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Construction of metal–organic framework-based multienzyme system for l-tert-leucine production

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A Correction to this article was published on 02 August 2023

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Abstract

Chiral compounds are important drug intermediates that play a critical role in human life. Herein, we report a facile method to prepare multi-enzyme nano-devices with high catalytic activity and stability. The self-assemble molecular binders SpyCatcher and SpyTag were fused with leucine dehydrogenase and glucose dehydrogenase to produce sc-LeuDH (SpyCatcher-fused leucine dehydrogenase) and GDH-st (SpyTag-fused glucose dehydrogenase), respectively. After assembling, the cross-linked enzymes LeuDH-GDH were formed. The crosslinking enzyme has good pH stability and temperature stability. The coenzyme cycle constant of LeuDH-GDH was always higher than that of free double enzymes. The yield of l-tert-leucine synthesis by LeuDH-GDH was 0.47 times higher than that by free LeuDH and GDH. To further improve the enzyme performance, the cross-linked LeuDH-GDH was immobilized on zeolite imidazolate framework-8 (ZIF-8) via bionic mineralization, forming LeuDH-GDH @ZIF-8. The created co-immobilized enzymes showed even better pH stability and temperature stability than the cross-linked enzymes, and LeuDH-GDH@ZIF-8 retains 70% relative conversion rate in the first four reuses. In addition, the yield of LeuDH-GDH@ZIF-8 was 0.62 times higher than that of LeuDH-GDH, and 1.38 times higher than that of free double enzyme system. This work provides a novel method for developing multi-enzyme nano-device, and the ease of operation of this method is appealing for the construction of other multi-enzymes @MOF systems for the applications in the kinds of complex environment.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Change history

  • 03 August 2023

    The original online version of this article was revised: To update the given and the family name of the authors

  • 02 August 2023

    A Correction to this paper has been published: https://doi.org/10.1007/s00449-023-02916-y

References

  1. Gladiali S (2007) Guidelines and methodologies in asymmetric synthesis and catalysis. C R Chim 10(3):220–231

    Article  CAS  Google Scholar 

  2. Luo W et al (2020) Cloning and expression of a novel leucine dehydrogenase: characterization and l-tert-Leucine production. Front Bioeng Biotechnol 8:186

    Article  PubMed  PubMed Central  Google Scholar 

  3. Jiang W, Fang B (2020) Synthesizing chiral drug intermediates by biocatalysis. Appl Biochem Biotechnol 192(1):146–179

    Article  CAS  PubMed  Google Scholar 

  4. Solano DM et al (2012) Industrial biotransformations in the synthesis of building blocks leading to enantiopure drugs. Biores Technol 115:196–207

    Article  Google Scholar 

  5. Menzel A et al (2004) From enzymes to “designer bugs” in reductive amination: a new process for the synthesis of L-tert-leucine using a whole cell-catalyst. Eng Life Sci 4(6):573–576

    Article  CAS  Google Scholar 

  6. Bommarius AS, Schwarm M, Drauz K (1998) Biocatalysis to amino acid-based chiral pharmaceuticals—examples and perspectives. J Mol Catal B Enzym 5(1–4):1–11

    Article  CAS  Google Scholar 

  7. Young Hong E et al (2010) Asymmetric synthesis of L-tert-leucine and L-3-hydroxyadamantylglycine using branched chain aminotransferase. J Mol Catal B Enzym 66(1–2):228–233

    Article  Google Scholar 

  8. Jin J-Z, Chang D-L, Zhang J (2011) Discovery and application of new bacterial strains for asymmetric synthesis of l-tert-butyl leucine in high enantioselectivity. Appl Biochem Biotechnol 164(3):376–385

    Article  CAS  PubMed  Google Scholar 

  9. Liu W et al (2014) Efficient synthesis of l-tert-leucine through reductive amination using leucine dehydrogenase and formate dehydrogenase coexpressed in recombinant E. coli. Biochem Eng J 91:204–209

    Article  CAS  Google Scholar 

  10. Clive DL, Etkin N (1994) Synthesis of α-amino acids by addition of putative azido radicals to α-methoxy acrylonitriles derived from aldehydes and ketones. Tetrahedron Lett 35(16):2459–2462

    Article  CAS  Google Scholar 

  11. Boesten WH et al (2001) Asymmetric Strecker synthesis of α-amino acids via a crystallization-induced asymmetric transformation using (R)-phenylglycine amide as chiral auxiliary. Org Lett 3(8):1121–1124

    Article  CAS  PubMed  Google Scholar 

  12. Drauz K, Gröger H, May O (2012) Enzyme catalysis in organic synthesis, 3 volume set, vol 1. John Wiley & Sons, New Jersey

    Book  Google Scholar 

  13. Yang X et al (2016) High efficient co-expression of leucine dehydrogenase and glucose dehydrogenase in Escherichia coli. Acta Microbiol Sin 56(11):1709–1718

    CAS  Google Scholar 

  14. Shu-ting LI, Min W (2009) Research on conditions of l-tert-leucine biosynthesis by genetically engineered Escherichia coli. Pharm Biotechnol 16(3):202–206

    Google Scholar 

  15. Bülow L, Ljungcrantz P, Mosbach K (1985) Preparation of a soluble bifunctional enzyme by gene fusion. Bio/Technology 3(9):821–823

