Proteomic analysis of the carotenogenic yeast Xanthophyllomyces dendrorhous

Martínez-Moya, Pilar; Watt, Stephen A.; Niehaus, Karsten; Alcaı́no, Jennifer; Baeza, Marcelo; Cifuentes, Víctor

doi:10.1186/1471-2180-11-131

Cited by 21 publications

(18 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MM supplemented with 2% of glucose or succinate as a carbon source was used for growth of the wild-type X. dendrorhous strain UCD 67–385 [ 17 ]. The main culture (250 ml) was inoculated with 2.5 ml of an exponential phase culture and grown at 22°C with constant swirling at 120 rpm.…”

Section: Methodsmentioning

confidence: 99%

Proteomic and metabolomic analysis of the carotenogenic yeast Xanthophyllomyces dendrorhous using different carbon sources

et al. 2015

Self Cite

View full text Add to dashboard Cite

BackgroundAstaxanthin is a potent antioxidant with increasing biotechnological interest. In Xanthophyllomyces dendrorhous, a natural source of this pigment, carotenogenesis is a complex process regulated through several mechanisms, including the carbon source. X. dendrorhous produces more astaxanthin when grown on a non-fermentable carbon source, while decreased astaxanthin production is observed in the presence of high glucose concentrations. In the present study, we used a comparative proteomic and metabolomic analysis to characterize the yeast response when cultured in minimal medium supplemented with glucose (fermentable) or succinate (non-fermentable).ResultsA total of 329 proteins were identified from the proteomic profiles, and most of these proteins were associated with carotenogenesis, lipid and carbohydrate metabolism, and redox and stress responses. The metabolite profiles revealed 92 metabolites primarily associated with glycolysis, the tricarboxylic acid cycle, amino acids, organic acids, sugars and phosphates. We determined the abundance of proteins and metabolites of the central pathways of yeast metabolism and examined the influence of these molecules on carotenogenesis.Similar to previous proteomic-stress response studies, we observed modulation of abundance from several redox, stress response, carbohydrate and lipid enzymes. Additionally, the accumulation of trehalose, absence of key ROS response enzymes, an increased abundance of the metabolites of the pentose phosphate pathway and tricarboxylic acid cycle suggested an association between the accumulation of astaxanthin and oxidative stress in the yeast. Moreover, we observed the increased abundance of late carotenogenesis enzymes during astaxanthin accumulation under succinate growth conditions.ConclusionsThe use of succinate as a carbon source in X. dendrorhous cultures increases the availability of acetyl-CoA for the astaxanthin production compared with glucose, likely reflecting the positive regulation of metabolic enzymes of the tricarboxylic acid and glyoxylate cycles. The high metabolite level generated in this pathway could increase the cellular respiration rate, producing reactive oxygen species, which induces carotenogenesis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1484-6) contains supplementary material, which is available to authorized users.

show abstract

Section: Methodsmentioning

confidence: 99%

Proteomic and metabolomic analysis of the carotenogenic yeast Xanthophyllomyces dendrorhous using different carbon sources

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…To correlate the effect of assimilation of different carbon sources on central metabolism, we analyzed the proteome and transcriptome of strains cyc8 Ϫ/Ϫ , tup1 Ϫ/Ϫ , and mig1 Ϫ/Ϫ , encoding principal transcriptional regulators. Glucose repression and derepression essentially concern genes involved in oxidative metabolism and the TCA cycle, genes coding for the metabolism of alternative carbon sources, or genes coding for gluconeogenesis (11)(12)(13)(14)(15)(16)(17)(18)(19)(20). The most common mechanisms regulating gene products involve a decrease in transcription or translation at the gene level or an increase in protein degradation at the protein level (12).…”

Section: Discussionmentioning

confidence: 99%

“…Previously, Martinez-Moya and collaborators carried out proteomic studies in which they analyzed protein profiles present before and during the induction of carotenogenesis with glucose or succinate carbon sources. In those studies, differentially abundant proteins (DAPs) were identified by two-dimensional polyacrylamide gel electrophoresis (PAGE) followed by mass spectrometry (MS), and the data were corre-lated with the metabolomic profiles of central metabolic pathways (14,15). These works showed significant differences in the protein profiles related to the time of analysis and carbon source employed.…”

mentioning

confidence: 99%

Convergence between Regulation of Carbon Utilization and Catabolic Repression in Xanthophyllomyces dendrorhous

