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PubMed1988

Enzyme-gold affinity labelling of cellulose.

The enzyme-linked colloidal gold affinity labelling technique was tested as a method to localize cellulose on thin sections of plant cell walls and slime mold spores. Commercially available cellulase from cultures of Trichoderma reesei, the main components being cellobiohydrolase I and II (CBH I, CBH II) and endoglucanase (EG), was linked to colloidal gold by using standard techniques and applied as a dilute, buffered suspension to thin sections. After brief exposure, e.g., 15-30 minutes, cellulose exposed on the surface of sections was labelled with the enzyme-gold complex. Poststaining did not appear to have a deleterious effect on the labelled sections. The specificity of labelling was demonstrated by its complete inhibition when carboxymethylcellulose was incorporated in the labelling mixture, by lack of labelling of 1,4-beta-mannans or 1,3-beta-xylans in noncellulosic walls of marine algae, by lack of labelling of 1,4-beta-glucans in chitin, by much lower labelling density when done at 4 degrees C, and by lack of labelling when sections were predigested with cellulase. Labelling with the crude commercial cellulase was compared to labelling with purified CBH I-, CBH II-, and EG-linked colloidal gold, and the labelling pattern was similar. This method was found useful on conventionally fixed material and required no special preparation other than the use of inert (Ni or Au) grids and 0.5% gelatin to reduce nonspecific binding of the gold complex. Labelling was similar in the several embedding resins tested: LR White, Lowicryl K4M, Epon 812, and Spurr's.(ABSTRACT TRUNCATED AT 250 WORDS)

Affinity Labels↗

PubMed2003

Airborne fungal spores at the Belgrad forest near the city of Istanbul (Turkey) in the year 2001 and their relation to allergic diseases.

The aim of this study was to investigate the relationship between airborne fungal spores and allergic diseases; for this reason, the airborne fungal spores that were obtained from five different locations of Belgrad Forest were isolated, determined and studied quantitatively. Totally 120 samples were examined by using the Petri Plate Gravitational Method, and fungi obtained from these samples were isolated and 600 colonies were counted. By identification of these isolations, 13 genera (Mucor, Rhizopus, Absidia, Aspergillus, Penicillium, Trichoderma, Trichothecium, Stemphylium, Cladosporium, Alternaria, Ulocladium, Aureobasidium and Fusarium), 25 species and 10 different sterile fungi were determined. The identifications of these fungi were made according to their microscopic, macromorphological features and references.

Air Microbiology↗

PubMed2005

Biodegradation study of crop residues as affected by exogenous inorganic nitrogen and fungal inoculants.

The comparative decomposition of chickpea residue, and chopped and unchopped wheat straw was investigated in pits for 120 days. Microbial biomass, humus, C/N ratio, pH, Electrical conductivity (EC), dehydrogenase, alkaline phosphatase, cellulase, xylanase, total phenol and soluble protein were determined to assess their response to the addition of inorganic nitrogen and mixed fungal inoculum of Aspergillus nidulans, Phanerochaete chrysosporium and Trichoderma viride. The evaluation of physico-chemical parameters (organic matter, organic carbon, N, C/N, pH, EC, microbial biomass) revealed that by supplementing unchopped wheat straw with 1% urea and mixed fungal inoculum, a lowest C/N ratio of 10.7, lowest biomass of 9.54 and highest humus content of 13% can be achieved within 3 months. Germination of Lepidium sativum (cress seeds) showed a germination index >60%, in this treatment. The enzyme assay for dehydrogenase indicated highest microbial activity in uninoculated treatments compared to fungal inoculated counterparts, in the second month sampling (active phase of composting). However, cellulase and xylanase activity showed an upward trend during curing phase of composting. Chickpea residue compost, though resulted in a C/N ratio of 17.3, but its germination index was less than 60%. The rapid quality tests conducted for H2S, NH3, NO3 and starch confirmed the stability and maturity of finished compost prepared from wheat straw through microbial inoculants.

Biodegradation, Environmental↗

PubMed2005

Microfungi in cultivated fields in Eskişehir provience (Turkey).

