Abstract
Immobilizing enzymes in a metal–organic framework (MOF) is an effective approach to improve their stability and reusability, but the small pore sizes of most MOFs exclude many industrially relevant enzymes. Here, we present a dynamic bond-mediated approach that enables enzyme encapsulation beyond pore size constraints. We constructed a series of mesoporous MOFs by integrating robust trivalent metal–carboxylate clusters with dynamic divalent metal–pyridyl units. Systematic variation of metal combinations and linker lengths precisely tunes the framework stability and dynamics, enabling reversible dissociation and reformation of metal–pyridyl bonds. These dynamic bonds function as molecular gates, permitting enzymes larger than the intrinsic pores to infiltrate while preserving framework integrity. The strategy was applied to encapsulate diverse enzymes, preserving high enzymatic activity and enhancing operational stability. Furthermore, it supports the co-immobilization of multi-enzyme systems, such as NahK and GlmU, for efficient cascade synthesis of high-value glycosylated donors.
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Data availability
Previously published protein structures used in this study were obtained from the Protein Data Bank (PDB) under the following accession codes: 3CP5 (Cyt C) 1W4Y (HRP) 7×32 [https://doi.org/10.2210/pdb7X32/pdb] (NTR) 1TRH (Lipase) 3VIK (BGL) 4OCO (NahK) 1FWY (GlmU) Crystallographic data for the structures reported in this Article have been deposited at the Cambridge Crystallographic Data Center, under deposition numbers CCDC 2489486 (Cr-L1-Cu), 2489487 (Cr-L1-Pd),2489488 (Fe-L1-Cu), and 2489489 (Cr-L3-Pd). Copies of the data can be obtained free of charge via https://www.ccdc.cam.ac.uk/structures/. All other data supporting the findings of this study are available within the article and its supplementary information. Source data are provided with this paper.
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Acknowledgments
This work was supported by the National Key Research and Development Program of China (2025YFA1511500; S.Y., Y.C.L.), the National Natural Science Foundation of China (223B2107, 22271141, T2541071; S.Y., Y.C.L., X.Z., M.Q.), and the Natural Science Foundation of Jiangsu Province (BK20250064, BK20240032; S.Y., Y.C.L.). We thank the staff of the BL17B beamline (https://cstr.cn/31129.02.NFPS.BL17B) at the National Facility for Protein Science in Shanghai (NFPS, https://cstr.cn/31129.02.NFPS), Shanghai Advanced Research Institute, Chinese Academy of Sciences, for their technical support in Single-crystal XRD/PXRD. The theoretical calculations were conducted using the computing facilities of the High-Performance Computing Center (HPCC) at Nanjing University.
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Y.C.L. and M.Q. contributed equally to this work. S.Y., J.-L.Z., and X.Z. conceived the original idea. Y.C.L. and M.Q. performed the synthesis. Y.C.L. and M.Q. performed the comprehensive structural characterization, property measurements, and data analysis. L.G. conducted the DFT calculations. Y.X.Z. performed the ICP-OES measurements. Y.Y. and J.Z. performed the CLSM measurements. Y.C.L., M.Q., S.Y., J.-L.Z., and X.Z. drafted the manuscript. All authors contributed to the revision of the manuscript.
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Li, Y., Qiao, M., Gao, L. et al. Dynamic bond-driven encapsulation of enzymes in metal–organic frameworks beyond pore size constraints. Nat Commun (2026). https://doi.org/10.1038/s41467-026-70249-x
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DOI: https://doi.org/10.1038/s41467-026-70249-x
