A molecular toolkit for heterologous protein secretion across Bacteroides species.

Autor:	Yeh YH; Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA., Kelly VW; Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA., Pour RR; Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.; Present address: Perlumi, Berkeley, CA 94704, USA., Sirk SJ; Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.; Department of Biomedical and Translational Sciences, Carle Illinois College of Medicine, Urbana, IL 61801, USA.; Department of Microbiology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.; Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.; Cancer Center at Illinois, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.; Lead Contact.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2023 Dec 15. Date of Electronic Publication: 2023 Dec 15.
DOI:	10.1101/2023.12.14.571725
Abstrakt:	Bacteroides species are abundant and prevalent stably colonizing members of the human gut microbiota, making them a promising chassis for developing long-term interventions for chronic diseases. Engineering these bacteria as on-site production and delivery vehicles for biologic drugs or diagnostics, however, requires efficient heterologous protein secretion tools, which are currently lacking. To address this limitation, we systematically investigated methods to enable heterologous protein secretion in Bacteroides using both endogenous and exogenous secretion systems. Here, we report a collection of secretion carriers that can export functional proteins across multiple Bacteroides species at high titers. To understand the mechanistic drivers of Bacteroides secretion, we characterized signal peptide sequence features as well as post-secretion extracellular fate and cargo size limit of protein cargo. To increase titers and enable flexible control of protein secretion, we developed a strong, self-contained, inducible expression circuit. Finally, we validated the functionality of our secretion carriers in vivo in a mouse model. This toolkit should enable expanded development of long-term living therapeutic interventions for chronic gastrointestinal disease. Competing Interests: Declaration of interests Y.H.Y. and S.J.S. have filed a patent application on this work (PCT/US2023/083131). The authors declare no other competing interests.
Databáze:	MEDLINE
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