Isolation and properties of citolyitic protein from sea anemone Metridium senile

Glede na prgišče člankov objavljenih objavljenih pred štiridesetimi leti, morska vetrnica Metridium senile vsebuje citolitičen protein, ki ga inhibira holesterol. Prav tako naj bi citolitične proteine vsebovala morska vetrnica Alicia mirabilis, njihovo prisotnost nakazujejo medicinska poročila o poškodbah ob kontaktu z njo in nedavna diplomska naloga. Za oba proteina smo predvidevali, da spadata v naddružino MACPF/CDC. V magistrski nalogi smo uspeli delno izolirati in očistiti oba proteina in določili nekatere njune biofizikalne lastnosti. Oba proteina sta velika okoli 55 kDa, temperaturno labilna z Tm pri 65 °C. Optimalna pH vrednost hemolize je 5,5 za Metridium oziroma 7,0 za Alicio, medtem ko je aktivnost obeh inhibirana pri pH 8,0. S testi vezave na umetne lipidne vezikle nismo ugotovili velike specifičnosti za posamezne lipide. Pridobili smo tudi aminokislinski sekvenci N-konca obeh proteinov in cDNA sekvenco citolitičnega proteina iz M. senile.According to a handful of papers published more than forty years ago, sea anemone Metridium senile contains cytolytic protein that is inhibited by cholesterol. Similarly, sea anemone Alicia mirabilis contains cytolytic protein which is probably the cause of several stings upon the contact with the anemone, as described in several medical reports. Both proteins are putative members of MACPF/CDC superfamily. In our research, we managed to partly purify both proteins and to determine some of their biophysical characteristics. Both proteins have molecular masses of approximately 55 kDa and are thermally labile with Tm at 65 °C. Optimal pH values for haemolysis are 5,5 (Metridium) and 7,0 (Alicia) while pH 8,0 proved to be inhibitory for both. Experiments with artificial lipid vesicles did not show strong biding specificity for used lipids. Additionally, we acquired N-terminal amino acid sequence for both proteins and cDNA sequence of cytolytic protein from M. senile

Bio-Based Epoxy Adhesives with Lignin-Based Aromatic Monophenols Replacing Bisphenol A

A bio-epoxy surface adhesive for adherence of the metal component species to glass substrate with desirable adhesion strength, converted controlled removal upon request, and bio-based resource inclusion was developed. For the development of resin, three different lignin-based aromatic monophenols, guaiacol, cresol, and vanillin, were used in the chemical epoxidation reaction with epichlorohydrin. The forming transformation process was studied by viscoelasticity, in situ FTIR monitoring, and Raman. Unlike other hydroxyl phenyls, guaiacol showed successful epoxide production, and stability at room temperature. Optimization of epoxide synthesis was conducted by varying NaOH concentration or reaction time. The obtained product was characterized by nuclear magnetic resonance and viscosity measurements. For the production of adhesive, environmentally problematic bisphenol A (BPA) epoxy was partially substituted with the environmentally acceptable, optimized guaiacol-based epoxy at 20, 50, and 80 wt.%. Mechanics, rheological properties, and the possibility of adhered phase de-application were assessed on the bio-substitutes and compared to commercially available polyepoxides or polyurethanes. Considering our aim, the sample composed of 80 wt.% bio-based epoxy/20 wt.% BPA thermoset was demonstrated to be the most suitable among those analyzed, as it was characterized by low BPA, desired boundary area and recoverability using a 10 wt.% acetic acid solution under ultrasound