Analysis of proteomic changes in colored mutants of Xanthophyllomyces dendrorhous (Phaffia rhodozyma)
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The yeast Xanthophyllomyces dendrorhous synthesizes astaxanthin as its most prevalent xanthophyll derivative. Comparisons between the protein profiles of mutant lines of this yeast can provide insight into the carotenogenic pathway. Differently colored mutants (red, orange, pink, yellow, and white) were obtained from this yeast species, and their protein profiles were determined using two-dimensional polyacrylamide gel electrophoresis (2DE). Individual proteins differentially expressed were identified using mass spectrometry. The red mutants hyperproduced total carotenoids (mainly astaxanthin), while in white and orange mutants, mutagenesis affected the phytoene dehydrogenase activity as indicated by the accumulation of phytoene. Inactivation of astaxanthin synthase after the mutagenic treatment was evident in β-carotene accumulating mutants. Differences in the proteomic profiles of wild-type X. dendrorhous and its colored mutants were demonstrated using 2DE. Of the total number of spots detected in each gel (297–417), 128 proteins were present in all strains. The red mutant showed the greatest number of matches with respect to the wild type (305 spots), while the white and yellow mutants, which had reduced concentrations of total carotenoids, presented the highest correlation coefficient (0.6) between each other. A number of differentially expressed proteins were sequenced, indicating that tricarboxylic acid cycle and stress response proteins are closely related to the carotenogenic process.
KeywordsCarotenoid biosynthesis Proteomic profile Secondary metabolism Stress response Tricarboxylic acid cycle
We thank Professors E. Cerdá-Olmedo and J. Ávalos and their working groups from the Genetic Department at Seville University for their technical support in mutant isolation and characterization. A Barbachano-Torres and LM Castelblanco-Matiz thank CONACYT-México for doctoral (172824 and 219320, respectively) fellowships.
- Alcaíno J, Barahona S, Carmona M, Lozano C, Marcoleta A, Niklitschek M, Sepúlveda D, Marcelo B (2008) Cloning of the cytochrome P450 reductase (crtR) gene and its involvement in the astaxanthin biosynthesis of X. dendrhorhous. BMC Microbiol 8(189):1–13Google Scholar
- Álvarez V, Rodríguez-Sáiz M, de la Fuente JL, Gudiña EJ, Godio RP, Martín JF, Barredo JL (2006) The crtS gene of Xanthophyllomyces dendrorhous encodes a novel cytochrome-P450 hydroxylase involved in the conversion of beta-carotene into astaxanthin and other xanthophylls. Fungal Genet Biol 43:261–272PubMedCrossRefGoogle Scholar
- Barbachano-Torres A, Ramos-Valdivia AC, Cerda-García-Rojas CM, Salgado-Rodríguez LM, Flores-Ortiz C, Ponce-Noyola T (2012) Carotenogenesis induction with hydrogen peroxide in Xanthophyllomyces dendrorhous colored mutants. In: Mendez-Vilas A (ed) Microbes in applied research: current advances and challenges. World Scientific Publishing Co Pvt. Ltd., Singapore, pp 598–602CrossRefGoogle Scholar
- Chávez-Cabrera C, Flores-Bustamante ZR, Marsch R, del Montes CM, Sánchez S, Cancino-Díaz JC, Flores-Cotera LB (2010) ATP-citrate lyase activity and carotenoid production in batch cultures of Phaffia rhodozyma under nitrogen-limited and nonlimited conditions. Appl Microbiol Biotechnol 85:1953–1960PubMedCrossRefGoogle Scholar
- Kocharin K (2013) Metabolic engineering of Saccharomyces cerevisiae for polyhydroxybutyrate production. Dissertation, Chalmers University of Technology, Göteborg, SwedenGoogle Scholar
- Long TV (2004) Process for production of carotenoids, xanthophylls and apo-carotenoids utilizing eukaryotic microorganisms. US PATENT 7563935 B2Google Scholar
- Ojima K, Breitenbach J, Visser H, Setoguchi Y, Tabata K, Hoshino T, van den Berg J, Sandmann G (2006) Cloning of the astaxanthin synthase gene from Xanthophyllomyces dendrorhous (Phaffia rhodozyma) and its assignment as a β-carotene 3-hydroxylase/4-ketolase. Mol Genet Genomics 275:148–158PubMedCrossRefGoogle Scholar
- Yamane Y, Higashida K, Nakashimada Y, Kakizono T, Nishio N (1997) Influence of oxygen and glucose on primary metabolism and astaxanthin production by Phaffia rhodozyma in batch and fed-batch cultures: kinetic and stoichiometric analysis. Appl Environ Microbiol 63:4471–4478PubMedCentralPubMedGoogle Scholar