Autodisplay of an endo-1,4-β-xylanase from Clostridium cellulovorans in Escherichia coli for xylans degradation.

Autor: Balderas Hernández VE; División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica (IPICYT), Camino a la Presa de San José 2055 Lomas 4ª. Sección, C.P. 78216, San Luis Potosí, Mexico., Salas-Montantes CJ; División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica (IPICYT), Camino a la Presa de San José 2055 Lomas 4ª. Sección, C.P. 78216, San Luis Potosí, Mexico., Barba-De la Rosa AP; División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica (IPICYT), Camino a la Presa de San José 2055 Lomas 4ª. Sección, C.P. 78216, San Luis Potosí, Mexico., De Leon-Rodriguez A; División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica (IPICYT), Camino a la Presa de San José 2055 Lomas 4ª. Sección, C.P. 78216, San Luis Potosí, Mexico. Electronic address: aleonr@me.com.
Jazyk: angličtina
Zdroj: Enzyme and microbial technology [Enzyme Microb Technol] 2021 Sep; Vol. 149, pp. 109834. Date of Electronic Publication: 2021 May 27.
DOI: 10.1016/j.enzmictec.2021.109834
Abstrakt: The goal of this work was the autodisplay of the endo β-1,4-xylanase (XynA) from Clostridium cellulovorans in Escherichia coli using the AIDA system to carry out whole-cell biocatalysis and hydrolysate xylans. For this, pAIDA-xynA vector containing a synthetic xynA gene was fused to the signal peptide of the toxin subunit B Vibro cholere (ctxB) and the auto-transporter of the synthetic aida gene, which encodes for the connector peptide and β-barrel of the auto-transporter (AT-AIDA). E. coli TOP10 cells were transformed and the biocatalyst was characterized using beechwood xylans as substrate. Optimal operational conditions were temperature of 55 °C and pH 6.5, and the Michaelis-Menten catalytic constants V max and K m were 149 U/g DCW and 6.01 mg/mL, respectively. Xylanase activity was inhibited by Cu 2+ , Zn 2+ and Hg 2+ as well as EDTA, detergents, and organic acids, and improved by Ca 2+ , Co 2+ and Mn 2+ ions. Ca 2+ ion strongly enhanced the xylanolytic activity up to 2.4-fold when 5 mM CaCl 2 were added. Also, Ca 2+ improved enzyme stability at 60 and 70 °C. Results suggest that pAIDA-xynA vector has the ability to express functional xylanase to perform whole-cell biocatalysis in order to hydrolysate xylans from hemicellulose feedstock.
(Copyright © 2021 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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