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Self-sustaining closed-loop multienzyme-mediated conversion of amines into alcohols in continuous reactions

Nature Catalysis volume 1, pages 452–459 (2018)Cite this article

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Abstract

The synthesis of alcohols from amine starting materials is an excellent yet challenging strategy for the preparation of pharmaceuticals and polymers. Here we developed a versatile, self-sustaining closed-loop multienzymatic platform for the biocatalytic synthesis of a large range of non-commercially available products in a continuous flow with excellent yields (80 to >99%), reaction times and optical purity of secondary alcohols (>99 enantiomeric excess). This process was also extended to the conversion of biogenic amines into high-value alcohols, such as the powerful antioxidant hydroxytyrosol, and the synthesis of enantiopure 2-arylpropanols via the dynamic kinetic resolution of commercially affordable racemic amines. The system exploits the in situ immobilization of transaminases and redox enzymes which were combined to cater for a fully automated, ultra-efficient synthetic platform with cofactor recycling, in-line recovery of benign by-products and recirculation of the aqueous media that contains the recycled cofactors in catalytic amounts, which increases the efficiency of the system by over 20-fold.

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Fig. 1: Ultra-efficient closed-loop concept.
Fig. 2: Flow system assembly for the synthesis of primary alcohols.
Fig. 3: Synthesis of high-value alcohols from biogenic amines.
Fig. 4: Preparation of (S)-2-phenylpropanol from (R,S)-2-phenylpropylamine.
Fig. 5: Multienzymatic cascade synthesis of enantiopure (S)-1-phenylethanols.
Fig. 6: Ultra-efficient multienzyme conversion of amines into alcohols.

References

  1. Bartoszewicz, A., Ahlsten, N. & Martin-Matute, B. Enantioselective synthesis of alcohols and amines by iridium-catalysed hydrogenation, transfer hydrogenation, and related processes. Chem. Eur. J. 19, 7274–7302 (2013).

    Article  CAS  PubMed  Google Scholar 

  2. Ahn, Y., Ko, S. B., Kim, M. J. & Park, J. Racemization catalysts for the dynamic kinetic resolution of alcohols and amines. Coord. Chem. Rev. 252, 647–658 (2008).

    Article  CAS  Google Scholar 

  3. Romano, D., Gandolfi, R., Guglielmetti, S. & Molinari, F. Enzymatic hydrolysis of capsaicins for the production of vanillylamine using ECB deacylase from Actinoplanes utahensis. Food Chem. 124, 1096–1098 (2011).

    Article  CAS  Google Scholar 

  4. Duarte, D. R., Castillo, E., Barzana, E. & Lopez-Munguia, A. Capsaicin hydrolysis by Candida antarctica lipase. Biotechol. Lett. 22, 1811–1814 (2000).

    Article  CAS  Google Scholar 

  5. Pouy, M. J., Stanley, L. M. & Hartwig, J. F. Enantioselective, iridium-catalysed monoallylation of ammonia. J. Am. Chem. Soc. 131, 11312–11313 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Imm, S., Bähn, S., Neubert, L., Neumann, H. & Beller, M. An efficient and general synthesis of primary amines by ruthenium-catalysed amination of secondary alcohols with ammonia. Angew. Chem. Int. Ed. 122, 8303–8306 (2010).

    Article  Google Scholar 

  7. Mutti, F., Knaus, T., Scrutton, N. S., Breuer, M. & Turner, N. J. Conversion of alcohols to enantiopure amines through dual-enzyme hydrogen-borrowing cascades. Science 349, 1525–1529 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Baumgarten, R. J. Aliphatic deaminations in organic synthesis. J. Chem. Educ. 43, 398–408 (1966).

    Article  CAS  Google Scholar 

  9. Rahman, S. M. A., Ohno, H., Maezaki, N., Iwata, C. & Tanaka, T. Total synthesis of (±)-scopadulin. Org. Lett. 2, 2893–2895 (2000).

    Article  CAS  PubMed  Google Scholar 

  10. Kanbara, Y., Abe, T., Fushimi, N. & Ikeno, T. Base-catalysed direct transformation of benzylamines into benzyl alcohols. Synlett 23, 706–710 (2012).

    Article  CAS  Google Scholar 

  11. Carlquist P. M., Gorwa M. F. & Weber, N. Stereoselective biosynthesis in microbial host cells. World Intellectual Property Organization patent 2014/0376376 A1 (2014).

  12. Khusnutdinova, J. R., Ben-David, Y. & Milstein, D. Direct deamination of primary amines by water to produce alcohols. Angew. Chem. Int. Ed. 52, 6269–6272 (2013).

    Article  CAS  Google Scholar 

  13. Sadri, A.S. & Fiksdahl, A. Stereoselective transformation of amines via chiral 2,4,6-triphenylpyridinium intermediates. Tetrahedron: Asymmetry 12, 1947–1951 (2001).

