Neutrase Immobilization on coconut and cashew apple fibers for peptide synthesis in organic media

CHINELATE, GERLA; SIRO, JERÓNIMO; ADRIANO, WELLINGTON; MAMMARELLA, ENRIQUE; GONÇALVES, LUCIANA

Florencia

Congreso; Tenth International Conference on Chemical and Process Engineering; 2011

Italian Association of Chemical Engineering

The enzyme immobilization on insoluble supports allows not only the reuse of the protein but also to modulate the catalyst properties. Besides, the use of an insoluble enzyme derivative enables the operation of continuous processes. Neutrase, a bacterial endoprotease produced by B. subtilis presents considerable interest due to a wide variety of possible applications, for example, transformations of cheese whey proteins in high value food. Thus, the aim of this work will be developing a neutrase immobilization protocol using cashew bagasse and green coconut fiber for enzyme immobilization. Cashew bagasse will be prepared by alkaline treatment and acid to remove lignin and hemi cellulose, respectively. By contrast, green coconut fiber will be washed with distilled water and dried. Both supports will be activated with glutaraldehyde, glycidol and epichlorohydrin. Neutrase (5mg enzyme.g of support -1) in sodium borate buffer will be added to the activated support and kept under stirring at 25°C for 3 to 24h. Soluble and immobilized neutrase activities will be assessed via spectrophotometer analysis at 700 nm according to the TCA- Lowry assay. The derivatives will be analyzed as immobilization yield, coupling yield and stabilization factor at 60°C and compared to soluble enzyme.