Using protein engineering to accelerate implementation of continuous biocatalysis for API synthesis
September 15-19, 2019
Biocatalysis is an invaluable tool for the synthesis of many active pharmaceutical ingredients1,2. Enzymatic reactions are frequently performed in batch, but continuous biocatalysis is gaining interest in industry3, not least because it would allow better integration of chemical and enzymatic reaction steps4. However, selecting a suitable reactor configuration for continuous biocatalysis is often challenging due to the common limitations of enzymes, such as poor kinetic constants towards industrially relevant substrates, substrate/product inhibition, the need for an aqueous reaction environment and/or pH control. For this reason, we have recently developed a tool5 that presents a variety of reactor configurations, combining continuously stirred tank reactors (CSTRs) and continuous plug-flow reactors (CPFRs), to overcome the aforementioned limitations and facilitate reactor selection. However, the continued development of protein engineering technologies6,7 has revealed one of the strongest advantages of biocatalysis over chemocatalysis, namely that the properties of the catalyst can be modified to remove limitations and better suit industrial production processes.8,9 In this presentation we highlight how protein engineering, guided by process-specific targets, can streamline process development. For example, reducing product inhibition by protein engineering could allow operation in a single CPFR where the possibility of complete conversion greatly simplifies downstream processing, improving the tolerance of enzymes towards organic solvents could allow water-free operation without mass transfer limitations between phases and improving pH tolerance could remove the need for acid/base addition to combat pH changes during operation. Such changes to the catalyst could greatly facilitate process development in an industry where reduction of time-to-market is often critical.
(1) Pollard, D. J.; Woodley, J. M. Trends Biotechnol. 2007, 25, 66–73.
(2) Devine, P. N.; Howard, R. M.; Kumar, R.; Thompson, M. P.; Truppo, M. D.; Turner, N. J. Nat. Rev. Chem. 2018, 2, 409–421.
(3) Tamborini, L.; Fernandes, P.; Paradisi, F.; Molinari, F. Trends Biotechnol. 2018, 36, 73–88.
(4) Rudroff, F.; Mihovilovic, M. D.; Gröger, H.; Snajdrova, R.; Iding, H.; Bornscheuer, U. T. Nat. Catal. 2018, 1, 12–22.
(5) Lindeque, R. M.; Woodley, J. M. Catalysts 2019, 9, 262.
(6) Arnold, F. H. Angew. Chem. Int. Ed. 2018, 57, 4143–4148.
(7) Yang, K. K.; Wu, Z.; Arnold, F. H. arXiv:1811.10775 2018.
(8) Woodley, J. M. Curr. Opin. Chem. Biol. 2013, 17, 310–316.
(9) Woodley, J. M. Phil. Trans. R. Soc. A 2017, 376.
Rowan Lindeque and John M. Woodley, "Using protein engineering to accelerate implementation of continuous biocatalysis for API synthesis" in "Enzyme Engineering XXV", Huimin Zhao, University of Illinois at Urbana-Champaign, USA John Wong, Pfizer, USA Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/enzyme_xxv/33