|
| Status |
Public on Jun 01, 2008 |
| Title |
Transcriptional profiling of carotenoid producing S. cerevisiae cells |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by array
|
| Summary |
To obtain insight in the genome-wide response of heterologous carotenoid production in Saccharomyces cerevisiae, we have analyzed the transcriptome of S. cerevisiae strains overexpressing carotenogenic genes from the yeast Xanthophyllomyces dendrorhous. For this purpose, two strains producing different levels of carotenoids were grown in carbon-limited continuous cultures and genome-wide expression was analyzed. The strain producing low carotenoid levels did not exhibit a clear genome-wide transcriptional response, suggesting that low carotenoid levels do not result in cellular stress. Transcriptome analysis of a strain producing high carotenoid levels resulted in specific induction of genes involved in pleiotropic drug resistance (PDR). These genes encode ATP-binding cassette (ABC) type transporters and major facilitator transporters which are involved in secretion of toxic compounds out of cells. Our results suggest that production of high amounts of carotenoids in S. cerevisiae lead to toxicity and that these cells are prone to secrete carotenoids out of the cell. Indeed, secretion of beta-carotene into sunflower oil was observed upon addition of this hydrophobic solvent to the growth medium. Finally, it was observed that deletion of the ABC transporter pdr10, one of the induced PDR transporters, highly decreased the transformation efficiency of an episomal vector containing carotenogenic genes. The few colored transformants that were obtained had decreased growth rates and lower carotenoid production levels compared to control strains transformed with the same carotenogenic genes. These results indicate that Pdr10 might be specifically involved in carotenoid tolerance in S. cerevisiae strains.
Keywords: dose response
|
| |
|
| Overall design |
The genome wide transcriptional response of S. cerevisiae cells that heterologously produce carotenoids might provide information concerning the impact of carotenoid production on yeast physiology and might identify bottlenecks relevant for the production of these compounds. DNA microarray experiments have been proven to be a powerful tool to study the genome wide transcriptional response of S. cerevisiae to changes of the physiological state and the environment (for example 3,. Genomics approaches on cells producing heterologous metabolites to study their impact on yeast physiology have not been reported yet for S. cerevisiae. Additionally, most transcriptome studies with S. cerevisiae have been performed with cells grown in shake flasks cultures. The main drawback of shake flask cultivation is that the environment is continuously changing, which may be of high influence on carotenoid production, and interpretation of transcriptome data . Chemostat cultivation offers advantages for studies with DNA microarrays because it enables cultivation of microorganisms under tightly defined environmental conditions. An interlaboratory comparison of transcriptome data obtained in chemostat cultures has indeed demonstrated that the accuracy and reproducibility of this approach are superior to those obtained in previous studies with shake-flask cultures .
|
| |
|
| Contributor(s) |
Verwall R, van Ooyen AJ, Daran J |
| Citation(s) |
20632327 |
| |
| Submission date |
Jul 12, 2007 |
| Last update date |
Jul 01, 2016 |
| Contact name |
Jean-Marc Daran |
| E-mail(s) |
j.g.daran@tudelft.nl
|
| Phone |
+31 15 278 2412
|
| Organization name |
Delft University of Technology
|
| Department |
Department of Biotechnology
|
| Lab |
Kluyver centre for genomics of industrial organisms
|
| Street address |
Julianalaan 67
|
| City |
Delft |
| ZIP/Postal code |
2628BC |
| Country |
Netherlands |
| |
|
| Platforms (1) |
| GPL90 |
[YG_S98] Affymetrix Yeast Genome S98 Array |
|
| Samples (14)
|
| GSM209972 |
CENPK113-7D glucose limited chemostat-1 |
| GSM209973 |
CEN-PK113-7D glucose limited chemostat-2 |
| GSM209974 |
CENPK113-7D glucose limited chemostat |
| GSM209975 |
CENPK113-7D glucose limited chemostat-4 |
| GSM209976 |
Orange 01 glucose limited chemostat -1 |
| GSM209977 |
Orange01 glucose limited chemostat -2 |
| GSM209978 |
Orange01 glucose limited chemostat -3 |
| GSM209979 |
CENPK113-7D glucose/ethanol limited chemostat -1 |
| GSM209980 |
CENPK113-7D glucose/ethanol limited chemostat -2 |
| GSM209981 |
CENPK-113-7D glucose/ethanol limited chemostat -3 |
| GSM209982 |
CENPK113-7D glucose/ethanol limited chemostat -4 |
| GSM209983 |
Orange02 glucose/ethanol chemostat -1 |
| GSM209984 |
Orange02 glucose/ethanol limited chemostat -2 |
| GSM209985 |
Orange02 glucose/ethanol limited chemostat -3 |
|
| Relations |
| BioProject |
PRJNA101529 |
Less...
More...