Enhancing purification of chitosanase from Metarhizium anisopliae by expanded bed adsorption chromatography using Doehlert design

Abstract The use of expanded bed adsorption with three different adsorbents Streamline (SP XL, DEAE and PHENYL) to purify the chitosanase from Metarhizium anisopliae strain with and without clarification using Doehlert design for the optimization of the experiments has been investigated. Chitosanases are enzymes that hydrolyze chitosan, a carbohydrate, resulting oligosaccharides that have many biological remarkable activities, such as anti-cancer, anti-HIV and anti-oxidants activities. The superficial velocity was evaluated in five levels, from 40 to 260 cm h −1 , while the type of adsorbent, a qualitative variable, was assessed at three levels, saving materials and time according to the Doehlert design. The results showed that it is possible to purify chitosanase from M . anisopliae by expanded bed adsorption. Chitosanase purification factor ranged from 2 to 3.1-fold and yield ranged from 27.7 to 45.3%. The highest enzyme activity in the elution was 0.06 U/mL values which were higher than or close to those reported. The molecular mass of chitosanase was about 45 kDa using SDS-PAGE.