Zanoni Jéssica de Araujo, Zilli Izabela Karolina Costa, Pretto Guilherme de Paula, Seixas Flavio Augusto Vicente, Lima Marcela Marques de Freitas, Lemos Eliana Gertrudes de Macedo, Gomes Eleni, Zazeri Gabriel, Bonilla-Rodriguez Gustavo Orlando
Graduate Program in Microbiology, Institute of Biosciences, Humanities and Exact Sciences, São Paulo State University, IBILCE-UNESP, São José do Rio Preto 15054-000, SP, Brazil.
Department of Technology, State University of Maringá (UEM), Umuarama 87020-900, PR, Brazil.
BioTech (Basel). 2025 Aug 18;14(3):62. doi: 10.3390/biotech14030062.
Industrial applications of xylanases in high-temperature settings are limited by enzyme instability. This study evaluated glycerol and phenolic compounds as modulators of the catalytic and structural properties of a recombinant endoxylanase (rMhXyn) expressed in . Glycerol (20% /) significantly improved thermostability (5-fold increase in half-life at 55 °C), decreased the activation energy for catalysis, and enhanced structural rigidity as evidenced by molecular dynamics simulations (reduced RMSD and Rg). In contrast, phenolic acids provided only short-term stabilization at moderate temperatures and did not confer structural benefits. Enzyme kinetics revealed that glycerol enhanced catalytic turnover (↑), while phenolic compounds modified both ' and cooperativity (Hill coefficient). Thermodynamic analysis supported glycerol's stabilizing effect, with increased ∆ and a positive shift in ∆. These results suggest glycerol as a superior stabilizer for rMhXyn in high-temperature bioprocesses such as lignocellulosic biomass hydrolysis. These findings highlight the potential of targeted additives to improve enzyme performance for biotechnological applications.
木聚糖酶在高温环境下的工业应用受到酶不稳定性的限制。本研究评估了甘油和酚类化合物作为在[具体表达宿主]中表达的重组内切木聚糖酶（rMhXyn）催化和结构特性调节剂的作用。甘油（20% /）显著提高了热稳定性（55°C下半衰期增加了5倍），降低了催化的活化能，并增强了结构刚性，分子动力学模拟证明了这一点（均方根偏差和回转半径减小）。相比之下，酚酸仅在中等温度下提供短期稳定性，且未带来结构上的益处。酶动力学表明甘油增强了催化周转（↑），而酚类化合物改变了Km和协同性（希尔系数）。热力学分析支持了甘油的稳定作用，表现为∆H增加和∆S正向偏移。这些结果表明，在木质纤维素生物质水解等高温度生物过程中，甘油是rMhXyn的一种优良稳定剂。这些发现突出了靶向添加剂改善酶在生物技术应用中性能的潜力。