Martínez-Moya

Campusano

Córdova

et al. 2020

mSphere

Self Cite

View full text Add to dashboard Cite

Xanthophyllomyces dendrorhous is a carotenogenic yeast with a singular metabolic capacity to produce astaxanthin, a valuable antioxidant pigment. This yeast can assimilate several carbon sources and sustain fermentation even under aerobic conditions. Since astaxanthin biosynthesis is affected by the carbon source, the study of carotenogenesis regulatory mechanisms is key for improving astaxanthin yield in X. dendrorhous. This study aimed to elucidate the regulation of the metabolism of different carbon sources and the phenomenon of catabolic repression in this yeast. To this end, protein and transcript levels were quantified by iTRAQ (isobaric tags for relative and absolute quantification) and transcriptomic sequencing (RNA-seq) in the wild-type strain under conditions of glucose, maltose, or succinate treatment and in the mutant strains for genes MIG1, CYC8, and TUP1 under conditions of glucose treatment. Alternative carbon sources such as maltose and succinate affected the relative abundances of 14% of the wild-type proteins, which were mainly grouped into the carbohydrate metabolism category, with the glycolysis/gluconeogenesis and citrate cycle pathways being the most highly represented pathways. Each mutant strain showed significant proteomic profile changes, affecting approximately 2% of the total proteins identified, compared to the wild-type strain under glucose treatment conditions. Similarly to the results seen with the alternative carbon sources, the changes in the mutant strains mainly affected carbohydrate metabolism, with glycolysis/gluconeogenesis and the pentose phosphate and citrate cycle pathways being the most highly represented pathways. Our results showed convergence between carbon assimilation and catabolic repression in the strains studied. Interestingly, indications of cooperative, opposing, and overlapping processes during catabolic regulation were found. We also identified target proteins of the regulatory processes, reinforcing the likelihood of catabolic repression at the posttranscriptional level. IMPORTANCE The conditions affecting catabolic regulation in X. dendrorhous are complex and suggest the presence of an alternative mechanism of regulation. The repressors Mig1, Cyc8, and Tup1 are essential elements for the regulation of the use of glucose and other carbon sources. All play different roles but, depending on the growth conditions, can work in convergent, synergistic, and complementary ways to use carbon sources and to regulate other targets for yeast metabolism. Our results reinforced the belief that further studies in X. dendrorhous are needed to clarify a specific regulatory mechanism at the domain level of the repressors as well as its relationship with those of other metabolic repressors, i.e., the stress response, to elucidate carotenogenic regulation at the transcriptomic and proteomic levels in this yeast.

show abstract

“…It is mainly considered as two pathways: dicyclic and monocyclic pathway ( Fig. 1) [5,12]. The dicyclic pathway of astaxanthin in P. rhodozyma begin with isoprene pyrophosphate (IPP) isomerase encoding the idi gene [13,14], then go through the geranylgeranyl pyrophosphate (GGPP) synthase (crtE gene)…”

Section: Introductionmentioning

confidence: 99%

Analysis of relationship between key genes and astaxanthin biosynthesis in Phaffia rhodozyma by transcript level and gene knock-out

Chen

et al. 2019

Preprint

View full text Add to dashboard Cite

Background Phaffia rhodozyma is a potential industrial source for production of natural astaxanthin. The synthetic mechanism of astaxanthin in P. rhodozyma is complex and unclear that blocked its development. Results In this study, eight genes related to dicyclic and monocyclic pathway in three different strains of P. rhodozyma were analyzed, and the relationship between the expression and astaxanthin biosynthesis was explored. Among these genes, crtYB (R=0.75, P<0.05) and asy genes (R=0.74, P<0.05) showed the most closely correlation with astaxanthin biosynthesis. In order to further study exact relationship, crtYB and asy genes were knocked out by homologous recombination. After crtYB knock-out, astaxanthin was decreased to be under detected line. It suggested crtYB played a role in dicyclic and monocyclic pathway. Meanwhile, the asy gene was in dicyclic pathway of astaxanthin biosynthesis, and its knock-out would promote the astaxanthin biosynthesis in monocyclic pathway, resulting in a 25.04% increase in astaxanthin production. Conclusion The possible rate-limiting enzymes were asy gene and crtYB illustrated by analysis of regression. Knock-out of asy and crtYB gene was great helpful to understand the synthetic pathway of astaxanthin, and significant to the industrial application of producing astaxanthin.

show abstract

Proteomic analysis of the carotenogenic yeast Xanthophyllomyces dendrorhous

Cited by 21 publications

References 51 publications

Proteomic and metabolomic analysis of the carotenogenic yeast Xanthophyllomyces dendrorhous using different carbon sources

Proteomic and metabolomic analysis of the carotenogenic yeast Xanthophyllomyces dendrorhous using different carbon sources

Convergence between Regulation of Carbon Utilization and Catabolic Repression in Xanthophyllomyces dendrorhous

Analysis of relationship between key genes and astaxanthin biosynthesis in Phaffia rhodozyma by transcript level and gene knock-out

Contact Info

Product

Resources

About