Physico-chemical and microbial perturbations of Andalusian pine forest soils following a wildfire.

Wildfires are a recurrent disturbance in Mediterranean forests, triggered by high fuel load, high environmental temperature and low humidity. Although, human intervention is behind the initiation of most fire episodes, the situation is likely to worsen in the future due to the effects of climate change in the Mediterranean "hot-spot". Here we study chemical, physical and microbial characteristics of burnt soils from two well differentiated sites at Sierra de Cazorla, Segura and Las Villas Natural Park, Andalusia, (Spain) affected and unaffected by a wildfire, and followed their evolution for three years. The soils affected by a severe surface burn showed a significant increase in organic matter after 3years from the fire. Viable bacteria and fungi also increased, especially 2-3years post-burning. Substrate induced respiration (SIR) also increased significantly in burnt soil from site 1 (rendzina on carbonate) while a significant decrease was observed in the burnt soils sampled from site 2 (calcic luvisols) in samples taken one month after the wildfire. A recovery in both SIR and organic matter was observed after 2 and 3years. Of seven soil enzymes studied, only phosphatase activity was significantly higher in most burnt soils over the three years. Analysis of bacterial community diversity using clone libraries showed a recovery in the number of phyla in burnt soils after 2 and 3years in both sites, with an increase in Proteobacteria and Firmicutes and a decrease in Acidobacteria phyla. For Bacteroidetes, the percentages were lower in most burnt samples. This study reveals that if wildfire increases the organic matter availability, then the microbial community responds with increased activity and biomass production. Although fire exerts an initial impact on the soil bacterial community, its structure and functional profile soon recovers (after 2-3years) contributing to soil recovery.

Laccase SilA from Streptomyces ipomoeae CECT 3341, a key enzyme for the degradation of lignin from agricultural residues?

The role of laccase SilA produced by Streptomyces ipomoeae CECT 3341 in lignocellulose degradation was investigated. A comparison of the properties and activities of a laccase-negative mutant strain (SilA-) with that of the wild-type was studied in terms of their ability to degrade lignin from grass lignocellulose. The yields of solubilized lignin (acid precipitable polymeric lignin, APPL) obtained from wheat straw by both strains in Solid State Fermentation (SSF) conditions demonstrated the importance of SilA laccase in lignin degradation with the wild-type showing 5-fold more APPL produced compared with the mutant strain (SilA-). Analytical pyrolysis and FT-IR (Fourier Transform Infrared Spectroscopy) confirmed that the APPL obtained from the substrate fermented by wild-type strain was dominated by lignin derived methoxyphenols whereas those from SilA- and control APPLs were composed mainly of polysaccharides. This is the first report highlighting the role of this laccase in lignin degradation.

The degradation of two fluoroquinolone based antimicrobials by SilA, an alkaline laccase from Streptomyces ipomoeae.

The presence of fluoroquinolone based antimicrobials in natural waters represents a significant emerging environmental problem. In this study the suitability of a novel alkaline bacterial laccase, SilA, from Streptomyces ipomoeae to degrade two key antimicrobials, Ciprofloxacin and Norfloxacin under alkaline conditions in the presence of natural mediators was assessed. Results showed that only the selected SilA-acetosyringone system was able to degrade more than 90% of both fluoroquinolones. HPLC analysis of the degradation products obtained after enzyme treatment confirmed the disappearance of the antimicrobials and the mediator after 24 h. The time course of the degradation showed that during the first 4 h a 75% of degradation of fluoroquinolones was detected while the mediator remained stable. A concomitant appearance of new chromatographic peaks derived from the fluoroquinolones and/or the mediator was detected. Moreover, toxicity assays demonstrated that the SilA-acetosyringone system was able to reduce the toxicity of Ciprofloxacin and Norfloxacin by 90 and 70%, respectively. In conclusion, these findings support the suitability of a low cost and environmentally friendly strategy based on the SilA-acetosyringone system for a primary treatment of contaminated alkaline wastewaters with this type of emerging pollutants.

Contributions to a better comprehension of redox-mediated decolouration and detoxification of azo dyes by a laccase produced by Streptomyces cyaneus CECT 3335.

