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

Arauzo‐Aguilera, Klaudia, Saaranen, Mirva J., Robinson, Colin, Ruddock, Lloyd 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 (2). Article Number e1350. ISSN 2045-8827. (doi:10.1002/mbo3.1350) (KAR id:100388)

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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.

Item Type: Article
DOI/Identification number: 10.1002/mbo3.1350
Uncontrolled keywords: CyDisCo, disulfide bond, Escherichia coli, periplasm, signal peptide, Tat pathway
Subjects: Q Science
Q Science > QH Natural history > QH301 Biology
Divisions: Divisions > Division of Natural Sciences > Biosciences
SWORD Depositor: JISC Publications Router
Depositing User: JISC Publications Router
Date Deposited: 10 Mar 2023 14:29 UTC
Last Modified: 06 Jun 2023 14:56 UTC
Resource URI: (The current URI for this page, for reference purposes)
Robinson, Colin:
Ruddock, Lloyd W.:
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