Heterologous carotenoid production in Saccharomyces cerevisiae induces the pleiotropic drug resistance stress response

Abstract

To obtain insight into the genome-wide transcriptional response of heterologous carotenoid production in Saccharomyces cerevisiae, the transcriptome of two different S. cerevisiae strains overexpressing carotenogenic genes from the yeast Xanthophyllomyces dendrorhous grown in carbon-limited chemostat cultures was analysed. The strains exhibited different absolute carotenoid levels as well as different intermediate profiles. These discrepancies were further sustained by the difference of the transcriptional response exhibited by the two strains. Transcriptome analysis of the strain producing high carotenoid levels resulted in specific induction of genes involved in pleiotropic drug resistance (PDR). These genes encode ABC-type and major facilitator transporters which are reported to be involved in secretion of toxic compounds out of cells. β-Carotene was found to be secreted when sunflower oil was added to the medium of S. cerevisiae cells producing high levels of carotenoids, which was not observed when added to X. dendrorhous cells. Deletion of pdr10, one of the induced ABC transporters, decreased the transformation efficiency of a plasmid containing carotenogenic genes. The few transformants that were obtained had decreased growth rates and lower carotenoid production levels compared to a pdr5 deletion and a reference strain transformed with the same genes. Our results suggest that production of high amounts of carotenoids in S. cerevisiae leads to membrane stress, in which Pdr10 might play an important role, and a cellular response to secrete carotenoids out of the cell.

Production of Lycopene in the Non-Carotenoid-Producing Yeast Yarrowia lipolytica

“…We assume that lycopene-containing cells have lower viability, so that white or less-colored revertants are more likely to be able to form colonies on agar plates than cells containing larger amounts of lycopene. The reason for this could be a susceptibility limit for intracellular lycopene, because of its antifungal property (6,7). That non-lycopene-producing cells form bigger colonies was also demonstrated in P. pastoris (11).…”

“…Because of its 11 conjugated double bonds, it shows stronger singlet oxygen quenching ability than beta-carotene and alpha-tocopherol (5), which is supposed to prevent cancer and improve health status. Lycopene is known to have antifungal activity against the human pathogen Candida albicans (6) and causes a stress response in S. cerevisiae (7). Carotenoid production was already achieved with the non-carotenoid-producing yeasts S. cerevisiae (8), Candida utilis (3,9,10), Pichia pastoris (11), and Y. lipolytica (12)(13)(14).…”

Synergistic Effects of Branched-chain Amino Acids and Phenylalanine Addition on Major Volatile Compounds in Wine during Alcoholic Fermentation

“…In all fermentation samples, the concentrations of glucose, fructose, ethanol, glycerol, acetic acid, malic acid, citric acid, lactic acid and succinic acid were determined using the HPLC method described by Verwaal et al (2010). The system was equipped with a HPX-87H Aminex ion-exchange column (300 mm × 7.8 mm, Bio-Rad Laboratories, Hercules, CA, USA), with 5 mM sulfuric acid as the mobile phase.…”

Effect of Linoleic Acids on the Release of β-Carotene from Carotenoid-Producing Saccharomyces cerevisiae into Sunflower Oil

“…'Milking' of β-carotene has been described for the algae Dunaliella salina in a bioreactor process [Hejazi et al, 2004], but to our knowledge β-carotene has not been extracted in comparable amounts from recombinant carotenoid-producing baker's yeast [Verwaal et al, 2010]. In this study, we demonstrate that the observed β-carotene release from yeast cells into sunflower oil, formerly reported by Verwaal et al [2010], seems to be caused by the effects of linoleic acid, which is the main component of sunflower oil, towards the cells. Baker's yeast is a promising host for heterologous β-carotene production because of its GRAS (generally recognized as safe) status, easy handling and the availability of diverse genetic tools.…”

“…Its strong hydrophobic character, leading to its accumulation in cellular membranes [Gruszecki and Strzalka, 2005], hinders direct removal of β-carotene out of cultures, which would be favorable with respect to an economic process. 'Milking' of β-carotene has been described for the algae Dunaliella salina in a bioreactor process [Hejazi et al, 2004], but to our knowledge β-carotene has not been extracted in comparable amounts from recombinant carotenoid-producing baker's yeast [Verwaal et al, 2010]. In this study, we demonstrate that the observed β-carotene release from yeast cells into sunflower oil, formerly reported by Verwaal et al [2010], seems to be caused by the effects of linoleic acid, which is the main component of sunflower oil, towards the cells.…”