Computation-Aided Phylogeny-Oriented Engineering of β-Xylosidase: Modification of "Blades" to Enhance Stability and Activity for the Bioconversion of Hemicellulose to Produce Xylose

被引：3

作者：

Zhang, Chenchen ^{[1
]}

Gao, Wenjing ^{[1
]}

Song, Zhaolin ^{[1
]}

Dong, Mengjun ^{[1
]}

Lin, Huixin ^{[1
]}

Zhu, Gang ^{[1
]}

Lian, Mengka ^{[1
]}

Xiao, Yunjie ^{[2
]}

Lu, Fuping ^{[1
]}

Wang, Fenghua ^{[1
]}

Liu, Yihan ^{[1
]}

机构：

[1] Tianjin Univ Sci & Technol, Coll Biotechnol, Key Lab Ind Fermentat Microbiol, Tianjin Key Lab Ind Microbiol, Tianjin 300457, Peoples R China

[2] Tianjin Univ, Sch Life Sci, Tianjin 300072, Peoples R China

来源：

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 2024年 / 72卷 / 05期

基金：

中国国家自然科学基金;

关键词：

beta-xylosidase; protein engineering; thermostability; catalyticefficiency; xylose; ANCESTRAL SEQUENCE RECONSTRUCTION; GEOBACILLUS-STEAROTHERMOPHILUS; LIGNOCELLULOSIC BIOMASS; MOLECULAR-DYNAMICS; DIRECTED EVOLUTION; WHEAT-STRAW; PRETREATMENT; THERMOSTABILITY; HYDROLYSIS; MECHANISM;

D O I：

10.1021/acs.jafc.3c08518

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

Hemicellulose is a highly abundant, ubiquitous, and renewable natural polysaccharide, widely present in agricultural and forestry residues. The enzymatic hydrolysis of hemicellulose has generally been accomplished using beta-xylosidases, but concomitantly increasing the stability and activity of these enzymes remains challenging. Here, we rationally engineered a beta-xylosidase from Bacillus clausii to enhance its stability by computation-aided design combining ancestral sequence reconstruction and structural analysis. The resulting combinatorial mutant rXYLO(M25I/S51L/S79E) exhibited highly improved robustness, with a 6.9-fold increase of the half-life at 60 degrees C, while also exhibiting improved pH stability, catalytic efficiency, and hydrolytic activity. Structural analysis demonstrated that additional interactions among the propeller blades in the catalytic module resulted in a much more compact protein structure and induced the rearrangement of the opposing catalytic pocket to mediate the observed improvement of activity. Our work provides a robust biocatalyst for the hydrolysis of agricultural waste to produce various high-value-added chemicals and biofuels.

引用

页码：2678 / 2688

页数：11