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Stress-Tolerant, Recyclable, and Renewable Biocatalyst Platform Enabled by Engineered Bacterial Spores

Hui, Yue and Cui, Ziyu and Sim, Seunghyun (2022) Stress-Tolerant, Recyclable, and Renewable Biocatalyst Platform Enabled by Engineered Bacterial Spores. ACS Synthetic Biology, 11 (8). pp. 2857-2868. ISSN 2161-5063. doi:10.1021/acssynbio.2c00256.

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Here, we describe a stress-tolerant, recyclable, and renewable biocatalyst platform based on T7 RNA polymerase-enabled high-density protein display on bacterial spores (TIED). TIED uses high-level T7 RNA polymerase-driven expression of recombinant proteins specifically in sporulating cells to allow spontaneous assembly of recombinant fusion proteins on the Bacillus subtilis spore surface. TIED enables high loading density in the range of 106 to 107 recombinant enzymes per spore, robust catalytic activity of displayed enzymes comparable to the respective free enzymes, and enhanced kinetic stability of displayed enzymes in methanol and elevated temperatures. Furthermore, we demonstrate TIED enzymes to be not only recyclable but also fully renewable after the loss of activity through induction of germination and sporulation, enabling perpetual regeneration of these immobilized biocatalysts.

Item Type:Article
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URLURL TypeDescription
Sim, Seunghyun0000-0002-4232-5917
Additional Information:© 2022 American Chemical Society. Received 16 May 2022. Published online 25 July 2022. This work is supported by the Institute for Collaborative Biotechnologies through cooperative agreement W911NF-19-2-0026 from the U.S. Army Research Office and by start-up funds (S.S.) from the University of California, Irvine. The authors thank the Bacillus Genetic Stock Center and Professor Jeffrey Tabor for providing bacterial strains. The authors thank Professors David Tirrell and Jennifer Prescher for their helpful comments. Author Contributions. Y.H. and S.S. designed experiments, analyzed the data, and wrote the article. Y.H. and Z.C. performed the experiments and analyzed the data. The authors declare no competing financial interest.
Funding AgencyGrant Number
Army Research Office (ARO)W911NF-19-2-0026
University of California, IrvineUNSPECIFIED
Subject Keywords:Bacteria, Fluorescence, Genetics, Peptides and proteins, Protein dynamics
Issue or Number:8
Record Number:CaltechAUTHORS:20220726-998265000
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Official Citation:Stress-Tolerant, Recyclable, and Renewable Biocatalyst Platform Enabled by Engineered Bacterial Spores Yue Hui, Ziyu Cui, and Seunghyun Sim ACS Synthetic Biology Article ASAP DOI: 10.1021/acssynbio.2c00256
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:115871
Deposited By: George Porter
Deposited On:27 Jul 2022 16:53
Last Modified:12 Oct 2022 00:09

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