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Synthetic Biology for Cell-Free Biosynthesis: Fundamentals of Designing Novel In Vitro Multi-Enzyme Reaction Networks

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Book cover Synthetic Biology – Metabolic Engineering

Part of the book series: Advances in Biochemical Engineering/Biotechnology ((ABE,volume 162))

Abstract

Cell-free biosynthesis in the form of in vitro multi-enzyme reaction networks or enzyme cascade reactions emerges as a promising tool to carry out complex catalysis in one-step, one-vessel settings. It combines the advantages of well-established in vitro biocatalysis with the power of multi-step in vivo pathways. Such cascades have been successfully applied to the synthesis of fine and bulk chemicals, monomers and complex polymers of chemical importance, and energy molecules from renewable resources as well as electricity. The scale of these initial attempts remains small, suggesting that more robust control of such systems and more efficient optimization are currently major bottlenecks. To this end, the very nature of enzyme cascade reactions as multi-membered systems requires novel approaches for implementation and optimization, some of which can be obtained from in vivo disciplines (such as pathway refactoring and DNA assembly), and some of which can be built on the unique, cell-free properties of cascade reactions (such as easy analytical access to all system intermediates to facilitate modeling).

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Acknowledgment

The authors wish to acknowledge financial support from the EU project ST-FLOW (#289326), and the Swiss National Science Foundation (NCCR Molecular Systems Engineering). G. M. was a fellow of Becas Chile-CONICYT.

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  1. Bioprocess Laboratory, Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058, Basel, Switzerland

    Gaspar Morgado, Daniel Gerngross, Tania M. Roberts & Sven Panke

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Correspondence to Sven Panke .

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  1. Department of Chemical and Biomolecular Engineering, University of Illinois, Urbana, Illinois, USA

    Huimin Zhao

  2. Institut für Bioprozess- und Biosystemtechnik, Technische Universitðt Hamburg-Harburg, Hamburg, Germany

    An-Ping Zeng

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Morgado, G., Gerngross, D., Roberts, T.M., Panke, S. (2016). Synthetic Biology for Cell-Free Biosynthesis: Fundamentals of Designing Novel In Vitro Multi-Enzyme Reaction Networks. In: Zhao, H., Zeng, AP. (eds) Synthetic Biology – Metabolic Engineering. Advances in Biochemical Engineering/Biotechnology, vol 162. Springer, Cham. https://doi.org/10.1007/10_2016_13

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