GATA transcription factor WC2 regulates the biosynthesis of astaxanthin in yeast Xanthophyllomyces dendrorhous

Abstract

Astaxanthin is a type of carotenoid widely used as powerful antioxidant and colourant in aquaculture and the poultry industry. Production of astaxanthin by yeast Xanthophyllomyces dendrorhous has attracted increasing attention due to high cell density and low requirements of water and land compared to photoautotrophic algae. Currently, the regulatory mechanisms of astaxanthin synthesis in X. dendrorhous remain obscure. In this study, we obtained a yellow X. dendrorhous mutant by Atmospheric and Room Temperature Plasma (ARTP) mutagenesis and sequenced its genome. We then identified a putative GATA transcription factor, white collar 2 (XdWC2), from the comparative genome data and verified that disruption of the XdWC2 gene resulted in a similar carotenoid profile to that of the ARTP mutant. Furthermore, transcriptomic analysis and yeast one‐hybrid (Y1H) assay showed that XdWC2 regulated the expression of phytoene desaturase gene CrtI and astaxanthin synthase gene CrtS. The yeast two‐hybrid (Y2H) assay demonstrated that XdWC2 interacted with white collar 1 (XdWC1) forming a heterodimer WC complex (WCC) to regulate the expression of CrtI and CrtS. Increase of the transcriptional levels of XdWC2 or CrtS in the wild‐type strain did not largely modify the carotenoid profile, indicating translational and/or post‐translational regulations involved in the biosynthesis of astaxanthin. Overexpression of CrtI in both the wild‐type strain and the XdWC2‐disrupted strain apparently improved the production of monocyclic carotenoid 3‐hydroxy‐3′, 4′‐didehydro‐β, ψ‐carotene‐4‐one (HDCO) rather than β‐carotene and astaxanthin. The regulation of carotenoid biosynthesis by XdWC2 presented here provides the foundation for further understanding the global regulation of astaxanthin biosynthesis and guides the construction of astaxanthin over‐producing strains.

Omics driven onboarding of the carotenoid producing red yeastXanthophyllomyces dendrorhousCBS 6938

“…Therefore, there is a need for more strong promoters. Furthermore, previous work has shown that astaxanthin metabolism is induced by oxidative stress and light (Zhang et al 2019;Tropea et al 2013;Stachowiak et al 2013;Schroeder et al 1995;Li et al 2023;Liu et al 2006;Kim et al 2005;Vázquez et al 2001), and Huang et al has discovered a homolog of the white-collar 2 transcription factor in X. dendrorhous (Huang et al 2022), hinting that X. dendrorhous may possess stress and light-inducible promoters. Transcriptomics, and particularly differential gene expression (DGE) analysis, is one way that both strong constitutive and regulated promoters could be identified.…”

“…The light response systems in X. dendrorhous are relatively unknown, save white collar (Huang et al 2022). However, systems such as cryptochrome, BLUF-domain, rhodopsin, and photolyase are known in other fungi, including a distantly related basidiomycete Ustilago maydis (Yu & Fischer 2019;Brych et al 2016;Brych et al 2021).…”

“…So far, only 8 constitutive promoters from central carbon metabolism are available (Rodríguez-Sáiz et al 2009;. Furthermore, complex but uncharacterized regulation of the astaxanthin biosynthetic pathway limits utilization of this organism as a platform for production (Rodríguez-Sáiz et al 2010;Zhang et al 2019;Tropea et al 2013;Stachowiak 2013;Schroeder & Johnson 1995;Li et al 2023;Liu et al 2006;Huang et al 2022;Kim et al 2005). Therefore, a combined genomics and transcriptomics effort could reveal metabolic regulation and yield additional promoters for future engineering, which could be key to realizing the potential of this yeast.…”

Application of Atmospheric and Room-Temperature Plasma (ARTP) to Microbial Breeding

“…ARTP technology has significantly contributed to the improvement and screening of yeast strains. It has proven effective in producing various yeast mutants with specific traits, such as high-yield yeast [ 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 ], salt-tolerant yeast [ 77 , 78 ], and acid-tolerant yeast [ 79 ], among others ( Table 2 ). Saccharomyces cerevisiae , a well-known yeast species, holds immense importance in industries like winemaking, baking, and brewing, with a historical significance dating back to ancient times [ 80 ].…”

“…Through gene knockout and ARTP mutagenesis, they obtained high-producing tyrosine mutants and conducted genome and transcriptome analysis to reveal the underlying mechanism behind the increased tyrosine production. Similarly, ARTP mutagenesis was used to generate a yellow X. dendrorhous mutant and the GATA transcription factor XdWC2 was identified as crucial in controlling astaxanthin production [ 74 ]. This research laid the foundation for understanding the global regulation of astaxanthin biosynthesis and guiding the development of high-yield astaxanthin-producing strains.…”