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Artículo

Functional and structural characterization of an endo-β-1,3-glucanase from Euglena gracilis

Calloni, Rodrigo DanielIcon ; Muchut, Robertino JoséIcon ; Garay, Alberto Sergio; Arias, Diego GustavoIcon ; Iglesias, Alberto AlvaroIcon ; Guerrero, Sergio AdrianIcon
Fecha de publicación: 05/2023
Editorial: Elsevier France-Editions Scientifiques Medicales Elsevier
Revista: Biochimie
ISSN: 0300-9084
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Bioquímica y Biología Molecular

Resumen

Endo-β-1,3-glucanases from several organisms have attracted much attention in recent years because of their capability for in vitro degrading β-1,3-glucan as a critical step for both biofuels production and short-chain oligosaccharides synthesis. In this study, we biochemically characterized a putative endo-β-1,3-glucanase (EgrGH64) belonging to the family GH64 from the single-cell protist Euglena gracilis. The gene coding for the enzyme was heterologously expressed in a prokaryotic expression system supplemented with 3% (v/v) ethanol to optimize the recombinant protein right folding. Thus, the produced enzyme was highly purified by immobilized-metal affinity and gel filtration chromatography. The enzymatic study demonstrated that EgrGH64 could hydrolyze laminarin (KM 23.5 mg ml−1,kcat 1.20 s−1) and also, but with less enzymatic efficiency, paramylon (KM 20.2 mg ml−1,kcat 0.23 ml mg−1 s−1). The major product of the hydrolysis of both substrates was laminaripentaose. The enzyme could also use ramified β-glucan from the baker's yeast cell wall as a substrate (KM 2.10 mg ml−1, kcat 0.88 ml mg−1 s−1). This latter result, combined with interfacial kinetic analysis evidenced a protein's greater efficiency for the yeast polysaccharide, and a higher number of hydrolysis sites in the β-1,3/β-1,6-glucan. Concurrently, the enzyme efficiently inhibited the fungal growth when used at 1.0 mg/mL (15.4 μM). This study contributes to assigning a correct function and determining the enzymatic specificity of EgrGH64, which emerges as a relevant biotechnological tool for processing β-glucans.
Palabras clave: EUGLENOIDS , GH64 PROTEIN , LAMINARIN , PARAMYLON
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/212055
URL: https://linkinghub.elsevier.com/retrieve/pii/S0300908422003418
DOI: http://dx.doi.org/10.1016/j.biochi.2022.12.016
Colecciones
Articulos(IAL)
Articulos de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Citación
Calloni, Rodrigo Daniel; Muchut, Robertino José; Garay, Alberto Sergio; Arias, Diego Gustavo; Iglesias, Alberto Alvaro; et al.; Functional and structural characterization of an endo-β-1,3-glucanase from Euglena gracilis; Elsevier France-Editions Scientifiques Medicales Elsevier; Biochimie; 208; 5-2023; 117-128
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