The soil microfungi flora was investigated in four locations of Eskişehir (Turkey). 56 soil samples were seasonaly collected from 14 stations in the areas of Karacahöyük, Bahçecik, OGU I, and OGU II. A total of 110 species belonging to 32 genera were encountered including Absidia, Acremonium, Alternaria, Aspergillus, Beauveria, Botryoderma, Chaetomium, Chrysosporium, Cladosporium, Eupenicillium, Eurotium, Fusarium, Geotrichum, Gliocladium, Gonytrichum, Metarrhizium, Mucor, Myrothecium, Paecilomyces, Penicillium, Phoma, Plectosphaerella, Rhizoctania, Rhizopus, Scopulariopsis, Septonema, Stachybotrys, Trichocladium, Trichoderma, Ulocladium, Verticillium, and Wardomyces. Twenty five species were more frequent (all locations) while twenty seven species were rare (only one sample). Mainly, Acremonium kiliense, Aspergillus ochraceus, A. terricola var. americanus, A. versicolor, Cladosporium cladosporioides, Fusarium oxysporum, F. solani, Gliocladium roseum, Penicillium chrysogenum, P. corylophum, P. expansum, P. griseofulvum, P. implicatum, P. restrictum, and Stachybotrys chartarum were the most common and abundant microfungi in all locations. Five species Aspergillus subsessilis, A. terreus var. africanus, Eupenicillium egyptiacum, Paecilomyces ramosus, and Penicillium novae-zeelandiae are likely to be newly recorded for Turkey. The microfungi number in Eskişehir soils was between 25,000-234,000 CFU/g (mean value at 126,375 CFU/g).

Fungi↗

PubMed2006

Isolation, identification and seasonal distribution of soilborne fungi in tea growing areas of Iyidere-Ikizdere vicinity (Rize-Turkey).

This study involves the isolation and identification of soil borne fungi from different altitudes of Iyidere-Ikizdere vicinity in Rize, Turkey. The qualitative and quantitative fungal composition of the soil of Iyidere-Ikizdere vicinity was surveyed seasonally during a year (between summer 2003 and spring 2004). The soil samples were collected from 10 different locations. Some physical and chemical properties of the soil samples were studied. Soil samples were observed to have the proper pH levels (pH 4.00-6.75) for the growth of microfungi. By using a soil dilution technique, the mycological analysis of 40 soil samples were studied at 25 cm depth. The number of microfungi ranged from 2,000-160,000 CFU g(-1). 249 different microfungi belonging to 15 genera were isolated. 45 of the isolates were Mycelia Sterilia. Penicillium, Aspergillus, Trichoderma and Fusarium were the most abundant genera (26.8%, 14.8%, 9.6% and 8.4%, respectively). Penicillium phialosporum, Penicillium trzebinskianum and Gliocladium virens were reported for the first time in Turkey.

Hydrogen-Ion Concentration↗

PubMed1988

Mycoflora of air-conditioners dust from Riyadh, Saudi Arabia.

Using the hair baiting technique, 6 genera and 14 species were collected on Sabouraud's dextrose agar from 37 dust samples from air-conditioners. The most common fungi were Chrysosporium tropicum, C. indicum, C. keratinophilum, Aspergillus flavus followed by Acremonium strictum and Scopulariopsis brevicaulis. Using the dilution-plate method, 26 genera and 52 species were collected from 37 dust samples on glucose-(23 genera and 45 species) and cellulose-(18 genera and 34 species) Czapek's agar at 28 degrees C. The most prevalent species were Aspergillus niger, A. flavus, Penicillium chrysogenum, Stachybotrys chartarum, Ulocladium atrum, Mucor racemosus and Fusarium solani and A. niger, A. flavus, Trichoderma viride, P. chrysogenum, Ulocladium atrum, Chaetomium globosum, C. spirale, Stachybotrys chartarum and Mucor racemosus on the two media, respectively.

Air Conditioning↗

PubMed1991

Mycoflora associated with five species of freshwater leeches.

