Zhang, C., Recacha, R., Ruddock, L. W., & Moilanen, A. (2021). Efficient soluble production of folded cat allergen Fel d 1 in Escherichia coli. Protein Expression and Purification, 180, 105809. https://doi.org/10.1016/j.pep.2020.105809
Efficient soluble production of folded cat allergen Fel d 1 in Escherichia coli
|Author:||Zhang, Chi1; Recacha, Rosario1; Ruddock, Lloyd W.1;|
1Faculty of Biochemistry and Molecular Medicine, University of Oulu, Aapistie 7, 90220, Oulu, Finland
|Online Access:||PDF Full Text (PDF, 4.5 MB)|
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe2021050729183
|Publish Date:|| 2021-05-07
The major cat allergen Fel d 1 is one of the most common and potent causes of animal related allergy. Medical treatment of cat allergy has relied on immunotherapy carried out with cat dander extract. This approach has been problematic, mainly due to inconsistent levels of the major allergen in the produced extracts. Recombinant DNA technology has been proposed as an alternative method to produce more consistent pharmaceuticals for immunotherapy and diagnostics of allergy. Current approaches to produce recombinant Fel d 1 (recFel d 1) in the cytoplasm of Escherichia coli have however resulted in protein folding deficiencies and insoluble inclusion body formation, requiring elaborate in vitro processing to acquire folded material. In this study, we introduce an efficient method for cytoplasmic production of recFel d 1 that utilizes eukaryotic folding factors to aid recFel d 1 to fold and be produced in the soluble fraction of E. coli. The solubly expressed recFel d 1 is shown by biophysical in vitro experiments to contain structural disulfides, is extremely stable, and has a sensitivity for methionine sulfoxidation. The latter is discussed in the context of functional relevance.
Protein expression & purification
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
1182 Biochemistry, cell and molecular biology
This work was supported by the Academy of Finland (grant numbers 266457 and 272573), and Biocenter Oulu.
|Academy of Finland Grant Number:||
266457 (Academy of Finland Funding decision)
272573 (Academy of Finland Funding decision)
© 2020 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).