Functional and structural characterization of a novel isoamylase from ostreococcus tauri and role of the n-terminal domain

Abstract

Background: The debranching starch enzymes, isoamylase 1 and 2 are well-conserved enzymes present in almost all the photosynthetic organisms. These enzymes are involved in the crystallization process of starch and are key components which remove misplaced α-1,6 ramifications on the final molecule. Aim: In this work, we performed a functional and structural study of a novel isoamylase from Ostreococcus tauri. Methods: We identified conserved amino acid residues possibly involved in catalysis. We also identified a region at the N-terminal end that resembles a Carbohydrate Binding Domain (CBM), which is more related to the family CBM48, but has no spatial conservation of the residues involved in carbohydrate binding. Results: The cloning, expression and biochemical characterization of this N-terminal region confirmed that it binds to polysaccharides, showing greater capacity for binding to amylopectin rather than total starch or amylose. Conclusion: This module could be a variant of the CBM48 family or it could be classified within a new CBM family.