Stability of antiacanthain in aqueous organic monophasic systems

BERSI, GRISEL; VALLÉS, DIEGO; CANTERA, ANA; BARBERIS, SONIA

Estancia Grande - San Luis.

Congreso; XXXII Reunión Científica Anual de la Sociedad de Biología de Cuyo; 2014

Sociedad de Biología de Cuyo

From a biotechnological perspective, there are many advantajes of employing enzymes in organic media, such as high regioselectivity and stereoselectivity, minimal side- chain protection requeriments. However, the major drawback of using enzymes in organic solvents is their significantly reduced activity compared to that in buffer media. The aim of this work was to study the stability of proteolytic enzyme of fruit of Bromelia antiacantha Bertol, a plant that growns in Argentina, in monophasic organic media, for its application to the bioactive peptide synthesis. An experimental statistical design allowed to cluster different organic solvents, by its physical chemistry characteristics and to select one of each group. Antiacanthain stability in monophasic system (Tris-HCl (0.1M) pH 8 - organic solvent) was assayed by means of incubation of pre-purified crude extract in the reaction mixture (30:70; 50:50 and 70:30) during 24 h. Enzyme specific activity was determined using N-alpha-benzoyl-DL-arginine-p-nitroanilide (BApNA) as substrate. Antiacanthain in monophasic systems, which were formed by buffer Tris-HCl (pH 8) and a miscible organic solvent, showed low activities ranging between 0.1 and 8 UI/mg de protein. These values were lower than 32% of the residual proteolytic activity in buffer. These results showed that these media will not allow the bioactive peptide synthesis because of them cause inactivation of the antiacanthain.