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Improvement of a synthetic live bacterial therapeutic for phenylketonuria with biosensor-enabled enzyme engineering

Author

Listed:
  • Kristin J. Adolfsen

    (Zymergen Inc. (formerly enEvolv Inc.))

  • Isolde Callihan

    (Zymergen Inc. (formerly enEvolv Inc.))

  • Catherine E. Monahan

    (Synlogic Inc)

  • Per Jr. Greisen

    (Zymergen Inc. (formerly enEvolv Inc.)
    Novo Nordisk Research Center Seattle Inc)

  • James Spoonamore

    (Zymergen Inc. (formerly enEvolv Inc.))

  • Munira Momin

    (Synlogic Inc)

  • Lauren E. Fitch

    (Zymergen Inc. (formerly enEvolv Inc.))

  • Mary Joan Castillo

    (Synlogic Inc)

  • Lindong Weng

    (Zymergen Inc. (formerly enEvolv Inc.)
    Sana Biotechnology)

  • Lauren Renaud

    (Synlogic Inc)

  • Carl J. Weile

    (Zymergen Inc. (formerly enEvolv Inc.))

  • Jay H. Konieczka

    (Zymergen Inc. (formerly enEvolv Inc.))

  • Teodelinda Mirabella

    (Synlogic Inc)

  • Andres Abin-Fuentes

    (Synlogic Inc)

  • Adam G. Lawrence

    (Zymergen Inc. (formerly enEvolv Inc.))

  • Vincent M. Isabella

    (Synlogic Inc)

Abstract

In phenylketonuria (PKU) patients, a genetic defect in the enzyme phenylalanine hydroxylase (PAH) leads to elevated systemic phenylalanine (Phe), which can result in severe neurological impairment. As a treatment for PKU, Escherichia coli Nissle (EcN) strain SYNB1618 was developed under Synlogic’s Synthetic Biotic™ platform to degrade Phe from within the gastrointestinal (GI) tract. This clinical-stage engineered strain expresses the Phe-metabolizing enzyme phenylalanine ammonia lyase (PAL), catalyzing the deamination of Phe to the non-toxic product trans-cinnamate (TCA). In the present work, we generate a more potent EcN-based PKU strain through optimization of whole cell PAL activity, using biosensor-based high-throughput screening of mutant PAL libraries. A lead enzyme candidate from this screen is used in the construction of SYNB1934, a chromosomally integrated strain containing the additional Phe-metabolizing and biosafety features found in SYNB1618. Head-to-head, SYNB1934 demonstrates an approximate two-fold increase in in vivo PAL activity compared to SYNB1618.

Suggested Citation

  • Kristin J. Adolfsen & Isolde Callihan & Catherine E. Monahan & Per Jr. Greisen & James Spoonamore & Munira Momin & Lauren E. Fitch & Mary Joan Castillo & Lindong Weng & Lauren Renaud & Carl J. Weile &, 2021. "Improvement of a synthetic live bacterial therapeutic for phenylketonuria with biosensor-enabled enzyme engineering," Nature Communications, Nature, vol. 12(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-26524-0
    DOI: 10.1038/s41467-021-26524-0
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    References listed on IDEAS

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    1. Sarel J Fleishman & Andrew Leaver-Fay & Jacob E Corn & Eva-Maria Strauch & Sagar D Khare & Nobuyasu Koga & Justin Ashworth & Paul Murphy & Florian Richter & Gordon Lemmon & Jens Meiler & David Baker, 2011. "RosettaScripts: A Scripting Language Interface to the Rosetta Macromolecular Modeling Suite," PLOS ONE, Public Library of Science, vol. 6(6), pages 1-10, June.
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