ArticleScientific reports2025
Identification of improved signal peptides for heterologous expression in Saccharomyces using a screen that exploits Gaussia luciferase.
Article in Scientific reports, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Empowering Precision C-H Functionalization: Advances in Peroxygenase Engineering.Angewandte Chemie (International ed. in English) · 2026Review
- N-Glycosylation Influences the Heterologous Expression of an Unspecific Peroxygenase From Marasmius rotula in Saccharomyces cerevisiae.Microbial biotechnology · 2026Article
- Enhanced secretion of an Agrocybe aegerita peroxygenase variant in K. phaffii using the native signal peptide.AMB Express · 2026Article
- Biological parts in yeast synthetic biology: from regulatory elements to predictive design platforms.FEMS yeast research · 2026Review
- Optimizing Yeast Surface-Displayed Unspecific Peroxygenase Production for Sustainable Biocatalysis.Bioengineering (Basel, Switzerland) · 2025Article
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3 authors.
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Abstract
A high-throughput and sensitive screen for the improved expression of gene targets in Saccharomyces cerevisiae is described that is based upon the activity of the luciferase from Gaussia princeps. Using the Unspecific Peroxygenase (UPO) from Agrocybe aegerita (AaeUPO) as a model protein, improvements in expression, effected through error-prone PCR-based mutation within the signal peptide (SP) domain, can be detected using fusion of the target to Gaussia luciferase encoded downstream of the first folded domain of the AaeUPO protein and luminescent assay of expression supernatants. In this way, previously undiscovered mutations within the SP of AaeUPO that improve expression were revealed, and then applied to the expression of full-length AaeUPO in S. cerevisiae. The system was validated against control expression constructs that were well or poorly expressed and indicated a 13.9-fold improvement in expression for the best mutant over the wild-type SP sequence. It is envisaged that this protocol may be applied generally to the high-throughput detection of improved expression in S. cerevisiae for constructs that are engineered using directed evolution techniques.
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