Selection and molecular identification of fungal isolates that produce xylanolytic enzymes.

Xylan is a heteropolysaccharide and its complete hydrolysis involves a complex set of xylanolytic enzymes. Fungal xylanases have been widely used in the holocellulose industry to obtain by-products or for its elimination. The aim of this study was to select and identify filamentous fungi from different ecosystems that produce extracellular xylanases showing biotechnological potential. One hundred three fungal isolates were obtained from orchard, horticultural, and forest ecosystems. The ability of fungi to degrade xylan was measured by quantifying their xylanolytic indices after growth on solid culture media and their extracellular xylanolytic and cellulolytic activities after submerged fermentation. All fungal isolates grew on solid medium supplemented with xylan as the sole carbon source, but only 44% of isolates showed xylanolytic indices greater than 1.0. In submerged fermentation, 39% of the fungi tested showed no cellulolytic activity. Filamentous fungi were chosen from correspondence analysis and were identified by molecular tools using internal transcribed spacers. One of the 9 isolates selected belonged to the Phoma genus and the remaining were from the Fusarium genus. Fusarium solani (isolate 59) showed the highest xylanolytic index (0.964 ± 0.042), rapid growth on solid medium (1.233 ± 0.050 cm/day), significant xylanolytic activity (3.823 ± 0.210 U/mg), and a total deficiency of cellulolytic activity compared to other fungal isolates. In the zymogram, a clear zone was observed, indicating that F. solani possesses at least 1 xylanase. Fusarium solani was selected for its ability to produce extracellular xylanases with biotechnological potential.

MRNA and miRNA expression patterns associated to pathways linked to metal mixture health effects.

Metals are a threat to human health by increasing disease risk. Experimental data have linked altered miRNA expression with exposure to some metals. MiRNAs comprise a large family of non-coding single-stranded molecules that primarily function to negatively regulate gene expression post-transcriptionally. Although several human populations are exposed to low concentrations of As, Cd and Pb as a mixture, most toxicology research focuses on the individual effects that these metals exert. Thus, this study aims to evaluate global miRNA and mRNA expression changes induced by a metal mixture containing NaAsO2, CdCl2, Pb(C2H3O2)2·3H2O and to predict possible metal-associated disease development under these conditions. Our results show that this metal mixture results in a miRNA expression profile that may be responsible for the mRNA expression changes observed under experimental conditions in which coding proteins are involved in cellular processes, including cell death, growth and proliferation related to the metal-associated inflammatory response and cancer.

Isolation and molecular characterization of a novel strain of Bacillus with antifungal activity from the sorghum rhizosphere.

We looked for bacterial strains with antifungal activity in the sorghum rhizosphere. A prescreening procedure to search for hemolytic activity among the isolated strains allowed us to detect good fungitoxic activity in a bacterial isolate that we named UM96. This bacterial isolate showed strong growth inhibition in bioassays against the pathogens Diaporthe phaseolorum, Colletotrichum acutatum, Rhizoctonia solani, and Fusarium oxysporum. The supernatant of isolate UM96 also showed strong hemolytic activity, which was not observed in the protease-treated supernatant. However, the supernatant that was treated with protease had similar antagonistic effects to those exhibited by the supernatant that was not treated with this enzyme. These results suggest that a bacteriocin-like compound is responsible for the hemolytic activity; whereas, as far as antifungal effect is concerned, an antibiotic of nonribosomal origin, such as a lipopeptide, might be acting. Further molecular characterization by partial 16S rDNA sequencing placed isolate UM96 in a cluster with Bacillus amyloliquefaciens; however, the highest identity match found in databases of Bacillus species was 91% identity. This suggests that Bacillus sp UM96 might be a novel species.

Hexavalent-chromium reduction by a chromate-resistant Bacillus sp. strain.

Bacillus strain QC1-2, isolated from a chromium-polluted zone, was selected by its high ability to both tolerate and reduce hexavalent chromium [Cr(VI)] to less-toxic trivalent chromium [Cr(III)]. Cell suspensions of strain QC1-2 rapidly reduced Cr(VI), in both aerobic and anaerobic conditions, to Cr(III) which remained in the supernatant. Cr(VI) reduction was dependent on the addition of glucose but sulfate, an inhibitor of chromate transport, had no effect. Studies with permeabilized cells and cell extracts showed that the Cr(VI) reductase of strain QC1-2 is a soluble NADH-dependent enzyme.

Inhibition of the yeast-mycelial transition and the phorogenesis of Mucorales by diamino butanone.

Diamino butanone (DAB), a competitive inhibitor of ornithine decarboxylase (ODC) a key enzyme in polyamine biosynthesis, inhibited the yeast to hyphae transition in Mucor rouxii, induced by transfer from anaerobiosis to aerobiosis, but not the opposite phenomenon. Addition of DAB to anaerobic yeast cells brought about a decrease in ODC and polyamine levels. In these conditions, the aerobic shift produced only a weak increase in ODC activity and no change in polyamine levels. DAB also blocked phorogenesis in M. rouxii and in Phycomyces blakesleeanus. At the effective concentrations DAB did not affect cell growth of either fungus. It is suggested that low, constant levels of ODC and polyamines are necessary for cell growth, and that high transient levels are required during the differentiative steps. DAB, at the concentrations used, affects this last process, but does not interfere with the maintenance level of polyamines.