    Article  Google Scholar 

  14. White, E. H., Li, M. & Lu, S. N-nitrososulfamates: sources of carbonium ions in aqueous media and substrates in solid-state decompositions. J. Org. Chem. 57, 1252–1258 (1992).

    Article  CAS  Google Scholar 

  15. Elswort, J. D. & Roth, R. H. Dopamine synthesis, uptake, metabolism, and receptors: relevance to gene therapy of Parkinson's disease. Exp. Neurol. 144, 4–9 (1997).

    Article  Google Scholar 

  16. Bouton, A. A. & Eisenhofer, G. Catecholamine metabolism: from molecular understanding to clinical diagnosis and treatment. Adv. Pharmacol. 42, 273–292 (1998).

    Article  Google Scholar 

  17. Tamborini, L., Fernandes, P., Paradisi, F. & Molinari, F. Flow bioreactors as complementary tools for biocatalytic process intensification. Trends Biotechnol. 36, 73–88 (2018).

    Article  CAS  PubMed  Google Scholar 

  18. Britton, J. & Jamison, T. F. The assembly and use of continuous flow systems for chemical synthesis. Nat. Protoc. 12, 2423–2446 (2017).

    Article  CAS  PubMed  Google Scholar 

  19. Yuryev, R., Strompen, S. & Liese, A. Coupled chemo(enzymatic) reactions in continuous flow. Beilstein J. Org. Chem. 7, 1449–1467 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Ricca, E., Brucher, B. & Schrittwieser, J. H. Multi‐enzymatic cascade reactions: overview and perspectives. Adv. Synth. Catal. 353, 2239–2262 (2011).

    Article  CAS  Google Scholar 

  21. Santacoloma, P. A., Sin, G., Gernaey, K. V. & Woodley, J. M. Multienzyme-catalysed processes: next-generation biocatalysis. Org. Process Res. Dev. 15, 203–212 (2011).

    Article  CAS  Google Scholar 

  22. Gruber, P. et al. Fundamentals and applications of immobilised microfluidic enzymatic reactors. Chem. Technol. Biotechnol. 86, 325–334 (2011).

    Article  CAS  Google Scholar 

  23. Lentes, J. et al. Competitive and sustainable manufacturing by means of ultra-efficient factories in urban surroundings. Int. J. Prod. Res. 55, 480–491 (2017).

    Article  Google Scholar 

  24. Garcia-Galan, C., Berenguer-Murcia, Á., Fernandez-Lafuente, R. & Rodrigues, R. C. Potential of different enzyme immobilisation strategies to improve enzyme performance. Adv. Synth. Catal. 353, 2885–2904 (2011).

    Article  CAS  Google Scholar 

  25. Abaházi, E. et al. Covalently immobilised Trp60Cys mutant of ω-transaminase from Chromobacterium violaceum for kinetic resolution of racemic amines in batch and continuous-flow modes. Biochem. Eng. J. 132, 270–278 (2018).

    Article  CAS  Google Scholar 

  26. Savile, C. K. et al. Biocatalytic asymmetric synthesis of chiral amines from ketones applied to sitagliptin manufacture. Science 329, 305–309 (2010).

    Article  CAS  PubMed  Google Scholar 

  27. Monti, D., Ottolina, G., Carrea, G. & Riva, S. Redox reactions catalysed by isolated enzymes. Chem. Rev. 111, 4111–4140 (2011).

    Article  CAS  PubMed  Google Scholar 

  28. Contente, M. L., Molinari, F., Serra, I., Pinto, A. & Romano, D. Stereoselective enzymatic reduction of ethyl secodione: preparation of a key intermediate for the total synthesis of steroids. Eur. J. Org. Chem. 2016, 1260–1263 (2016).

    Article  CAS  Google Scholar 

  29. Gomm, A. & O’Reilly, E. Transaminases for chiral amine synthesis. Curr. Opin. Chem. Biol. 43, 106–112 (2018).

    Article  CAS  PubMed  Google Scholar 

  30. Liu, W. & Wang, P. Cofactor regeneration for sustainable enzymatic biosynthesis. Biotechnol. Adv. 4, 269–384 (2007).

    Google Scholar 

  31. Schmidt, N. G., Simon, R. C. & Kroutil, W. Biocatalytic asymmetric synthesis of optically pure aromatic propargylic amines employing ω‐transaminases. Adv. Synth. Catal. 357, 1815–1821 (2015).

    Article  CAS  Google Scholar 

  32. Börner, T. et al. Explaining operational instability of amine transaminases: substrate-induced inactivation mechanism and Influence of quaternary structure on enzyme–cofactor intermediate stability. ACS Catal. 7, 1259–1269 (2017).

    Article  CAS  Google Scholar 

  33. Contente, M. L., Dall’Oglio, F., Tamborini, L., Molinari, F. & Paradisi, F. Highly efficient oxidation of amines to aldehydes with flow‐based biocatalysis. ChemCatChem 9, 3843–3848 (2017).

