Xylanases are categorized into different family groups, two of which are glycoside hydrolases 10 (GH10) and 11 (GH11) families. These well-characterized xylanases demonstrate different modes of action in hydrolysis of xylans. Imitating certain types of microorganisms to produce bifunctional enzymes such as engineered xylanases has gained considerable attention among researchers. In this study, a recombinant chimeric enzyme (X11-10) was designed by fusing two thermostable xylanases through a peptide linker. The recombinant parental enzymes, xylanase 10 from fungus Bispora sp. MEY-1 (X10) and xylanase 11 from bacterium Thermobacillus xylanilyticus (X11), and their chimera were successfully expressed in Pichia pastoris (P. pastoris), purified, and characterized. Being active over a wide pH range, X11-10 chimera showed higher thermal stability, possessed a lower K_m, and a higher catalytic efficiency (k_cat/K_m) in comparison to the parental enzymes. Also, molecular dynamics simulation (MDS) of X11-10 revealed that its active site residues were free to interact with substrate. This novel chimeric xylanase may have potential applications in different industrial processes since it can substitute two separate enzymes and therefore minimize the production costs.

木聚糖酶分为不同的家族组，其中两个是糖苷水解酶 10 (GH10) 和 11 (GH11) 家族。这些充分表征的木聚糖酶在木聚糖水解中表现出不同的作用模式。模仿某些类型的微生物来生产双功能酶，例如工程木聚糖酶，已在研究人员中引起了相当大的关注。在本研究中，通过肽接头融合两种耐热木聚糖酶，设计了一种重组嵌合酶 (X11-10)。来自真菌Bispora sp.的重组亲本酶，木聚糖酶 10 。MEY-1（X10）和木聚糖酶从细菌11 Thermobacillus xylanilyticus（X11），和它们的嵌合体中成功表达巴斯德毕赤酵母（巴斯德毕赤酵母)、纯化和表征。与亲本酶相比，X11-10 嵌合体在较宽的 pH 范围内具有活性，表现出更高的热稳定性、更低的K _m和更高的催化效率 ( k _cat / K _m )。此外，X11-10 的分子动力学模拟 (MDS) 显示其活性位点残基可自由与底物相互作用。这种新型嵌合木聚糖酶可能在不同的工业过程中具有潜在应用，因为它可以替代两种不同的酶，从而最大限度地降低生产成本。