Evaluation of new L-amino acids triethanolammonium salts usability for controlling protease activity.

Twenty new triethanolammonium amino acid salts (TEA AA) have been prepared from triethanolammonium hydroxide and L-amino acids. The physicochemical properties of TEA AA depended on the applied amino acid. Five of the synthesised salts, i.e. mono- and bis-salts of L-glutamic acid, L-aspartic acid, and TEA salt of l-glutamine were solids with melting points between 127.32 °C to 171.51 °C. The other TEA AA exhibited glass transition temperatures from -68.45 °C for TEA Ser to -6.27 °C for TEA Trp and were assigned as amino acid ionic liquids (AAILs). The TEA His was characterised by the highest thermal stability, with an average temperature of 5 % weight loss at 186.4 °C, whereas the lowest stability was determined for TEA Asp (107.5 °C). The developed salts were tested as reaction medium additives for proteolytic enzymes (papain, subtilisin, bromelain). Most AAILs showed an inhibitory effect on tested proteases but with different mechanisms related to the enzyme substrate specificity and structural diversity. The TEA Ser was the most effective competitive inhibitor (Ki = 0.24 10-4 mol/L) for bromelain, while TEA Val uncompetitive inhibitor for papain (Ki = 0.25 10-4 mol/L). The developed TEA AA salts exhibit potential as enzyme-controlling agents for use in industrial processes.

Structural, Thermal, and Storage Stability of Rapana Thomasiana Hemocyanin in the Presence of Cholinium-Amino Acid-Based Ionic Liquids.

Novel biocompatible compounds that stabilize proteins in solution are in demand for biomedical and/or biotechnological applications. Here, we evaluated the effect of six ionic liquids, containing mono- or dicholinium [Chol]1or2 cation and anions of charged amino acids such as lysine [Lys], arginine [Arg], aspartic acid [Asp], or glutamic acid [Glu], on the structure, thermal, and storage stability of the Rapana thomasiana hemocyanin (RtH). RtH is a protein with huge biomedicinal potential due to its therapeutic, drug carrier, and adjuvant properties. Overall, the ionic liquids (ILs) induce changes in the secondary structure of RtH. However, the structure near the Cu-active site seems unaltered and the oxygen-binding capacity of the protein is preserved. The ILs showed weak antibacterial activity when tested against three Gram-negative and three Gram-positive bacterial strains. On the contrary, [Chol][Arg] and [Chol][Lys] exhibited high anti-biofilm activity against E. coli 25213 and S. aureus 29213 strains. In addition, the two ILs were able to protect RtH from chemical and microbiological degradation. Maintained or enhanced thermal stability of RtH was observed in the presence of all ILs tested, except for RtH-[Chol]2[Glu].

The application of amino acid ionic liquids as additives in the ultrasound-assisted extraction of plant material.

The aim of the present study was to determine the antioxidant activity of the aqueous extracts from Lycopodium clavatum, Cetraria islandica and Dipsacus fullonum obtained using aqueous solutions of ionic liquids by the ultrasound-assisted extraction (IL-UAE) method. Triethanolammonium salts [TEAH]+[AA]- of four amino acids of different hydrophobicity - isoleucine - Ile, methionine - Met, threonine - Thr and arginine - Arg, were chosen as ionic liquids, because they are based on natural, bio-renewable raw materials, such as amino acids and contain a pharmaceutically and cosmetically acceptable counterion of triethanolamine. Triethanolammonium salts were synthesized, identified by spectroscopic methods (NMR and FT-IR) and characterized by thermal methods (DSC and TGA). The 2.5% w/v aqueous solutions of triethanolammonium amino acid salts were used as the solvents in combination with ultrasound assisted extraction (UAE). The estimation of antioxidant properties was carried out using the DPPH, FRAP and CUPRAC assays. Total polyphenol content was measured using the reagent Folin-Ciocalteu. The results showed that the use of [TEAH]+[Thr]- or [TEAH]+[Met]- aqueous solutions increased the antioxidant activity of extracts in comparison to that achieved for extracts with pure water. The use of [TEAH]+[Thr]- as an additive for ultrasound-assisted extraction was characterized by obtaining plant extracts with the highest antioxidant potential, even 2.4-fold. The use of the AAIL-UAE method allowed obtaining higher amounts of polyphenols compared to pure water extracts, even 5.5-fold. The used method allowed the extraction of thermosensitive natural compounds, shortened the extraction time and lowered energy consumption.