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| Main Authors: | , , , , , , , , , |
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| Format: | Artículo científico |
| Language: | en |
| Published: |
Journal of agricultural and food chemistry
2025
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| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/41339057/ |
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Table of Contents:
- Structure-Guided Subunit Interface Engineering to Improve the Catalytic Efficiency of Dimeric Enzymes of FAH Family. Liang, Bo Meng, Chenfei Wang, Qian Du, Yafei Luo, Yu Zhao, Jixiang Wu, Danni Liang, Yajing Lu, Xuefeng Yang, Jianming Protein Engineering Biocatalysis Hydrolases Molecular Dynamics Simulation Bacterial Proteins Protein Multimerization Hydro-Lyases Xylose Kinetics The subunit interface plays a substantial role in the structures and functions of oligomeric enzymes, yet targeted mutations remain difficult to predict. Here, we targeted 2-keto-3-deoxy-d-xylonate dehydratase (XylX), the rate-limiting catalyst in the Weimberg pathway for d-xylose catabolism and a member of the fumarylacetoacetate hydrolase (FAH) family, which forms a compact homodimer. Guided by its crystal structure, we engineered the dimer interface and obtained triple mutant L210A/P181Q/Q308A, which showed a 6.04-fold increase in catalytic efficiency. Molecular dynamics simulations revealed that moderate enhancement of intersubunit flexibility accelerates substrate binding. When the mutant was coupled with other Weimberg enzymes in a one-pot process, 77% of d-xylose was converted to 56.05 ± 0.39 g/L α-ketoglutaric acid within 6 h. Moreover, this strategy is also applicable to other dimeric enzymes within the FAH family. This study highlights a promising strategy for engineering dimeric enzymes with a higher catalytic efficiency for producing valuable chemicals.