but it is ineffective against filamentous fungi and Gram-negative bacteria. Acute toxicity is very low in mice.

Structure and total synthesis of chryscandin, a new antifungal antibiotic

Abstract The structure elucidation and total synthesis of chryscandin (1), a new antifungal antibiotic produced by Chrysosporium pannorum No.4629, is reported.

Association between Chrysosporium pannorum and Mucor hiemalis in Poa flabellata litter

Mucor hiemalis was frequently isolated from the litter of Poa flabellata where it apparently grows as a secondary sugar fungus in association with Chrysosporium pannorum.

Isolation of keratinophilic fungi from sewage sludge.

Five mixed samples of sewage sludges from 4 wastewater treatment plants were examined for keratinophilic fungi. Microsporum gypseum, Trichophyton terrestre, T. ajelloi, Chrysosporium pannorum, C. keratinophilum and C. pruinosum were recovered.