Twenty-two species belonging to 11 genera of zoosporic fungi were collected from 10 freshwater sites and from five species of leeches on sesame and hemp seeds, maize grains and Pinus pollen grains as baits at 22 degrees C. Dictyuchus, Achlya, Pythium, Allomyces and Saprolegnia were the most common genera. The highest population of zoosporic fungi was estimated from Batracobdelloides tricarinata, the lowest from Barbronia assiuti. In addition fifty-three species and 1 variety representing 23 terrestrial fungal genera were identified on glucose (17 genera and 41 species) and cellulose (20 genera and 34 species + 1 variety) Czapek's Dox agar at 28 degrees C. The most common species on the two media were Aspergillus niger, A. fumigatus, A. flavus, Trichoderma harzianum, Gibberella fujikuroi and Penicillium chrysogenum. Some fungi were common only on cellulose agar medium such as Botryotrichum atrogriseum, Chaetomium bostrychodes, Dactylella alaskana and Drechslera halodes. Batracobdelloides tricarinata was the richest with terrestrial fungi whereas Salifa delicata was the poorest. Five leech species, namely Alboglossiphonia polypompholyx, Batracobdelloides tricarinata, Helobdella conifera, Barbronia assiuti and Salifa delicata were used during this investigation.

Animals↗

PubMed1996

Diversity of microfungi in the sandy soil of Ipanema Beach, Rio de Janeiro, Brazil.

The mycobiota of the sandy soil of Ipanema Beach, Rio de Janeiro, Brazil, was investigated in 144 sand samples collected at four different sites along the sea coast, divided into three subsites, for a period of one year. A total of 4285 yeast colonies and of 6956 of colonies filamentous fungi were isolated using conventional media and techniques. Representatives of the filamentous fungi corresponding to a total of 1334 colonies were identified and assigned to 34 genera and 170 species. The genera of highest incidence and their respective numbers of species were as follows: Aspergillus, 30.4%, 32 spp.; Penicillium, 16.2%, 35 spp.; Fusarium, 12,6%, 33 spp.; Trichoderma, 6.4%, 7 spp.; Paecilomyces, 3.7%, 10 spp.; Cladosporium, 3.1%, 8 spp. and Acremonium, 1.0%, 8 spp. Several other genera and species were detected at quite low occurrences. Non-sporulating fungi (18.3%) and Coelomycetes (Sphaeropsidales) (1.9%) were also detected. Most of the genera detected belonged to the Deuteromycotina, with fewer proportions belonging to the Ascomycotina and Zygomycotina. Moniliaceae was represented by the largest number of species and Dematiaceae was represented by the largest number of genera. In terms of seasonal distribution, the largest number of filamentous fungi was isolated during the winter and the smallest number the Summer.

Fungi↗

PubMed1975

Amino acid analogs. III: New syntheses of monomethyl- and monophenylglutamic acids.

Glutamic acid analogs containing 3- and 4-methyl and 2-, 3-, and 4-phenyl substituents were prepared. The 3- and 4-methyl- and 3- and 4-phenylglutamic acids did not inhibit Plasmodium berghei and were nontoxic to the host (mice) at 640 mg/kg. The five analogs in addition to 2-methlglutamic acid were inactive against Lactobacillus casei at 1000 mug/ml in a defined medium: against Escherichia coli, only 2-methylglutamic acid caused 27% inhibition at 10,000 mug/ml. All six analogs failed to inhibit Aspergillus niger, Aspergillus oryzae, Trichoderma viride, and Myrothecium verrucaria in a defined medium below 10,000 mug/ml.

Animals↗

PubMed1978

Amino acid analogs IV:4-fluoroisoleucine.

4-Fluoroisoleucine was produced by ammonolysis of 2-bromo-4-fluoro-3-methylpentanoic acid, which resulted from the bromofluorination of 4-methyl-2-pentenoic acid. It did not inhibit Plasmodium berghei in mice at 640 mg/kg and was not toxic to the animals. The fluoroamino acid inhibited Aspergillus niger, Trichoderma viride, Myrothecium verrucaria, Trichophyton mentagrophytes, and Mucor mucedo in Czapek solution agar at a concentration between 10(4) and 10(3) microgram/ml. Growth of Escherichia coli was inhibited 25% at 900 microgram/ml in a defined medium.

