Extraction and purification of diamine oxidase from marine coproducts

Abstract

Several coproducts with high added value are nowdays not valued. This study was proposed with the aim of enhancing marine coproduts especially from crustacean. Three species of crustacea, two species of shrimp (Parapenaeus longirostris and Penaeus kerathurus) as well as Squilla mantis were used in order to extract and purify the diamine oxidase enzyme (DAO). An essay was also done from a vegetarian source using prickly pear racket of Opuncia officinalis as specie. Concentration and fractionation were performed by the addition of ammonium sulphate, (NH4)2SO4 saturation in stages and cold centrifuged at 17000 rpm. Enzyme activity of DAO crude and DAO fractionation results were tested using substrate Putrescine and measured with a spectrophotometer at a wavelength of 340 nm. Gel chromatographic methods were used for the separation of proteins and a Size Exclusion Chromatography (SEC, HPLC) method was used for the purification of the enzyme. The characterization of the enzyme was done by SDSPAGE and on-chip Electrophoresis. The Fourier transform infrared spectroscopy (FTIR) spectrum was determined.