A series of polyvinyl alcohol (PVA), PVA/chitosan (CS) and PVA/cyanobacterial extracellular polymeric substances (EPS) blended nanofibrous membranes were produced by electrospinning using a microfiltration poly(vinylidene fluoride) (PVDF) basal membrane, for potential applications in water filtration. Nanofibres were obtained from solutions of 20% (w/w) PVA with 1% (w/w) CS or EPS, using a weight ratio of 60/40. Blended nanofibres have shown a smooth morphology, no beads formation and diameters between 50 and 130 nm. Thermo-mechanical analysis demonstrated that there were inter and/or intramolecular hydrogen bonds between the molecules of PVA/CS and PVA/EPS in the blends. The electrospun blended PVA/EPS membrane showed better tensile mechanical properties when compared with PVA and PVA/CS, and resisted more against disintegration in the temperature range between 10 and 50 °C. Finally, the blended membranes have shown an increase in chromium binding capacity of 5%. This is the first successful report of a blended membrane of electrospinned cyanobacterial polysaccharide with PVA.

Cyanobacterial extracellular polymeric substances (EPS) are heteropolysaccharides that possess characteristics suitable for industrial applications, notably a high number of different monomers, strong anionic nature and high hydrophobicity. However, systematic studies that unveil the conditions influencing EPS synthesis and/or its characteristics are mandatory. In this work, Cyanothece sp. CCY 0110 was used as model organism. Our results revealed that this strain is among the most efficient EPS producers, and that the amount of RPS (released polysaccharides) is mainly related to the number of cells, rather than to the amount produced by each cell. Light was the key parameter, with high light intensity enhancing significantly RPS production (reaching 1.8 g L(-1)), especially in the presence of combined nitrogen. The data showed that RPS are composed by nine different monosaccharides (including two uronic acids), the presence of sulfate groups and peptides, and that the polymer is remarkably thermostable and amorphous in nature.

Here we report on the functional characterization of the hypothetical protein Slr1270, a TolC homologue in Synechocystis sp. PCC 6803. Analysis of a slr1270 insertion deletion mutant and respective wild-type revealed that the mutant presents increased susceptibility to antibiotics. In addition, a detailed study of the exoproteome showed that Slr1270 mediates protein secretion. Among the protein substrates dependent on Slr1270 function, we found the S-layer structural component. Electron microscopy studies of the slr1270 mutant showed that the S-layer is indeed absent. The requirement of functional Slr1270 for protein secretion and drug resistance mechanisms suggests that Slr1270 plays a role similar to that described for TolC in other bacteria. Additional phenotypic traits could also be observed, including slower growth rates at low temperature, impairment in biofilm formation and increased activity of enzymes detoxifying reactive oxygen species. Furthermore, an increased capacity of outer membrane vesicles (OMVs) formation and release was also found in the slr1270 mutant, a feature that has not yet been observed in bacteria lacking TolC. This work highlights the marked physiological fitness that the TolC-like Slr1270 bestows to the photosynthetic model Synechocystis sp. PCC 6803 and presents a valuable model for studying OMVs formation and release.

Background: Orthogonal, synthetic control devices were developed for Penicillium chrysogenum, a model filamentous fungus and industrially relevant cell factory. In the synthetic transcription factor, the QF DNA-binding domain of the transcription factor of the quinic acid gene cluster of Neurospora crassa is fused to the VP16 activation domain. This synthetic transcription factor controls the expression of genes under a synthetic promoter containing quinic acid upstream activating sequence (QUAS) elements, where it binds. A gene cluster may demand an expression tuned individually for each gene, which is a great advantage provided by this system. Results: The control devices were characterized with respect to three of their main components: expression of the synthetic transcription factors, upstream activating sequences, and the affinity of the DNA binding domain of the transcription factor to the upstream activating domain. This resulted in synthetic expression devices, with an expression ranging from hardly detectable to a level similar to that of highest expressed native genes. The versatility of the control device was demonstrated by fluorescent reporters and its application was confirmed by synthetically controlling the production of penicillin. Conclusions: The characterization of the control devices in microbioreactors, proved to give excellent indications for how the devices function in production strains and conditions. We anticipate that these well-characterized and robustly performing control devices can be widely applied for the production of secondary metabolites and other compounds in filamentous fungi.