    Google Scholar 

  16. Fan Z et al (2009) Multimeric hemicellulases facilitate biomass conversion. Appl Environ Microbiol 75(6):1754–1757

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Yerabham AS et al (2013) Revisiting disrupted-in-schizophrenia 1 as a scaffold protein. Biol Chem 394(11):1425–1437

    Article  CAS  PubMed  Google Scholar 

  18. Hyeon JE, Jeon SD, Han SO (2013) Cellulosome-based, Clostridium-derived multi-functional enzyme complexes for advanced biotechnology tool development: advances and applications. Biotechnol Adv 31(6):936–944

    Article  CAS  PubMed  Google Scholar 

  19. Zakeri B et al (2012) Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proc Natl Acad Sci 109(12):E690–E697

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Hagan RM et al (2010) NMR spectroscopic and theoretical analysis of a spontaneously formed Lys-Asp isopeptide bond. Angew Chem 122(45):8599–8603

    Article  Google Scholar 

  21. Banerjee A, Howarth M (2018) Nanoteamwork: covalent protein assembly beyond duets towards protein ensembles and orchestras. Curr Opin Biotechnol 51:16–23

    Article  CAS  PubMed  Google Scholar 

  22. Reddington SC, Howarth M (2015) Secrets of a covalent interaction for biomaterials and biotechnology: SpyTag and SpyCatcher. Curr Opin Chem Biol 29:94–99

    Article  CAS  PubMed  Google Scholar 

  23. Liu D et al (2017) Topology engineering of proteins in vivo using genetically encoded, mechanically interlocking SpyX modules for enhanced stability. ACS Cent Sci 3(5):473–481

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Liu L et al (2018) Construction of intracellular self-assembled multienzyme complex by SpyTag/SpyCatcher to achieve efficient biosynthesis. China Biotechnol 38(7):75–82

    Google Scholar 

  25. Dordick JS (1992) Designing enzymes for use in organic solvents. Biotechnol Prog 8(4):259–267

    Article  CAS  PubMed  Google Scholar 

  26. Cantone S et al (2013) Efficient immobilisation of industrial biocatalysts: criteria and constraints for the selection of organic polymeric carriers and immobilisation methods. Chem Soc Rev 42(15):6262–6276

    Article  CAS  PubMed  Google Scholar 

  27. Bailey JB et al (2017) Synthetic modularity of protein–metal–organic frameworks. J Am Chem Soc 139(24):8160–8166

    Article  CAS  PubMed  Google Scholar 

  28. Pilgrim BS, Champness NR (2020) Metal-organic frameworks and metal-organic cages–a perspective. Chem Plus Chem 85(8):1842–1856

    CAS  PubMed  Google Scholar 

  29. Bai W et al (2019) Ultrathin 2D metal–organic framework (nanosheets and nanofilms)-based x D–2D hybrid nanostructures as biomimetic enzymes and supercapacitors. J Mater Chem A 7(15):9086–9098

    Article  CAS  Google Scholar 

  30. Chen GS et al (2020) Embedding functional biomacromolecules within peptide-directed metal-organic framework (MOF) nanoarchitectures enables activity enhancement. Angew Chem Int Edn 59(33):13947–13954

    Article  CAS  Google Scholar 

  31. Liang W et al (2021) Metal–organic framework-based enzyme biocomposites. Chem Rev 121(3):1077–1129

    Article  CAS  PubMed  Google Scholar 

  32. Liang K et al (2017) Biomimetic mineralization of metal–organic frameworks around polysaccharides. Chem Commun 53(7):1249–1252

    Article  CAS  Google Scholar 

  33. Gao X et al (2021) Hierarchically porous magnetic Fe3O4/Fe-MOF used as an effective platform for enzyme immobilization: a kinetic and thermodynamic study of structure-activity. Catal Sci Technol 11(7):2446–2455

    Article  CAS  Google Scholar 

  34. Liao F-S et al (2017) Shielding against unfolding by embedding enzymes in metal–organic frameworks via a de novo approach. J Am Chem Soc 139(19):6530–6533

    Article  CAS  PubMed  Google Scholar 

  35. Qi B, Luo J, Wan Y (2018) Immobilization of cellulase on a core-shell structured metal-organic framework composites: better inhibitors tolerance and easier recycling. Biores Technol 268:577–582

    Article  CAS  Google Scholar 

  36. Liang K et al (2015) Biomimetic mineralization of metal–organic frameworks as protective coatings for biomacromolecules. Nat Commun 6(1):1–8

    Article  Google Scholar 

  37. Akimbekov Z et al (2017) Experimental and theoretical evaluation of the stability of true MOF polymorphs explains their mechanochemical interconversions. J Am Chem Soc 139(23):7952–7957

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The researchers are grateful for the support given by the National Natural Science Foundation of China (21878154) and the China Postdoctoral Science Foundation (2021M691624).

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Authors and Affiliations

  1. School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210046, China

    Ru Wang, Jianyao Jia, Xue Liu, Yaru Chen, Qing Xu & Feng Xue

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  1. Ru Wang
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  2. Jianyao Jia
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  3. Xue Liu
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  4. Yaru Chen
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  5. Qing Xu
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  6. Feng Xue
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Correspondence to Qing Xu or Feng Xue.

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Wang, R., Jia, J., Liu, X. et al. Construction of metal–organic framework-based multienzyme system for l-tert-leucine production. Bioprocess Biosyst Eng 46, 1365–1373 (2023). https://doi.org/10.1007/s00449-023-02900-6

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