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1.
Lett Appl Microbiol ; 54(5): 433-40, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22360347

RESUMEN

AIMS: To develop a novel PCR-based method able to detect potential cellulolytic filamentous fungi and to classify them exploiting the amplification of the cellobiohydrolase gene (cbh-I) and its polymorphism. METHODS AND RESULTS: A mixed approach including the combination of (i) fungal cultivation and isolation, (ii) classification of fungal isolates through the amplification of the cbh gene using a fluorescently labelled primer (f-CBH-PCR) and (iii) final fungal identification based on amplification and sequencing of the ITS1-5.8S rDNA-ITS2 region of the selected fungal strains was developed. By this approach, it was possible to screen 77 fungal strains belonging to 14 genera and 26 species. CONCLUSIONS: The f-CBH-PCR permitted the discrimination of fungal species, producing typical f-CBH profiles. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, the cbh gene was used as a preliminary classification tool able to differentiate among themselves the fungal members isolated from indoor museum items and surrounding environment. Such mixed approach consented the fast identification of all isolated fungal strains. The f-CBH-PCR method demonstrated its discrimination power, and it can be considered as a new molecular system suitable for the classification of fungal strains isolated from different environments.


Asunto(s)
Celulosa 1,4-beta-Celobiosidasa/metabolismo , Hongos/clasificación , Secuencia de Bases , Celulosa 1,4-beta-Celobiosidasa/genética , ADN de Hongos/análisis , ADN Ribosómico , Hongos/genética , Hongos/aislamiento & purificación , Datos de Secuencia Molecular , Museos , Técnicas de Tipificación Micológica/métodos , Reacción en Cadena de la Polimerasa/métodos
2.
Bioelectrochemistry ; 67(2): 191-7, 2005 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-16122687

RESUMEN

The aim of this work was voltammetric determination of 1-aminopyrene and 1-hydroxypyrene using carbon paste electrodes modified with cyclodextrin derivatives and double stranded deoxyribonucleic acid (dsDNA). The detection schemes based on a preconcentration and differential pulse voltammetric (DPV) determination at beta-cyclodextrin and gamma-cyclodextrin modified carbon paste electrode (beta-CD/CPE, gamma-CD/CPE), neutral beta-cyclodextrin polymer and carboxymethyl-beta-cyclodextrin polymer modified screen-printed electrode (beta-CDP/SPE, beta-CDPA/SPE) and dsDNA modified screen-printed electrode (DNA/SPE) are proposed for the trace determination of studied analytes within the concentration range from 2 x 10(-8) to 4 x 10(-7) mol dm(-3) and from 2 x 10(-7) to 4 x 10(-6) mol dm(-3) with the limits of quantification down to 10(-8) mol dm(-3). Depending on pH, 1-aminopyrene interacts with both surface attached CD and DNA by electrostatic bonds and supramolecular complexation while 1-hydroxypyrene associates with the CD hosts via complexation. The 1-aminopyrene interaction with dsDNA was confirmed by fluorimetric measurements in the solution phase using a competing DNA-TO-PRO-3 dye complex. In addition, the effect of temperature on this association was investigated using an electrically heated DNA-modified carbon paste electrode (DNA/CPE).


Asunto(s)
Técnicas Biosensibles/métodos , Ciclodextrinas/química , ADN/química , Electroquímica/métodos , Pirenos/análisis , Carbono/química , Electrodos , Potenciometría
3.
J Chromatogr A ; 893(1): 201-6, 2000 Sep 29.
Artículo en Inglés | MEDLINE | ID: mdl-11043601

RESUMEN

Pressurised liquid extraction (PLE) was used in the extraction of three ketones of polycyclic aromatic hydrocarbons from the sample of a soil highly contaminated with polycyclic polyaromatic compounds. The choice of solvent was the only factor that considerably influenced the extraction efficiency of PLE under the conditions recommended in Method 3545A promulgated by the United States Environmental Protection Agency. The dichloromethane-ethanol solvent mixture was found to be the most efficient solvent. PLE using this mixture provided better recoveries of all analysed ketones relative to Soxhlet extraction.


Asunto(s)
Cetonas/aislamiento & purificación , Hidrocarburos Policíclicos Aromáticos/química , Suelo/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos
5.
Fresenius J Anal Chem ; 367(4): 364-8, 2000 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-11225862

RESUMEN

Five morpholino-quinazoline derivatives have been investigated voltammetrically using a competition with the tris(o-phenanthroline) cobalt(III) redox marker for the accumulation at dsDNA modified screen-printed electrodes. An association of quinazolines with DNA was observed at the modified electrodes polarized by the negative potential of -0.4 V vs. Ag/AgCl. This was confirmed by a potentiometric stripping analysis based on the DNA guanine signal. Calibration curves for quinazolines within a concentration range of micromol/L were obtained with DP voltammetry using 5 x 10(-7) mol/L Co(phen)3(3+) marker. The quinazolines exhibit no effect on the DNA complex with the fluorescent thiazole orange derivative TO-PRO-3. The role of the accumulation potential in the association interaction with DNA is discussed.


Asunto(s)
ADN/química , Electroquímica , Electrodos , Quinazolinas/química , Calibración , Estructura Molecular
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