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

The goal of this work was the autodisplay of the endo beta-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 beta-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 degrees C and pH 6.5, and the Michaelis-Menten catalytic constants Vmax and Km were 149 U/gDCW and 6.01 mg/mL, respectively. Xylanase activity was inhibited by Cu2+, Zn2+ and Hg2+ as well as EDTA, detergents, and organic acids, and improved by Ca2+, Co2+ and Mn2+ ions. Ca2+ ion strongly enhanced the xylanolytic activity up to 2.4-fold when 5 mM CaCl2 were added. Also, Ca2+ improved enzyme stability at 60 and 70 degrees 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.

Automated summary

This study aimed to achieve the autodisplay of the endo beta-1,4-xylanase (XynA) from Clostridium cellulovorans in Escherichia coli using the AIDA system for whole-cell biocatalysis. Optimal operational conditions were found to be at a temperature of 55 degrees C and pH 6.5, with xylanase activity inhibited by certain ions and compounds but improved by others, such as Ca2+, Co2+, and Mn2+. The results indicate that the pAIDA-xynA vector has the capability to express functional xylanase for whole-cell biocatalysis in hydrolysing xylans from hemicellulose feedstock.