Web of Science: 30 citations, Scopus: 29 citations, Google Scholar: citations,
An improved secretion signal enhances the secretion of model proteins from Pichia pastoris
Barrero, Juan J. (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Casler, Jason C. (University of Chicago. Department of Molecular Genetics and Cell Biology)
Valero Barranco, Francisco (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Ferrer, Pau (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Glick, Benjamin S. (University of Chicago. Department of Molecular Genetics and Cell Biology)

Date: 2018
Abstract: Background: proteins can be secreted from a host organism with the aid of N-terminal secretion signals. The budding yeast Pichia pastoris (Komagataella sp. ) is widely employed to secrete proteins of academic and industrial interest. For this yeast, the most commonly used secretion signal is the N-terminal portion of pre-pro-α-factor from Saccharomyces cerevisiae. However, this secretion signal promotes posttranslational translocation into the endoplasmic reticulum (ER), so proteins that can fold in the cytosol may be inefficiently translocated and thus poorly secreted. In addition, if a protein self-associates, the α-factor pro region can potentially cause aggregation, thereby hampering export from the ER. This study addresses both limitations of the pre-pro-α-factor secretion signal. - Results: we engineered a hybrid secretion signal consisting of the S. cerevisiae Ost1 signal sequence, which promotes cotranslational translocation into the ER, followed by the α-factor pro region. Secretion and intracellular localization were assessed using as a model protein the tetrameric red fluorescent protein E2-Crimson. When paired with the α-factor pro region, the Ost1 signal sequence yielded much more efficient secretion than the α-factor signal sequence. Moreover, an allelic variant of the α-factor pro region reduced aggregation of the E2-Crimson construct in the ER. The resulting improved secretion signal enhanced secretion of E2-Crimson up to 20-fold compared to the levels obtained with the original α-factor secretion signal. Similar findings were obtained with the lipase BTL2, which exhibited 10-fold enhanced secretion with the improved secretion signal. - Conclusions: the improved secretion signal confers dramatic benefits for the secretion of certain proteins from P. pastoris. These benefits are likely to be most evident for proteins that can fold in the cytosol and for oligomeric proteins.
Grants: Ministerio de Economía y Competitividad CTQ2016-74959-R
Agència de Gestió d'Ajuts Universitaris i de Recerca 2014/SGR-452
Rights: Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original. Creative Commons
Language: Anglès
Document: Article ; recerca ; Versió publicada
Subject: Translocation ; Secretion ; Pichia pastoris ; Alpha-factor ; Ost1 ; Heterologous protein production ; Aggregation
Published in: Microbial cell factories, Vol. 17 (2018) , art. 161, ISSN 1475-2859

DOI: 10.1186/s12934-018-1009-5
PMID: 30314480

13 p, 1.4 MB

The record appears in these collections:
Articles > Research articles
Articles > Published articles

 Record created 2019-01-09, last modified 2022-01-27

   Favorit i Compartir