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The aim of our research was to develop an immobilization method for photocatalysts that is an alternative to the sol-gel or dip-coating methods and can be simply scaled up for technical applications. The investigated photocatalyst was TiO₂, which was electrochemically deposited onto a cathode made of stainless steel. This deposited film was photocatalytically active. In order to enhance the photoactivity of the TiO₂ film, commercially available P25 photocatalyst nanoparticles were occluded into the film. The effect of deposition current density as well as the amount of occluded nanoparticles on the photocatalytic activity and photoelectrochemical behavior was investigated. The photocatalytic activity was evaluated in a UV-LED reactor. The decomposition rate of toluene and cyclohexane in air was examined for all prepared stainless steel-photocatalyst composites. It was observed that deposits prepared with 5 g dm⁻³ of P25 in the deposition bath showed the best photocatalytic activity and highest photocurrent.
The proteolytic activity of three Pseudomonas aeruginosa strains, ATCC 27853 - a reference strain, and two clinical isolates was tested. The activity was examined after culturing the bacteria in two different growth media: the minimal M9 medium and rich Luria-Bertani broth (LB). Based on zymograms and protease activity specific assays, it was concluded that the reference strain produced three proteolytic enzymes in the LB medium: protease IV, elastase B and elastase A, while alkaline protease was only produced in the M9 medium. The clinical isolates of P. aeruginosa produced elastase B and alkaline protease when grown in the LB medium and the minimal M9 medium, respectively. PCR analysis confirmed the presence of both the lasB gene encoding elastase B and aprA coding for alkaline protease in the genomes of the three P. aeruginosa strains analyzed. The expression of these genes coding for two important P. aeruginosa virulence factors was dependent on the growth conditions in all the strains studied. The contribution of the extracellular proteinases to the virulence of P. aeruginosa strains used in this study was investigated using an insect model, the greater wax moth Galleria mellonella.
α-1,3-Glucanases (mutanases) are currently of great interest due to their potential use in the field of dental care. These enzymes have been reported in several bacteria, yeasts and fungi, but up to now, characterization of this family of proteins has been relatively poor. In this study, we identify and characterize a mutanase gene from Trichoderma harzianum CCM F-340. Sequence analysis, on the nucleotide and amino acid levels reveals that this α-1,3-glucanase is highly homologous to α-1,3-glucanases from T. harzianum isolate CBS 243.71 and T. asperellum CECT 20539. T. harzianum CCM F-340 mutanase is a 634-aa residue protein with a calculated molecular mass of 67.65 kDa, composed of two distinct, highly conserved domains (a long N-terminal catalytic domain and a short C-terminal polysaccharide-binding domain) separated by a less conserved Pro-Ser-Thr-rich linker region. The mutanase gene was expressed in an E. coli BL21(DE3) host, under the transcriptional control of T7 promoter. The purified enzyme migrated as a band of about 68 kDa after SDS-polyacrylamide gel electrophoresis, which coincided with the predicted size based on the amino acid sequence. Our data indicate that this enzyme is highly conserved in Trichoderma and can be produced in active form in such heterologous expression system.
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