    Article  CAS  Google Scholar 

  34. Quaglia, D. et al. His-tagged horse liver alcohol dehydrogenase: immobilisation and application in the bio-based enantioselective synthesis of (S)-arylpropanols. Process Biochem. 48, 810–818 (2013).

    Article  CAS  Google Scholar 

  35. Contente, M. L. et al. Stereoselective reduction of aromatic ketones by a new ketoreductase from Pichia glucozyma. Appl. Microbiol. Biotechnol. 100, 193–201 (2016).

    Article  CAS  PubMed  Google Scholar 

  36. Satoha, Y., Tajima, K., Munekata, M., Keasling, J. D. & Lee, T. S. Engineering of l-tyrosine oxidation in Escherichia coli and microbial production of hydroxytyrosol. Metab. Eng. 14, 603–610 (2012).

    Article  CAS  Google Scholar 

  37. Dubhashe, Y. R., Sawant, V. M. & Gaikar, V. G. Process intensification of continuous flow synthesis of tryptophol. Ind. Eng. Chem. Res. 57, 2787–2796 (2018).

    Article  CAS  Google Scholar 

  38. Maślewski, P., Wyrzykowski, D., Witwicki, M. & Dołęga, A. Histaminol and its complexes with copper (ii)—studies in solid state solution. Eur. J. Inorg. Chem. 12, 1399–1408 (2018).

    Article  CAS  Google Scholar 

  39. Bordiga, M., Travaglia, F., Locatelli, M., Arlorio, M. & Coisson, J. D. Histaminol: identification and HPLC analysis of a novel compound in wine. J. Agric. Food Chem. 58, 10202–10208 (2010).

    Article  CAS  PubMed  Google Scholar 

  40. Planchestainer, M. et al. Continuous flow biocatalysis: production and in-line purification of amines by immobilised transaminase from Halomonas elongata. Green. Chem. 19, 372–375 (2017).

    Article  CAS  Google Scholar 

  41. Lichman, B. R. et al. One-pot triangular chemoenzymatic cascades for the synthesis of chiral alkaloids from dopamine. Green. Chem. 17, 852–855 (2015).

    Article  CAS  Google Scholar 

  42. Orenes-Piñero, E., García-Carmona, F. & Sánchez-Ferrer, A. A new process for obtaining hydroxytirosol using transformed Escherichia coli whole cells with phenol hydroxylase from Geobacillus thermoglucosidasius. Food Chem. 139, 377–383 (2013).

    Article  CAS  PubMed  Google Scholar 

  43. Giacomini, D., Galletti, P., Quintavalla, A., Gucciardo, G. & Paradisi, F. Highly efficient asymmetric reduction of arylpropionic aldehydes by horse liver alcohol dehydrogenase through dynamic kinetic resolution. Chem. Commun. 39, 4038–4040 (2007).

    Article  CAS  Google Scholar 

  44. Falus, P. et al. A continuous-flow cascade reactor system for subtilisin A-catalysed dynamic kinetic resolution of N-tert-butyloxycarbonylphenylalanine ethyl thioester with benzylamine. Adv. Synth. Catal. 358, 1608–1617 (2016).

    Article  CAS  Google Scholar 

  45. Dall’Oglio, F. et al. Flow-based stereoselective reduction of ketones using an immobilised ketoreductase/glucose dehydrogenase mixed bed system. Catal. Commun. 93, 29–32 (2017).

    Article  CAS  Google Scholar 

  46. De Maria, P. D. & Hollmann, F. On the (un)greenness of biocatalysis: some challenging figures and some promising options. Front. Microbiol 6, 1–5 (2015).

    Google Scholar 

  47. Rodríguez, C. et al. Steric vs. electronic effects in the Lactobacillis brevis ADH-catalysed bioreduction of ketones. Org. Biomol. Chem. 12, 673–681 (2014).

    Article  PubMed  Google Scholar 

  48. Cerioli, L., Planchestainer, M., Cassidy, J., Tessaro, D. & Paradisi, F. Characterization of a novel amine transaminase from Halomonas elongata. J. Mol. Cat. B 120, 141–150 (2015).

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by the Biotechnology and Biological Sciences Research Council [grant no. BB/P002536/1]; and the authors thank Resindion S.R.L. for donating the Sepabeads EC-EP/S.

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  1. School of Chemistry, University of Nottingham, Nottingham, United Kingdom

    Martina L. Contente & Francesca Paradisi

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M.L.C. performed the experimental work and analysed the results. M.L.C. and F.P. conceived and designed the experiments. M.L.C. and F.P. co-wrote the manuscript.

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Correspondence to Francesca Paradisi.

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Supplementary Methods, Supplementary Tables 1–4, Supplementary Equations 1 and 2, Supplementary Figures 1–3, Supplementary Note 1, Supplementary References, NMR Spectra

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Contente, M.L., Paradisi, F. Self-sustaining closed-loop multienzyme-mediated conversion of amines into alcohols in continuous reactions. Nat Catal 1, 452–459 (2018). https://doi.org/10.1038/s41929-018-0082-9

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