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Cell-Free Noncanonical Redox Cofactor Systems

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Cell-Free Gene Expression

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2433))

Abstract

Noncanonical redox cofactor systems utilize nicotinamide adenine dinucleotide (phosphate), NAD(P)H, mimics to perform biotransformation reactions. Compared to systems utilizing native NAD(P)H, these noncanonical redox cofactors can offer decreased cost of cofactor supply, improved system activities, and can even supply reducing power directly to targeted reactions in complex biological environments. When these systems are operated in cell-free settings, the high level of user control afforded by direct access to the reaction system enables specific tuning of cofactor parameters, enzyme activity, and reaction progression to maximize system productivity. In this chapter, we will describe methods for constructing these cell-free noncanonical redox cofactor systems. Specifically, methods, design concepts, and system adaptation will be discussed for applying noncanonical redox cofactors to both purified protein-based and crude lysate-based biotransformation systems.

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, University of California Irvine, Irvine, CA, USA

    William B. Black & Han Li

Authors
  1. William B. Black
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  2. Han Li
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Corresponding author

Correspondence to Han Li .

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Editors and Affiliations

  1. Department of Chemical and Biological Engineering, Chemistry of Life Processes Institute, and Center for Synthetic Biology, Northwestern University, Evanston, IL, USA

    Ashty S. Karim

  2. Department of Chemical and Biological Engineering, Chemistry of Life Processes Institute, Center for Synthetic Biology, Robert H. Lurie Comprehensive Cancer Center, and Simpson Querrey Institute, Northwestern University, Evanston, IL, USA

    Michael C. Jewett

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Black, W.B., Li, H. (2022). Cell-Free Noncanonical Redox Cofactor Systems. In: Karim, A.S., Jewett, M.C. (eds) Cell-Free Gene Expression. Methods in Molecular Biology, vol 2433. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1998-8_11

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  • DOI: https://doi.org/10.1007/978-1-0716-1998-8_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1997-1

  • Online ISBN: 978-1-0716-1998-8

  • eBook Packages: Springer Protocols

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