Animals↗

PubMed1980

Antifungal properties of n-alkanols, alpha, omega-n-alkanediols, and omega-chloro-alpha-alkanols.

Fourteen n-alkanols (C1-C12, C14, and C16), 13 alpha,omega-n-alkanediols (C2-C12, C14, and C16), and 13 omega-chloro-alpha-alkanols (C2-C12, C14, and C16) were tested against Aspergillus niger, Trichoderma viride, and Myrothecium verucaria in Sabouraud dextrose agar at pH 4.0 and 5.6. Toxicity to Candida blbicans, Trichophyton mentagrophytes, and Mucor mucedo was determined in the same medium at pH 5.6 and 7.0 in the absence and presence of 10% beef serum. The fungitoxicity of these alcohols was influenced by chain length and insignificantly by the pH of the medium and the presence of beef serum. The C10-member of each series was most active; the order of activity of the three groups was chloroalkanols greater than alkanols greater than alkanediols. Compared to the fatty acids, the order of fungitoxicity on a weight basis was 2-alkynoic acids greater than 2-alkenoic acids greater than omega-chloralkanols greater than alkanoic acids greater than 2-bromoalkanoic acids greater than 2-fluoroalkanoic acids greater than n-alkoxyacetic acids greater than n-alkanols greater than alpha,omega-n-alkanediols.

Alcohols↗

PubMed1984

Antifungal properties of 2-n-alkyn-1-ols.

Fourteen 2-n-alkynols (C3-C14, C16, and C18) were tested against Aspergillus oryzae, Aspergillus niger, Trichoderma viride, and Myrothecium verrucaria in Sabouraud dextrose agar at pH 5.6 and 7.0. Toxicity to Candida albicans, Candida tropicalis, Trichophyton mentagrophytes, and Mucor mucedo was determined in the same medium at pH 5.6 and 7.0 in the absence and presence of 10% beef serum. Fungitoxicity was strongly influenced by chain length, slightly by the pH of the medium, and significantly by the presence of beef serum. 2-n-Undecyn-1-ol was the most active member of the series, and there was marked synergism between it and ketoconazole.

Alkynes↗

PubMed1985

Antifungal properties of 3-n-alkyn-1-ols and synergism with 2-n-alkyn-1-ols and ketoconazole.

Twelve 3-n-alkyn-1-ols (C4-C12, C14, C16, and C18) were tested against Aspergillus oryzae, Aspergillus niger, Trichoderma viride, and Myrothecium verrucaria in Sabouraud dextrose agar at pH 5.6 and 7.0. Toxicity to Candida albicans, Candida tropicalis, Trichophyton mentagrophytes, and Mucor mucedo was determined in the same medium at pH 5.6 and 7.0 in the absence and presence of 10% beef serum. Fungitoxicity was strongly influenced by chain length, slightly by pH of the medium, and significantly but not strongly by the presence of beef serum. 3-n-Decyn-1-ol, 3-n-undecyn-1-ol, and 3-n-dodecyn-1-ol were the most active members of the series. Synergism toward C. albicans and C. tropicalis was observed between 3-n-undecyn-1-ol and ketoconazole, and a mixture of 3-n-undecyn-1-ol, 2-n-undecyn-1-ol, and ketoconazole in Sabouraud dextrose agar at pH 7.0 in the presence of 10% human serum.

Alcohols↗

PubMed1989

Synergistic antifungal action of 8-quinolinol and its bischelate with copper(II) and with mixed ligand chelates composed of copper(II), 8-quinolinol, and aromatic hydroxy acids.

