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<p>Lytic polysaccharide monooxygenases (LPMOs) are a class of copper-dependent oxidases capable of cleaving glycosidic bonds in crystalline polysaccharides through oxidative mechanisms, playing a pivotal role in biomass refining and green biomanufacturing. However, natural LPMOs still face technical bottlenecks such as insufficient electron transfer efficiency, limited catalytic activity, and poor stability under industrial reaction conditions. This review systematically summarizes the classification, structure, catalytic mechanisms, and diverse applications of LPMOs. It focuses on analyzing research progress in modifying LPMOs through protein engineering strategies such as rational design (e.g., disulfide bond engineering, active site optimization) and directed evolution. Furthermore, it summarizes current high-throughput technologies suitable for LPMO mutant screening. Finally, it outlines future development directions including AI-assisted design and cell-free expression systems, providing references for further enzyme engineering and industrial applications.</p>

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Protein engineering and high-throughput screening of lytic polysaccharide monooxygenases: strategies, challenges, and prospects for industrial applications

  • 期刊论文

  • 发表日期：

    2026年5月9日

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  • Ying Chen,
  • Wenjing Wang,
  • Sheng Chen,
  • Yong Feng,
  • Jia Zhao,
  • Guohui Li,
  • Huiqing Chen,
  • Yong Chen,
  • Huayou Chen

摘要

Lytic polysaccharide monooxygenases (LPMOs) are a class of copper-dependent oxidases capable of cleaving glycosidic bonds in crystalline polysaccharides through oxidative mechanisms, playing a pivotal role in biomass refining and green biomanufacturing. However, natural LPMOs still face technical bottlenecks such as insufficient electron transfer efficiency, limited catalytic activity, and poor stability under industrial reaction conditions. This review systematically summarizes the classification, structure, catalytic mechanisms, and diverse applications of LPMOs. It focuses on analyzing research progress in modifying LPMOs through protein engineering strategies such as rational design (e.g., disulfide bond engineering, active site optimization) and directed evolution. Furthermore, it summarizes current high-throughput technologies suitable for LPMO mutant screening. Finally, it outlines future development directions including AI-assisted design and cell-free expression systems, providing references for further enzyme engineering and industrial applications.