A Highly Stable Biocatalyst Obtained from Covalent Immobilization of a Non-Commercial Cysteine Phytoprotease

Abstract

In this work, araujiain (enzymatic preparation obtained from the latex of Araujia hortorum fruits) was successfully immobilized on glyoxyl-agarose via multipoint covalent attachment. Thus, good efficiency of immobilization and high operational stability of immobilized enzyme were obtained. The activity of araujiain at alkaline pH was significantly improved after immobilization. In addition, immobilized araujiain also showed high activity and good stability, without significant loss in its activity, at temperatures between 37 and 60°C and in the presence of immiscible organic solvents. Immobilized araujiain also showed good performance in a mixture of 50% ethyl acetate in buffer, used for peptide synthesis, with better results than when the free enzyme was used as catalyst. These results indicate that immobilized araujiain via multipoint covalent attachment can be highly stabilized and this method might be used for practical applications of araujiain in hydrolytic and synthetic processes.

Estudio bioquímico y biológico de nuevos extractos proteicos de plantas alimentarias: Potencial aplicación como agentes anticoagulantes, hipoglucemiantes, antioxidantes y anti-infectivos naturales

“…La actividad de papaína fue registrada por el aumento de la absorbancia a 410 nm producto de la hidrólisis del sustrato PFLNA (L-piroglutamil-L-fenilalanina-L-leucina-p-nitroanilida), mediante medidas continuas durante 10 min a 37 ºC (Obregón et al, 2015). Los volúmenes fueron adaptados para la lectura en placa de 96 pocillos (Lector de placas Tecan Infinite 200 Pro) en un volumen final de 200 µl (Tabla 2).…”