Unknown,Transcriptomics,Genomics,Proteomics

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Dynamic transcriptional response of the reference Saccharomyces cerevisiae strain and homozygous CCC2 deletion mutant strain to copper

ABSTRACT: Dynamic transcriptional response of S. cerevisiae cells to copper impulse was investigated in both HO deletion strain used as the reference strain and the mutant strain lacking CCC2 gene which were grown in continuous cultures using a copper-deficient defined medium. Copper was introduced into the medium as an impulse so as to reach a copper sulfate concentration of 0.5 mM. Samples were collected within the first two hours following the copper addition (1st, 5th, 10th, 15th, 20th, 25th, 30th, 60th, 120th minutes), in addition to the steady-state sampling.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Sebnem Oc

PROVIDER: E-MTAB-9155 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Saccharomyces cerevisiae has become a popular host for production of non-native compounds. The metabolic pathways involved generally require a net input of energy. To maximize the ATP yield on sugar in S. cerevisiae, industrial cultivation is typically performed in aerobic, sugar-limited fed-batch reactors which, due to constraints in oxygen transfer and cooling capacities, have to be operated at low specific growth rates. Because intracellular levels of key metabolites and cellular energy status are growth-rate dependent, slow growth can significantly affect biomass-specific productivity. Using an engineered Saccharomyces cerevisiae strain expressing a heterologous pathway for resveratrol production as a model energy-requiring product, the impact of specific growth rate on yeast physiology and productivity was investigated in aerobic, glucose-limited chemostat cultures. Stoichiometric analysis revealed that de novo resveratrol production from glucose requires a net input of 2 moles of ATP per mole of produced resveratrol. The biomass-specific production rate of resveratrol showed a strong positive correlation with the specific growth rate. At low growth rates, a substantial fraction of the carbon source was invested in cellular maintenance-energy requirements (e.g., 27% at 0.03 h-1). This distribution of resources was unaffected by resveratrol production. Formation of the by-products coumaric, phloretic and cinnamic acid had no detectable effect on maintenance energy requirement and yeast physiology in the chemostats. Expression of the heterologous pathway led to marked differences in transcript levels in the resveratrol-producing strain, including increased expression levels of genes involved in pathways for precursor supply (e.g., ARO7 and ARO9 involved in phenylalanine biosynthesis). The observed strong differential expression of many glucose-responsive genes in the resveratrol producer as compared to a congenic reference strain could be explained from higher residual glucose concentrations and higher relative growth rates in cultures of the resveratrol producer. De novo resveratrol production by engineered S. cerevisiae is an energy demanding process. Resveratrol production by an engineered strain exhibited a strong correlation with specific growth rate. Since industrial production in fed-batch reactors typically involves low specific growth rates, this study emphasizes the need for uncoupling growth and product formation via energy-requiring pathways. The goal of the present study is to investigate the impact of specific growth rate on biomass-specific productivity, product yield, by-product formation and host strain physiology of an S. cerevisiae strain that was previously engineered for de novo production of resveratrol from glucose. To this end, (by)product formation, physiology and transcriptome were analysed in steady-state, glucose-limited chemostat cultures grown at different dilution rates.

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Dynamic transcriptional response of the reference Saccharomyces cerevisiae strain and homozygous CCC2 deletion mutant strain to copper

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