Antifungal studies were made of mixtures of minimal inhibitory concentrations (MICs) of 8-quinolinol and its bischelates with copper(II), zinc(II), and manganese(II) and with mixed ligand chelates composed of 8-quinolinol, copper(II) and a second ligand including salicylic acid, 3-hydroxy-2-naphthoic acid, 3,5-diiodosalicylic acid, and 4-bromo-3-hydroxy-2-naphthoic acid. Mixtures of the MICs of the bischelates of 8-quinolinol with copper(II) and zinc(II) and copper(II) and manganese(II), as well as 7-iodo-8-quinolinol and its bischelate with copper(II), and 8-quinolinol and 5-iodo-8-quinolinol were also studied against six fungi: Aspergillus niger, Aspergillus oryzae, Trichoderma viride, Myrothecium verrucaria, Mucor cirinelloides, and Trichophyton mentagrophytes. With the exceptions of the mixtures of 8-quinolinol and (8-quinolinolato)(3,5-diiodosalicylato)copper(II) and (8-quinolinolato)(4-bromo-3-hydroxy-2-naphthoato)copper(II) against M. cirinelloides, all of the test organisms were inhibited by 40% or less of each mixture containing 8-quinolinol. Bischelates of 8-quinolinol with copper(II) and zinc(II) and copper(II) and manganese(II) inhibited five fungi at 50% of the mixtures of the MICs. Mucor cirinelloides was not inhibited by bis(8-quinolinolato)copper(II), bis(8-quinolinolato)zinc(II), or by bis(7-iodo-8-quinolinolato)copper(II).(ABSTRACT TRUNCATED AT 250 WORDS)

Antifungal Agents↗

PubMed2006

Characterization of filamentous fungi isolated from Moroccan olive and olive cake: toxinogenic potential of Aspergillus strains.

During the 2003 and 2004 olive oil production campaigns in Morocco, 136 samples from spoiled olive and olive cake were analyzed and 285 strains were isolated in pure culture. Strains included 167 mesophilic strains belonging to ten genera: Penicillium, Aspergillus, Geotrichum, Mucor, Rhizopus, Trichoderma, Alternaria, Acremonium, Humicola, Ulocladium as well as 118 thermophilic strains isolated in 2003 and 2004, mainly belonging to six species: Aspergillus fumigatus, Paecilomyces variotii, Mucor pusillus, Thermomyces lanuginosus, Humicola grisea, and Thermoascus aurantiacus. Penicillium and Aspergillus, respectively, 32.3 and 26.9% of total isolates represented the majority of mesophilic fungi isolated. When considering total strains (including thermotolerant strains) Aspergillus were the predominant strains isolated; follow-up studies on mycotoxins therefore focused primarily on aflatoxins (AFs) and ochratoxin A (OTA) from the latter strains. All isolated Aspergillus flavus strains (9) and Aspergillus niger strains (36) were studied in order to evaluate their capacity to produce AFs and OTA, respectively, when grown on starch-based culture media. Seven of the nine tested A. flavus strains isolated from olive and olive cake produced AF B1 at concentrations between 48 and 95 microg/kg of dry rice weight. As for the A. niger strains, 27 of the 36 strains produced OTA.

Aflatoxins↗

PubMed2006

Detection of new amino acid sequences of alamethicins F30 by nonaqueous capillary electrophoresis-mass spectrometry.

The microheterogeneous alamethicin F30 (ALM F30) isolated from the fermentation of Trichoderma viride strain NRRL 3199 was analyzed by nonaqueous capillary electrophoresis coupled to electrospray ion-trap mass spectrometry (ESI-IT-MS) and electrospray time-of-flight mass spectrometry (ESI-TOF-MS). Tandem ESI-IT-MS was used for elucidation of the amino acid sequence based on the fragmentation pattern of selected parent ions. The MS/MS spectra using the [M + 3H](3+) or [M + 2H](2+) ions as precursor ions displayed the respective b- and the y-type fragments resulting from cleavage of the particularly labile Aib-Pro bond. The MS(3) of these fragments generated the b acylium ion series, as well as internal fragment ion series. Eleven amino acid sequences were identified, characterized by the exchange of Ala to Aib in position 6, Gln to Glu in positions 7 or 19 as well as the loss of the C-terminal amino alcohol. In addition, two truncated pyroglutamyl peptaibols were found. Overall, seven new sequences are reported compared to earlier LC-MS studies. The composition of the components was confirmed by on-line ESI-TOF-MS detection. Mass accuracy well below 5 ppm was observed. Quantification of the individual components was achieved by a combination of UV and TOF-MS detection.

