Cloning, expression, and characterization of a recombinant xylanase from Bacillus sonorensis T6

Abstract

Xylanase is one of industrial enzymes with diverse applications including the paper-bleaching industry and feed additives. Here, a strain having xylanolytic activity and identified as Bacillus sonorensis T6 was isolated from soil. A secretory enzyme was identified by mass-spectrometry as a xylanase of glycosyl hydrolase family 11, with a molecular weight of 23.3 kDa. The xylanase gene of Bacillus sonorensis T6 was cloned and expressed in Escherichia coli (yielding an enzyme designated as rXynT6-E) and in Pichia pastoris (yielding rXynT6-P). The recombinant xylanases were found to have optimal activity at 47–55°C and pH 6.0–7.0. The recombinant xylanase expressed in P. pastoris has 40% higher thermal stability than that expressed in E. coli. The recombinant xylanases retained 100% of activity after 10 h incubation in the pH range 3–11 and 68% of activity after 1 h at pH 2.0. The xylanase activities of rXynT6-E and rXynT6-P under optimal conditions were 1030.2 and 873.8 U/mg, respectively. The good stability in a wide range of pH and moderate temperatures may make the xylanase from Bacillus sonorensis T6 useful for various biotechnological applications, e.g., as an enzyme additive in the feed industry.

Bioengineered xylanase from Misiones Argentina rainforest: A bakery enhancement approach

“…The results obtained from the evaluation of the optimal pH in this study are consistent with the pH ranges defined as optimal for xylanases in the literature, as the optimal pH for the enzyme was found to be 5.6 (Kiribayeva et al., 2022; Zhan et al., 2014) (Figure 3a). The recombinant xylanase exhibited an optimal temperature of 60°C (Figure 3 b).…”

“…The study results indicate that xylanase production increased by nearly 22% when Mg++ metal ions were added (Figure 1e), similar to the findings of Subajini and Vasanthy (2014) and Kiribayeva et al. (2022). In this regard, we can elucidate that magnesium acts as an enzymatic cofactor.…”

“…The inclusion of calcium and iron in the culture medium resulted in a decrease in xylanase activity (Figure 1f and 1g), mirroring the observations made by Kiribayeva et al. (2022). In this regard, we can argue that although both elements are crucial for various cellular functions, they may not be critical for the specific activity of xylanase in this context.…”

“…When evaluating the stability of the purified xylanolytic enzyme, it was observed that it retained 100% of its activity at 60 • C after 1 h of incubation and 97% after 2 h (Figure 3d), similar to our results. There are numerous reports in the literature where these values are comparable (Kiribayeva et al, 2022).…”

Lignocellulolytic Biocatalysts: The Main Players Involved in Multiple Biotechnological Processes for Biomass Valorization

“…Concerning the heterologous expression of lignocellulolytic enzymes, it is sought to express a functional lignocellulolytic system in order to allow non-lignocellulolytic microorganisms to hydrolyze and transform lignocellulosic biomass. The most commonly used non-lignocellulolytic microorganisms for this purpose are Zymomonas mobilis , Escherichia coli , Pichia pastoris , and Saccharomyces cerevisiae [ 277 , 368 , 369 , 370 , 371 ]. The production of recombinant lignocellulases may be the solution to limitations of high substrate cost and maintenance of the necessary conditions for these enzymes production, as well as more resistant and stable strains production and higher rates of enzyme production [ 372 , 373 , 374 , 375 ].…”