Analysis of proteomic changes in colored mutants of Xanthophyllomyces dendrorhous (Phaffia rhodozyma)

Abstract

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.

Extending our tools and resources in the non-conventional industrial yeast Xanthophyllomyces dendrorhous through the application of metabolite profiling methodologies

“…TCA and high respiratory cycles are related with ROS production, which increase the astaxanthin biosynthesis in Xanthophyllomyces (Schroeder and Johnson 1993 ). The close relation of carotenoid production, the TCA cycle and stress response has been reported with differential protein expression between wild-type and mutant strains (Barbachano-Torres et al 2014 ).…”

Expression of Xanthophyllomyces dendrorhous cytochrome-P450 hydroxylase and reductase in Mucor circinelloides

“…Bands with same size indicate that the transformants maintain the introduced DNA in circular plasmids, but plasmid rearranges can be also presumed World J Microbiol Biotechnol Two genes are responsible for the conversion of b-carotene to astaxanthin in X. dendrorhous: the crtS encoding a cytochrome-P450 hydroxylase (astaxanthin synthase) is responsible for the addition of hydroxyl and keto groups to the b-ionone rings of b-carotene and the crtR encoding a cytochrome-P450 reductase, as electron donor, is required to the CrtS for addition of functional groups to the substrate (Á lvarez et al 2006;Ojima et al 2006;Alcaíno et al 2008). Overexpression of the crtS gene in X. dendrorhous resulted in higher level of astaxanthin production (Contreras et al 2013), while point mutations in the gene led to astaxanthin non-producing and b-carotene accumulating yellow mutants (Á lvarez et al 2006;Ojima et al 2006;Barbachano-Torres et al 2014). Deletion of the crtR gene was not lethal, suggesting an alternative electron donor in X. dendrorhous, but the transformants were not able to accumulate astaxanthin, indicating that the crtS and crtR genes are also necessary for the conversion of b-carotene to astaxanthin (Alcaíno et al 2008).…”

Proteomic analysis of Rhodotorula mucilaginosa: dealing with the issues of a non‐conventional yeast

“…Thus, the increase in the expression level of enzymes involved in the biosynthesis of cytosolic acetyl‐CoA is in accordance with the accumulation of carotenoids in stationary phase. Moreover, in accordance with the upregulation of enzymes involved in the stress response during carotenogenesis (Barbachano‐Torres et al ., ; Martinez‐Moya et al ., ), at the stationary phase of growth there was a slight but consistent increase in the expression levels of catalase and heat shock protein HSP70. According to Martinez‐Moya et al .…”