The ability of a laccase (EC 1.10.3.2) produced by Streptomyces cyaneus CECT 3335 to decolourise and detoxify azo dyes was assessed. Results showed that a colour loss of 90% was achieved only in the presence of acetosyringone (0.1mM) acting as a redox mediator for the laccase. Toxicological analysis of the decolourised dyes revealed that there was no direct correlation between decolouration and detoxification; in fact, in the case of the dyes Methyl Orange and Orange II, a significant increase in toxicity was produced after the treatment. In contrast, a significant decrease in toxicity was observed after the decolouration of New Coccine and Chromotrope 2R. Finally, HPLC analysis of the dyes after treatment revealed the complete disappearance of both dyes and mediator and a concomitant appearance of new chromatographic peaks which could be responsible of the residual toxicity detected in some cases.

Detoxification of azo dyes by a novel pH-versatile, salt-resistant laccase from Streptomyces ipomoea.

A newly identified extracellular laccase produced by Streptomyces ipomoea CECT 3341 (SilA) was cloned and overexpressed, and its physicochemical characteristics assessed together with its capability to decolorize and detoxify an azotype dye. Molecular analysis of the deduced sequence revealed that SilA contains a TAT-type signal peptide at the N-terminus and only two cupredoxine domains; this is consistent with reports describing two other Streptomyces laccases but contrasts with most laccases, which contain three cupredoxine domains. The heterologous expression and purification of SilA revealed that the homodimer is the only active form of the enzyme. Its stability at high pH and temperature, together with its resistance to high concentrations of NaCl and to typical laccase inhibitors such as sodium azide confirmed the unique properties of this novel laccase. The range of substrates that SilA is able to oxidize was found to be pH-dependent; at alkaline pH, SilA oxidized a wide range of phenolic compounds, including the syringyl and guayacil moieties derived from lignin. The oxidative potential of this enzyme to use phenolic compounds as natural redox mediators was shown through the coordinated action of SilA and acetosyringone (as mediator), which resulted in the complete detoxification of the azo-type dye Orange II.

Detoxification of azo dyes by a novel pH-versatile, salt-resistant laccase from Streptomyces ipomoea

A newly identified extracellular laccase produced by Streptomyces ipomoea CECT 3341 (SilA) was cloned and overexpressed, and its physicochemical characteristics assessed together with its capability to decolorize and detoxify an azotype dye. Molecular analysis of the deduced sequence revealed that SilA contains a TAT-type signal peptide at the N-terminus and only two cupredoxine domains; this is consistent with reports describing two other Streptomyces laccases but contrasts with most laccases, which contain three cupredoxine domains. The heterologous expression and purification of SilA revealed that the homodimer is the only active form of the enzyme. Its stability at high pH and temperature, together with its resistance to high concentrations of NaCl and to typical laccase inhibitors such as sodium azide confirmed the unique properties of this novel laccase. The range of substrates that SilA is able to oxidize was found to be pH-dependent; at alkaline pH, SilA oxidized a wide range of phenolic compounds, including the syringyl and guayacil moieties derived from lignin. The oxidative potential of this enzyme to use phenolic compounds as natural redox mediators was shown through the coordinated action of SilA and acetosyringone (as mediator), which resulted in the complete detoxification of the azo-type dye Orange II (AU)

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Soil health -- a new challenge for microbiologists and chemists.

Soil health refers to the biological, chemical, and physical features of soil that are essential to long-term, sustainable agricultural productivity with minimal environmental impact. Thus, soil health provides an overall picture of soil functionality. Although it cannot be measured directly, soil health can be inferred by measuring specific soil properties (e.g. organic matter content) and by observing soil status (e.g. fertility). There is also increased interest in studying soil microorganisms in their particular environments, as microbial diversity is intimately related to soil structure and function. One of the key objectives in determining soil health is to acquire indicators that can be used to evaluate the soil's current status and hence to develop sustainable agricultural systems. In this regard, significant progress has been made over the last few years in the development of specific biomarkers and macromolecular probes, enabling rapid and reliable measurements of soil microbial communities. In addition, modern molecular biological techniques, such as fluorescence in situ hybridization (FISH), reverse transcriptase polymerase chain reaction (RT-PCR), denaturing gradient gel electrophoresis (DGGE), and terminal restriction fragment length polymorphism (T-RFLP), have facilitated the analysis of microbial biodiversity and activity, whereas the application of modern analytical techniques, such as nuclear magnetic resonance (NMR) and pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS), have provided data on soil chemistry. The combination of these two approaches offers promise in determining soil health status.

