Thermostable Carbohydrate-Active Enzymes for High-Temperature Biomass Conversion: From Molecular Design to Process Integration
- 1College of Engineering and Applied Sciences, Nanjing University, Nanjing 210023, China
- 2School of Food and Pharmaceutical Engineering, Nanjing Normal University, Nanjing 210023, China
Abstract
High-temperature processing of carbohydrate-rich biomass, including starch liquefaction and lignocellulose saccharification, can improve sugar yields, reduce contamination risks, and facilitate integration with thermochemical pretreatments. However, the operational stability of native carbohydrate-active enzymes (CAZymes) at 60–90 °C remains a major bottleneck for efficient and economically viable biorefineries, and existing reviews rarely integrate multiscale molecular mechanisms with process-relevant engineering strategies. This review systematically examines the multiscale basis of thermostability in CAZymes—covering core hydrophobic packing, electrostatic and metal-mediated networks, disulfide bonds, conformational dynamics, and modular architectures involving catalytic domains, linkers, and carbohydrate-binding modules—and directly links these features to enzyme longevity under high-solids, inhibitor-rich conditions. We then critically compare key engineering strategies, including structure-guided rational design, semi-rational directed evolution, consensus design, ancestral sequence reconstruction, and machine learning-assisted workflows embedded in design–build–test–learn (DBTL) cycles, highlighting their respective strengths, limitations, and complementarity. Representative case studies demonstrate that engineered thermophilic α-amylases and cellulases achieving ΔTm improvements of approximately 10–20 °C can sustain >80–85% residual activity after prolonged exposure at process-relevant temperatures, translating into tangible benefits such as 15–30% reductions in enzyme dosing and measurable decreases in steam consumption, alongside higher sugar titers. By bridging molecular determinants, engineering paradigms, and quantitative process performance, this review provides a pragmatic roadmap for deploying thermostable CAZymes as robust biocatalysts in sustainable high-temperature biomass conversion.
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BibTeX
@article{hu2026thermostable,
title = {{Thermostable Carbohydrate-Active Enzymes for High-Temperature Biomass Conversion: From Molecular Design to Process Integration}},
author = {Yueli Hu and Xi Yang and Tianqi Wang and Mingshu Zheng and Zhenghao Jiang and Xinyue Dou and Xinkun Ren and Jiaxin Wu},
journal = {International journal of molecular sciences},
year = {2026},
month = sep,
doi = {10.3390/ijms27198788},
url = {https://europepmc.org/article/PMC/PMC13665343},
note = {CC BY},
}
RIS
TY - JOUR
TI - Thermostable Carbohydrate-Active Enzymes for High-Temperature Biomass Conversion: From Molecular Design to Process Integration
AU - Yueli Hu
AU - Xi Yang
AU - Tianqi Wang
AU - Mingshu Zheng
AU - Zhenghao Jiang
AU - Xinyue Dou
AU - Xinkun Ren
AU - Jiaxin Wu
PY - 2026
DA - 2026/09/30
JO - International journal of molecular sciences
DO - 10.3390/ijms27198788
UR - https://europepmc.org/article/PMC/PMC13665343
AB - High-temperature processing of carbohydrate-rich biomass, including starch liquefaction and lignocellulose saccharification, can improve sugar yields, reduce contamination risks, and facilitate integration with thermochemical pretreatments. However, the operational stability of native carbohydrate-active enzymes (CAZymes) at 60–90 °C remains a major bottleneck for efficient and economically viable biorefineries, and existing reviews rarely integrate multiscale molecular mechanisms with process-relevant engineering strategies. This review systematically examines the multiscale basis of thermostability in CAZymes—covering core hydrophobic packing, electrostatic and metal-mediated networks, disulfide bonds, conformational dynamics, and modular architectures involving catalytic domains, linkers, and carbohydrate-binding modules—and directly links these features to enzyme longevity under high-solids, inhibitor-rich conditions. We then critically compare key engineering strategies, including structure-guided rational design, semi-rational directed evolution, consensus design, ancestral sequence reconstruction, and machine learning-assisted workflows embedded in design–build–test–learn (DBTL) cycles, highlighting their respective strengths, limitations, and complementarity. Representative case studies demonstrate that engineered thermophilic α-amylases and cellulases achieving ΔTm improvements of approximately 10–20 °C can sustain >80–85% residual activity after prolonged exposure at process-relevant temperatures, translating into tangible benefits such as 15–30% reductions in enzyme dosing and measurable decreases in steam consumption, alongside higher sugar titers. By bridging molecular determinants, engineering paradigms, and quantitative process performance, this review provides a pragmatic roadmap for deploying thermostable CAZymes as robust biocatalysts in sustainable high-temperature biomass conversion.
SN - 1422-0067
ER -