Algal polysaccharides (extracellular polysaccharides) and carbon nanotubes (CNTs) were adsorbed on dioctadecyldimethylammonium bromide Langmuir monolayers to serve as a matrix for the incorporation of urease. The physicochemical properties of the supramolecular system as a monolayer at the air-water interface were investigated by surface pressure-area isotherms, surface potential-area isotherms, interfacial shear rheology, vibrational spectroscopy, and Brewster angle microscopy. The floating monolayers were transferred to hydrophilic solid supports, quartz, mica, or capacitive electrolyte-insulator-semiconductor (EIS) devices, through the Langmuir-Blodgett (LB) technique, forming mixed films, which were investigated by quartz crystal microbalance, fluorescence spectroscopy, and field emission gun scanning electron microscopy. The enzyme activity was studied with UV-vis spectroscopy, and the feasibility of the thin film as a urea sensor was essayed in an EIS sensor device. The presence of CNT in the enzyme-lipid LB film not only tuned the catalytic activity of urease but also helped to conserve its enzyme activity. Viability as a urease sensor was demonstrated with capacitance-voltage and constant capacitance measurements, exhibiting regular and distinctive output signals over all concentrations used in this work. These results are related to the synergism between the compounds on the active layer, leading to a surface morphology that allowed fast analyte diffusion owing to an adequate molecular accommodation, which also preserved the urease activity. This work demonstrates the feasibility of employing LB films composed of lipids, CNT, algal polysaccharides, and enzymes as EIS devices for biosensing applications.
CyanoHABs (cyanobacterial harmful algal blooms) are blooms of cyanobacteria capable of producing cyanotoxins, a large group of secondary metabolites that are toxic to most eukaryotes. In this work, the main aim was to evaluate the presence of multiple genes from each of the clusters responsible for biosynthesis of cyanotoxins (cylindrospermopsin, microcystin and saxitoxin) in total DNA obtained from sixteen environmental water samples by PCR. Microcystin gene mcyE was amplified in all analyzed samples. Among the cylindrospermopsin genes analyzed, only the cyrC gene was amplified from DNA obtained from three of sixteen samples. Of the three different saxitoxin genes analyzed, sxtB and sxtI were present in four and three of the sixteen samples studied, respectively, and sxtA did not show any positive result. Based on our results, we suggest caution when using only one gene from the full clusters responsible for biosynthesis of cyanotoxins, given that it may not be sufficient to confirm or exclude the toxigenic potential of a sample.
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