Alamethicin↗

PubMed2006

Tandem electrospray mass spectrometric studies of proton and sodium ion adducts of neutral peptides with modified N- and C-termini: synthetic model peptides and microheterogeneous peptaibol antibiotics.

The fragmentations of [M+H]+ and [M+Na]+ adducts of neutral peptides with blocked N- and C-termini have been investigated using electrospray ion trap mass spectrometry. The N-termini of these synthetically designed peptides are blocked with a tertiarybutyloxycarbonyl (Boc) group, and the C-termini are esterified. These peptides do not possess side chains that are capable of complexation and hence the backbone amide units are the sole sites of protonation and metallation. The cleavage patterns of the protonated peptides are strikingly different from those of sodium ion adducts. While the loss of the N-terminal blocking group occurs quite readily in the case of MS/MS of [M+Na]+, the cleavage of the C-terminal methoxy group seems to be a facile process in the case of MS/MS of [M+H]+ * Fragmentation of the protonated adducts yields only bn ions, while yn and a(n) ions are predominantly formed from the fragmentation of sodium ion adducts. The a(n) ions arising from the fragmentation of [M+Na](+) lack the N-terminal Boc group (and are here termed a(n)* ions). MS/MS of [M+Na]+ species also yields b(n) ions of substantially lower intensities that lack the N-terminal Boc group (b(n)*). A similar distinction between the fragmentation patterns of proton and sodium ion adducts is observed in the case of peptides possessing an N-terminal acetyl group. An example of the fragmentation of the H+ and Na+ adducts of a naturally occurring peptaibol from a Trichoderma species confirms that fragmentation of these two ionized species yields complementary information, useful in sequencing natural peptides. Inspection of the isotopic pattern of b(n) ions derived from [M+H]+ adducts of peptaibols provided insights into the sequences of microheterogeneous samples. This study reveals that the combined use of protonated and sodium ion adducts should prove useful in de novo sequencing of peptides, particularly of naturally occurring neutral peptides with modified N- and C-termini, for example, peptaibols.

Amino Acid Sequence↗

PubMed1999

Cloning and characterization of a thermostable cellobiose dehydrogenase from Sporotrichum thermophile.

Cellobiose dehydrogenase (CDH) is an extracellular hemoflavoenzyme produced by several wood-degrading fungi. CDH contains one heme b and one FAD per molecule and oxidizes cellobiose to cellobionolactone in the presence of cytochrome c. In this report, a thermostable CDH from the thermophilic ascomycete Sporotrichum thermophile has been purified, cloned, and characterized. The temperature optimum for this CDH reaction was 60 degrees C, and the activation energy for the reaction was 26.3 kJ/mol. The Km and kcat were temperature-dependent and increased as reaction temperature increased. These kinetic properties prove that this CDH is truly thermophilic. A 2.8-kb cDNA was isolated by screening an expression library of S. thermophile with a polyclonal antisera raised against Phanerochaete chrysosporium CDH. The cDNA encoded an 807-amino-acid protein with a predicted mass of 86,332 Da. S. thermophile CDH is organized into three domains, an N-terminal flavin domain, a middle heme domain, and a C-terminal cellulose-binding domain, which shows sequence similarity with the cellulose-binding domains of endoglucanases and cellobiohydrolases from Trichoderma reesei. Comparison with the CDH sequences of P. chrysosporium and Trametes versicolor identified Met 95 and His 143 as potential heme coordinations. EFIG, LGGPM, and VNSTH motifs in the heme domain and the XRXPXTDXPSXDGXRY motif in the flavin domain were identified as CDH-specific motifs. With regard to the amino acid composition, S. thermophile CDH has more disulfide linkages and acidic and basic amino acids compared to CDHs from P. chrysosporium and T. versicolor.

Amino Acid Sequence↗