University of Oulu

Arauzo-Aguilera, K., Saaranen, M. J., Robinson, C., & Ruddock, L. W. (2023). Highly efficient export of a disulfide-bonded protein to the periplasm and medium by the Tat pathway using CyDisCo in Escherichia coli. MicrobiologyOpen, 12, e1350.

Highly efficient export of a disulfide-bonded protein to the periplasm and medium by the Tat pathway using CyDisCo in Escherichia coli

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Author: Arauzo-Aguilera, Klaudia1; Saaranen, Mirva J.2; Robinson, Colin1;
Organizations: 1School of Biosciences, University of Kent, Canterbury, UK
2Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 1.1 MB)
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Language: English
Published: John Wiley & Sons, 2023
Publish Date: 2023-09-11


High-value heterologous proteins produced in Escherichia coli that contain disulfide bonds are almost invariably targeted to the periplasm via the Sec pathway as it, among other advantages, enables disulfide bond formation and simplifies downstream processing. However, the Sec system cannot transport complex or rapidly folding proteins, as it only transports proteins in an unfolded state. The Tat system also transports proteins to the periplasm, and it has significant potential as an alternative means of recombinant protein production because it transports fully folded proteins. Most of the studies related to Tat secretion have used the well-studied TorA signal peptide that is Tat-specific, but this signal peptide also tends to induce degradation of the protein of interest, resulting in lower yields. This makes it difficult to use Tat in the industry. In this study, we show that a model disulfide bond-containing protein, YebF, can be exported to the periplasm and media at a very high level by the Tat pathway in a manner almost completely dependent on cytoplasmic disulfide formation, by other two putative Tat SPs: those of MdoD and AmiC. In contrast, the TorA SP exports YebF at a low level.

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Series: MicrobiologyOpen
ISSN: 2045-8827
ISSN-E: 2045-8827
ISSN-L: 2045-8827
Volume: 12
Issue: 2
Article number: e1350
DOI: 10.1002/mbo3.1350
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Funding: This work was funded by the People Programme (Marie Skłodowska-Curie Actions) of the European Union's Horizon 2020 Programme under REA grant agreement no. 813979 (SECRETERS).
EU Grant Number: (813979) Secreters - A new generation of microbial expression hosts and tools for the production of biotherapeutics and high-value enzymes
Copyright information: © 2023 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.