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Title

Assembly of evolved ligninolytic genes in Saccharomyces cerevisiae

AuthorsGonzález-Pérez, David ; Alcalde Galeote, Miguel
KeywordsSaccharomyces cerevisiae
secretome
versatile peroxidase
laccase
white-rot fungi
ligninolytic oxidoreductases
directed evolution
Issue Date2014
PublisherTaylor & Francis
CitationBioengineered 5 (2014)
AbstractThe ligninolytic enzymatic consortium produced by white-rot fungi is one of the most efficient oxidative systems found in nature, with many potential applications that range from the production of 2nd generation biofuels to chemicals synthesis. In the current study, two high redox potential oxidoreductase fusion genes (laccase -Lac and versatile peroxidase -Vp-) that had been evolved in the laboratory were re-assembled in Saccharomyces cerevisiae. First, cell viability and secretion were assessed after co-transforming the Lac and Vp genes into yeast. Several expression cassettes were inserted in vivo into episomal bi-directional vectors in order to evaluate inducible promoter/terminator pairs of different strengths in an individual and combined manner. The synthetic white-rot yeast model harboring Vp(GAL1/CYC1)-Lac(GAL10/ADH1) displayed up to 1000 and 100 Units per L of peroxidase and laccase activity, respectively, representing a suitable point of departure for future synthetic biology studies. © 2014 Landes Bioscience. Do not distribute.
Publisher version (URL)https://doi.org/10.4161/bioe.29167
URIhttp://hdl.handle.net/10261/174177
DOI10.4161/bioe.29167
ISSN1949-1026
E-ISSN1949-1026
Appears in Collections:(ICP) Artículos
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