Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast

17 p.-7 fig.

Detalhes bibliográficos
Autores: Aza, Pablo, Molpeceres, Gonzalo, Salas, Felipe de, Camarero, Susana
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2021
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/236326
Acesso em linha:http://hdl.handle.net/10261/236326
Access Level:Acceso aberto
Palavra-chave:Signal peptide
Synthetic design
Enzyme heterologous expression
Α-factor preproleader
Saccharomyces cerevisiae
Directed evolution
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spelling Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeastAza, PabloMolpeceres, GonzaloSalas, Felipe deCamarero, SusanaSignal peptideSynthetic designEnzyme heterologous expressionΑ-factor preproleaderSaccharomyces cerevisiaeDirected evolution17 p.-7 fig.Saccharomyces cerevisiae plays an important role in the heterologous expression of an array of proteins due to its easy manipulation, low requirements and ability for protein post-translational modifications. The implementation of the preproleader secretion signal of the α-factor mating pheromone from this yeast contributes to increase the production yields by targeting the foreign protein to the extracellular environment. The use of this signal peptide combined with enzyme-directed evolution allowed us to achieve the otherwise difficult functional expression of fungal laccases in S. cerevisiae, obtaining different evolved α-factor preproleader sequences that enhance laccase secretion. However, the design of a universal signal peptide to enhance the production of heterologous proteins in S. cerevisiae is a pending challenge. We describe here the optimisation of the α-factor preproleader to improve recombinant enzyme production in S. cerevisiae through two parallel engineering strategies: a bottom-up design over the native α-factor preproleader (αnat) and a top-down design over the fittest evolved signal peptide obtained in our lab (α9H2 leader). The goal was to analyse the effect of mutations accumulated in the signal sequence throughout iterations of directed evolution, or of other reported mutations, and their possible epistatic interactions. Both approaches agreed in the positive synergism of four mutations (Aα9D, Aα20T, Lα42S, Dα83E) contained in the final optimised leader (αOPT), which notably enhanced the secretion of several fungal oxidoreductases and hydrolases. Additionally, we suggest a guideline to further drive the heterologous production of a particular enzyme based on combinatorial saturation mutagenesis of positions 86th and 87th of the αOPT leader fused to the target protein.This work has been funded by the Spanish project BIO2017-86559-R, and the WoodZymes project funded by the Bio Based Industries Joint Undertaking (JU) under grant agreement No 792070. The JU receives support from the European Union’s Horizon 2020 research and innovation programme and the Bio Based Industries Consortium.Peer reviewedSpringer NatureMinisterio de Ciencia, Innovación y Universidades (España)Aza, Pablo [0000-0002-8703-8399]Molpeceres, Gonzalo [0000-0002-4366-9412]Salas, Felipe de [0000-0002-0057-0180]Camarero, Susana [0000-0002-2812-895X]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202120212021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/236326reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/BIO2017-86559-Rinfo:eu-repo/grantAgreement/EC/H2020/792070https://doi.org/10.1007/s00018-021-03793-ySíinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2363262026-05-22T06:33:51Z
dc.title.none.fl_str_mv Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast
title Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast
spellingShingle Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast
Aza, Pablo
Signal peptide
Synthetic design
Enzyme heterologous expression
Α-factor preproleader
Saccharomyces cerevisiae
Directed evolution
title_short Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast
title_full Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast
title_fullStr Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast
title_full_unstemmed Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast
title_sort Design of an improved universal signal peptide based on the α‑factor mating secretion signal for enzyme production in yeast
dc.creator.none.fl_str_mv Aza, Pablo
Molpeceres, Gonzalo
Salas, Felipe de
Camarero, Susana
author Aza, Pablo
author_facet Aza, Pablo
Molpeceres, Gonzalo
Salas, Felipe de
Camarero, Susana
author_role author
author2 Molpeceres, Gonzalo
Salas, Felipe de
Camarero, Susana
author2_role author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia, Innovación y Universidades (España)
Aza, Pablo [0000-0002-8703-8399]
Molpeceres, Gonzalo [0000-0002-4366-9412]
Salas, Felipe de [0000-0002-0057-0180]
Camarero, Susana [0000-0002-2812-895X]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Signal peptide
Synthetic design
Enzyme heterologous expression
Α-factor preproleader
Saccharomyces cerevisiae
Directed evolution
topic Signal peptide
Synthetic design
Enzyme heterologous expression
Α-factor preproleader
Saccharomyces cerevisiae
Directed evolution
description 17 p.-7 fig.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/236326
url http://hdl.handle.net/10261/236326
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/BIO2017-86559-R
info:eu-repo/grantAgreement/EC/H2020/792070
https://doi.org/10.1007/s00018-021-03793-y
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Springer Nature
publisher.none.fl_str_mv Springer Nature
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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