Secretion and directed evolution of unspecific peroxygenases in S. cerevisiae.

Saccharomyces cerevisiae / genetics Escherichia coli Mixed Function Oxygenases / genetics Plasmids / genetics
Golden Gate Protein engineering Split-GFP UPO Yeast

Journal

Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271

Informations de publication

Date de publication:
2023
Historique:
medline: 27 11 2023
pubmed: 18 11 2023
entrez: 17 11 2023
Statut: ppublish

Résumé

Yeast-based secretion systems are advantageous for engineering highly interesting enzymes that are not or barely producible in E. coli. The herein-presented production setup facilitates high-throughput screening as no cell lysis is required. All techniques are described in detail, with access to freely available online tools and all vectors have been made available on the non-profit plasmid repository AddGene. We describe the method for UPOs as a model enzyme, showcasing their secretion, detection, and evolution using S. cerevisiae. Additional material to transfer this to P. pastoris has been published by our group previously (Püllmann & Weissenborn, 2021).

Identifiants

DOI: 10.1016/bs.mie.2023.09.013 PMID: 37977733
pubmed: 37977733
pii: S0076-6879(23)00322-1
doi: 10.1016/bs.mie.2023.09.013
pii:
doi:

Substances chimiques

peroxygenase EC 1.14.-
Mixed Function Oxygenases EC 1.-

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

267-306

Informations de copyright

Copyright © 2023. Published by Elsevier Inc.

Auteurs

Niklas Dietz (N)

Institute of Chemistry, Martin-Luther-University Halle-Wittenberg Weinbergweg 22, Halle (Saale), Germany.

Li Wan (L)

Institute of Chemistry, Martin-Luther-University Halle-Wittenberg Weinbergweg 22, Halle (Saale), Germany.

Judith Münch (J)

Institute of Chemistry, Martin-Luther-University Halle-Wittenberg Weinbergweg 22, Halle (Saale), Germany.

Martin J Weissenborn (MJ)

Institute of Chemistry, Martin-Luther-University Halle-Wittenberg Weinbergweg 22, Halle (Saale), Germany. Electronic address: martin.weissenborn@chemie.uni-halle.de.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Champignons › Levures › Saccharomyces › Saccharomyces cerevisiae › Secretion and directed evolution of unspecific...
questionsmedicales.fr › Bactéries › Proteobacteria › Gammaproteobacteria › Enterobacteriaceae › Escherichia › Escherichia coli › Secretion and directed evolution of unspecific...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Oxidoreductases › Oxygénases › Mixed function oxygenases › Secretion and directed evolution of unspecific...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Plasmides › Secretion and directed evolution of unspecific...