title: Functional characterization of thiolase-encoding genes from Xanthophyllomyces dendrorhous and their effects on carotenoid synthesis.
abstract: The basidiomycetous yeast Xanthophyllomyces dendrorhous has been described as a potential biofactory for terpenoid-derived compounds due to its ability to synthesize astaxanthin. Functional knowledge of the genes involved in terpenoid synthesis would create opportunities to enhance carotenoid production. A thiolase enzyme catalyzes the first step in terpenoid synthesis.
  
  
  
  Two potential thiolase-encoding genes were found in the yeast genome; bioinformatically, one was identified as an acetyl-CoA C-acetyltransferase (ERG10), and the other was identified as a 3-ketoacyl Co-A thiolase (POT1). Heterologous complementation assays in Saccharomyces cerevisiae showed that the ERG10 gene from X. dendrorhous could complement the lack of the endogenous ERG10 gene in S. cerevisiae, thereby allowing cellular growth and sterol synthesis. X. dendrorhous heterozygous mutants for each gene were created, and a homozygous POT1 mutant was also obtained. This mutant exhibited changes in pigment composition and higher ERG10 transcript levels than the wild type strain.
  
  
  
  The results support the notion that the ERG10 gene in X. dendrorhous is a functional acetyl-CoA C-acetyltransferase essential for the synthesis of mevalonate in yeast. The POT1 gene would encode a functional 3-ketoacyl Co-A thiolase that is non-essential for cell growth, but its mutation indirectly affects pigment production.
authors.affiliation: Departamento de Ciencias Ecológicas y Centro de Biotecnología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Ñuñoa, Santiago, Chile.
authors.forename: Nicole
authors.initials: N
authors.lastname: Werner
authors.affiliation: Departamento de Ciencias Ecológicas y Centro de Biotecnología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Ñuñoa, Santiago, Chile.
authors.forename: Melissa
authors.initials: M
authors.lastname: Gómez
authors.affiliation: Departamento de Ciencias Ecológicas y Centro de Biotecnología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Ñuñoa, Santiago, Chile.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Baeza
authors.affiliation: Departamento de Ciencias Ecológicas y Centro de Biotecnología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Ñuñoa, Santiago, Chile.
authors.forename: Víctor
authors.initials: V
authors.lastname: Cifuentes
authors.affiliation: Departamento de Ciencias Ecológicas y Centro de Biotecnología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Ñuñoa, Santiago, Chile. jalcainog@u.uchile.cl.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Alcaíno
mesh_terms: D000101:Acetyl-CoA C-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D000102:Acetyl-CoA C-Acyltransferase / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001487:Basidiomycota / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D053898:Biosynthetic Pathways; D002338:Carotenoids / Q000096:biosynthesis*; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D060847:Metabolic Engineering / Q000379:methods; D009154:Mutation; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D013261:Sterols / Q000096:biosynthesis; D013729:Terpenes / Q000378:metabolism; D024341:Xanthophylls / Q000378:metabolism
chemical_list: D004271:DNA, Fungal; D013261:Sterols; D013729:Terpenes; D024341:Xanthophylls; D002338:Carotenoids; C005948:astaxanthine; D000102:Acetyl-CoA C-Acyltransferase; D000101:Acetyl-CoA C-Acetyltransferase
keywords: Astaxanthin; Carotenoids; Functional complementation; Mevalonate; Sterols; Thiolase