Soil health - a new challenge for microbiologists and chemists / La salud del suelo - un nuevo reto para microbiólogos y químicos

Soil health refers to the biological, chemical, and physical features of soil that are essential to long-term, sustainable agricultural productivity with minimal environmental impact. Thus, soil health provides an overall picture of soil functionality. Although it cannot be measured directly, soil health can be inferred by measuring specific soil properties (e.g. organic matter content) and by observing soil status (e.g. fertility). There is also increased interest in studying soil microorganisms in their particular environments, as microbial diversity is intimately related to soil structure and function. One of the key objectives in determining soil health is to acquire indicators that can be used to evaluate the soil’s current status and hence to develop sustainable agricultural systems. In this regard, significant progress has been made over the last few years in the development of specific biomarkers and macromolecular probes, enabling rapid and reliable measurements of soil microbial communities. In addition, modern molecular biological techniques, such as fluorescence in situ hybridization (FISH), reverse transcriptase polymerase chain reaction (RT-PCR), denaturing gradient gel electrophoresis (DGGE), and terminal restriction fragment length polymorphism (T-RFLP), have facilitated the analysis of microbial biodiversity and activity, whereas the application of modern analytical techniques, such as nuclear magnetic resonance (NMR) and pyrolysis gas chromatography-mass spectrometry (Py-GC-MS), have provided data on soil chemistry. The combination of these two approaches offers promise in determinings oil health status (AU)

El concepto de salud del suelo se relaciona con las características biológicas, químicas y físicas que son esenciales para una productividad agrícola sostenible a largo plazo con un mínimo impacto ambiental. La salud del suelo es el más fiel reflejo de su funcionalidad. Aunque no puede medirse directamente, se puede inferir a partir de la determinación de propiedades específicas del mismo suelo (p.e. el contenido en materia orgánica) y por la observación de su estado (p.e. la fertilidad). El interés por el estudio de los microorganismos del suelo en su propio medio está aumentando, puesto que la diversidad microbiana está estrechamente relacionada con la estructura y función del suelo. Un objetivo clave para determinar la salud de un suelo es la disponibilidad de indicadores factibles de ser utilizados en la evaluación de su estado y, a partir de aquí, implementar sistemas de agricultura sostenible. El desarrollo de biomarcadores específicos y sondas macromoleculares ha evolucionado considerablemente en los últimos años, lo cual ha permitido obtener medidas fiables y rápidas de las comunidades microbianas del suelo. Por otro lado, las modernas técnicas biomoleculares más recientes (p.e. la hibridación in situ mediante fluorescencia [FISH], la reacción en cadena de la polimerasa mediante transcriptasa inversa [RT-PCR], la electroforesis en gel con gradiente desnaturalizante [DGGE]y el análisis del polimorfismo en la longitud de los fragmentos de restricción terminales [T-RFLP]) permiten analizar la biodiversidad y actividad microbianas, mientras que la aplicación de técnicas analíticas modernas (p.e. resonancia magnética nuclear [NMR], pirolisis-cromatografía de gases-espectrometría de masas [Py-GC-MS]) proporciona datos sobre la química del suelo. La combinación de estas dos aproximaciones metodológicas ofrece buenas perspectivas en la determinación del estado de salud del suelo (AU)

Kraft pulp biobleaching and mediated oxidation of a nonphenolic substrate by laccase from Streptomyces cyaneus CECT 3335.

A new laccase (EC 1.10.3.2) produced by Streptomyces cyaneus CECT 3335 in liquid media containing soya flour (20 g per liter) was purified to homogeneity. The physicochemical, catalytic, and spectral characteristics of this enzyme, as well as its suitability for biobleaching of eucalyptus kraft pulps, were assessed. The purified laccase had a molecular mass of 75 kDa and an isoelectric point of 5.6, and its optimal pH and temperature were 4.5 and 70 degrees C, respectively. The activity was strongly enhanced in the presence of Cu(2+), Mn(2+), and Mg(2+) and was completely inhibited by EDTA and sodium azide. The purified laccase exhibited high levels of activity against 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) and 2,6-dimethoxyphenol and no activity against tyrosine. The UV-visible spectrum of the purified laccase was the typical spectrum of the blue laccases, with an absorption peak at 600 nm and a shoulder around 330 to 340 nm. The ability of the purified laccase to oxidize a nonphenolic compound, such as veratryl alcohol, in the presence of ABTS opens up new possibilities for the use of bacterial laccases in the pulp and paper industry. We demonstrated that application of the laccase from S. cyaneus in the presence of ABTS to biobleaching of eucalyptus kraft pulps resulted in a significant decrease in the kappa number (2.3 U) and an important increase in the brightness (2.2%, as determined by the International Standard Organization test) of pulps, showing the suitability of laccases produced by streptomycetes for industrial purposes.