RESEARCH ARTICLE
Functional and Structural Characterization of a Novel Isoamylase from Ostreococcus tauri and Role of the N-Terminal Domain
Nicolas Hedín1, Julieta Barchiesi1, Diego F. Gomez-Casati1, *, María V. Busi1, *
Article Information
Identifiers and Pagination:
Year: 2020Volume: 14
First Page: 1
Last Page: 11
Publisher ID: TOBIOTJ-14-1
DOI: 10.2174/1874070702014010001
Article History:
Received Date: 04/11/2019Revision Received Date: 28/11/2019
Acceptance Date: 04/01/2020
Electronic publication date: 12/02/2020
Collection year: 2020
open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: (https://creativecommons.org/licenses/by/4.0/legalcode). This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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.
