Studies on the Zymogram Method for the Detection of Pectinolytic Activities Using CTAB

Zymogram analysis is a useful tool for the identification of several enzymes. The present study was undertaken to investigate the efficiency gains from the characterization of pectic enzymes on zymograms by staining of pectin-agarose overlays using cetyl trimethyl ammonium bromide also known as cetrimide or CTAB. The method is based on the fact that the enzymatic hydrolysis of the pectic substrates included in the agarose matrix gel inhibited their precipitation by CTAB, leading to the appearance of cleared zones in front of the pectin hydrolases and lyases. Conversely, esterases led to the increase of pectin precipitation. Fungal pectinolytic enzymes were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subjected to the zymogram detection technique, using two pectin substances, namely citrus pectin and polygalacturonic acid. Overall, the findings presented in the current study indicate that several elements (ions, salts, pH, temperature, chelators, and reducing agents) may significantly affect the results of zymogram analysis and can, therefore, be employed to enhance the discriminatory and operational potential of the analysis in terms of accurate discrimination between several pectinolytic activities involved and effective implementation of the purification procedures required in the process.     

Isolation and characterization of kurstakin and surfactin isoforms produced by Enterobacter cloacae C3 strain

In this work, the extraction, structural analysis, and identification as well as antimicrobial, anti‐adhesive, and antibiofilm activities of lipopeptides produced by Enterobacter cloacae C3 strain were studied. A combination of chromatographic and spectroscopic techniques offers opportunities for a better characterization of the biosurfactant structure. Thin layer chromatography (TLC) and HPLC for amino acid composition determination are used. Efficient spectroscopic techniques have been utilized for investigations on the biochemical structure of biosurfactants, such as Fourier transform infrared (FT‐IR) spectroscopy and mass spectrometry analysis. This is the first work describing the production of different isoforms belonging to kurstakin and surfactin families by E cloacae strain. Three kurstakin homologues differing by the fatty acid chain length from C₁₀ to C₁₂ were detected. The spectrum of lipopeptides belonging to surfactin family contains various isoforms differing by the fatty acid chain length as well as the amino acids at positions four and seven. Lipopeptide C3 extract exhibited important antibacterial activity against Gram‐positive and Gram‐negative bacteria, antifungal activity, and interesting anti‐adhesive and disruptive properties against biofilm formation by human pathogenic bacterial strains: Salmonella typhimurium, Klebsiella pneumoniae, Staphylococcus aureus, Bacillus cereus, and Candida albicans.     

Gamma radiolysis of erythrosine dye in aqueous solutions

Beryllium-7, mainly measured via γ-spectrometry, is used as a (natural) radiotracer for education and science. For activities?<?0.1 Bq and samples containing also longer-lived ¹⁰Be, accelerator mass spectrometry (AMS) is the method-of-choice. We demonstrate that ⁷Be and ¹⁰Be can be quantified at the Dresden AMS facility on the same prepared BeO. Detection limits (⁷Be) are?~?0.6 mBq. Samples as small as tens of millilitres of rainwater can be chemically processed (after acidification) within a few hours without expensive and slow ion exchange. Isobar (⁷Li) suppression by chemistry and AMS is sufficient to guarantee for an ultrasensitive, cheap, and fast detection method for ⁷Be allowing high sample throughput.

     

Synthesis and cation complexation of calix[4]azacrowns and a spirobiscalix[4]azacrown

The synthesis and complexation properties of three new derivatives, one spirobiscalix[4]azacrown (1) and two calix[4]azacrown (2 and 3), are reported. Complexation studies of the three ligands toward transition and heavy metal cations have been carried out and monitored by UV absorption spectrophotometry in acetonitrile. Mononuclear complexes were detected for all complexes, whereas binuclear species (M₂L) were also formed in the case of ligand 1 with Cu²⁺ and Pb²⁺. The extraction properties of 1, 2 and 3 toward Cu²⁺, Zn²⁺, Pb²⁺, Ag²⁺ and Cd²⁺ are also reported. The results showed that complexation is the main factor affecting extraction with ligands 2 and 3, while with ligand 1 it is not.