scholarly journals Transcriptomic Response of Saccharomyces cerevisiae during Fermentation under Oleic Acid and Ergosterol Depletion

Fermentation ◽  
2019 ◽  
Vol 5 (3) ◽  
pp. 57 ◽  
Author(s):  
Giacomo Zara ◽  
Hennie J. J. van Vuuren ◽  
Ilaria Mannazzu ◽  
Severino Zara ◽  
Marilena Budroni
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Oleic Acid ◽  
Lipid Fraction ◽  
Membrane Lipid ◽  
Ergosterol Biosynthesis ◽  
Methionine Biosynthesis ◽  
Transcriptomic Response ◽  
Hypoxic Conditions ◽  
Lipid Supplementation ◽  
Time Point

Under anaerobic/hypoxic conditions, Saccharomyces cerevisiae relies on external lipid supplements to modulate membrane lipid fraction in response to different stresses. Here, transcriptomic responses of two S. cerevisiae wine strains were evaluated during hypoxic fermentation of a synthetic must with/without ergosterol and oleic acid supplementation. In the absence of lipids, the two strains, namely EC1118 and M25, showed different behaviour, with M25 significantly decreasing its fermentation rate from the 72 h after inoculum. At this time point, the whole genome transcriptomic analysis revealed common and strain-specific responses to the lack of lipid supplementation. Common responses included the upregulation of the genes involved in ergosterol biosynthesis, as well as the seripauperin and the heat shock protein multigene families. In addition, the upregulation of the aerobic isoforms of genes involved in mitochondrial electron transport is compatible with the previously observed accumulation of reactive oxygen species in the two strains during growth in absence of lipids. Considering the strain-specific responses, M25 downregulated the transcription of genes involved in glucose transport, methionine biosynthesis and of those encoding mannoproteins required for adaptation to low temperatures and hypoxia. The identification of these pathways, which are presumably involved in yeast resistance to stresses, will assist industrial strain selection.

A biometric study of higher alcohol production in Saccharomyces cerevisiae

1990 ◽  
Vol 36 (1) ◽  
pp. 61-64 ◽  
Author(s):  
Paolo Giudici ◽  
Patrizia Romano ◽  
Carlo Zambonelli
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Key Words ◽  
Isoamyl Alcohol ◽  
Higher Alcohols ◽  
Methionine Biosynthesis ◽  
Higher Alcohol ◽  
Strain Characteristic ◽  
Alcohol Production ◽  
Flocculation Ability ◽  
Individual Strain

A hundred strains of Saccharomyces cerevisiae were examined for the ability to produce higher alcohols. In the strains tested the production of higher alcohols was found to be an individual strain characteristic and, as such, was statistically significant. The characteristics of the strains used (flocculation ability, foaming ability, killer character, and non-H2S production) were found to be uncorrelated to isobutanol and isoamyl alcohol production, whereas the production of high levels of n-propanol was found to be related to inability to produce H2S. This, in turn, suggests a link to methionine biosynthesis. Key words: Saccharomyces cerevisiae, higher alcohols, biometry, H2S production.

Oleic acid and ergosterol supplementation mitigates oxidative stress in wine strains of Saccharomyces cerevisiae

2010 ◽  
Vol 141 (3) ◽  
pp. 229-235 ◽  
Author(s):  
Sara Landolfo ◽  
Giacomo Zara ◽  
Severino Zara ◽  
Marilena Budroni ◽  
Maurizio Ciani ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Saccharomyces Cerevisiae ◽  
Oleic Acid

scholarly journals Genomic Approach to Identification of Mutations Affecting Caspofungin Susceptibility in Saccharomyces cerevisiae

2004 ◽  
Vol 48 (10) ◽  
pp. 3871-3876 ◽  
Author(s):  
Sarit Markovich ◽  
Aya Yekutiel ◽  
Itamar Shalit ◽  
Yona Shadkchan ◽  
Nir Osherov
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Wall ◽  
Ergosterol Biosynthesis ◽  
Membrane Function ◽  
Minimum Effective Concentration ◽  
Fourfold Increase ◽  
Microdilution Method ◽  
New Genes ◽  
Broth Microdilution Method ◽  
Glucan Synthesis

ABSTRACT The antifungal agent caspofungin (CAS) specifically interferes with glucan synthesis and cell wall formation. To further study the cellular processes affected by CAS, we analyzed a Saccharomyces cerevisiae mutant collection (4,787 individual knockout mutations) to identify new genes affecting susceptibility to the drug. This collection was screened for increased CAS sensitivity (CAS-IS) or increased CAS resistance (CAS-IR). MICs were determined by the broth microdilution method. Disruption of 20 genes led to CAS-IS (four- to eightfold reductions in the MIC). Eleven of the 20 genes are involved in cell wall and membrane function, notably in the protein kinase C (PKC) integrity pathway (MID2, FKS1, SMI1, and BCK1), chitin and mannan biosynthesis (CHS3, CHS4, CHS7, and MNN10), and ergosterol biosynthesis (ERG5 and ERG6). Four of the 20 genes (TPO1, VPS65, VPS25, and CHC1) are involved in vacuole and transport functions, 3 of the 20 genes (CCR4, POP2, and NPL3) are involved in the control of transcription, and 2 of the 20 genes are of unknown function. Disruption of nine additional genes led to CAS-IR (a fourfold increase of MIC). Five of these nine genes (SLG1, ERG3, VRP1, CSG2, and CKA2) are involved in cell wall function and signal transduction, and two of the nine genes (VPS67 and SAC2) are involved in vacuole function. To assess the specificity of susceptibility to CAS, the MICs of amphotericin B, fluconazole, flucytosine, and calcofluor for the strains were tested. Seven of 20 CAS-IS strains (with disruption of FKS1, SMI1, BCK1, CHS4, ERG5, TPO1, and ILM1) and 1 of 9 CAS-IR strains (with disruption of SLG1) demonstrated selective susceptibility to CAS. To further explore the importance of PKC in CAS susceptibility, the activity of the PKC inhibitor staurosporine in combination with CAS was tested against eight Aspergillus clinical isolates by the microdilution assay. Synergistic or synergistic-to-additive activities were found against all eight isolates by use of both MIC and minimum effective concentration endpoints.

scholarly journals Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae

2021 ◽  
Vol 9 ◽  
Author(s):  
Zhi-Jiao Sun ◽  
Jia-Zhang Lian ◽  
Li Zhu ◽  
Yi-Qi Jiang ◽  
Guo-Si Li ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Biosynthetic Pathway ◽  
Metabolic Flux ◽  
Feeding Strategy ◽  
Pathway Engineering ◽  
Ergosterol Biosynthesis ◽  
Sterol Esters ◽  
Storage Pool ◽  
Ergosterol Content ◽  
Dry Cell Weight

Ergosterol, a terpenoid compound produced by fungi, is an economically important metabolite serving as the direct precursor of steroid drugs. Herein, ergsosterol biosynthetic pathway modification combined with storage capacity enhancement was proposed to synergistically improve the production of ergosterol in Saccharomyces cerevisiae. S. cerevisiae strain S1 accumulated the highest amount of ergosterol [7.8 mg/g dry cell weight (DCW)] among the wild-type yeast strains tested and was first selected as the host for subsequent metabolic engineering studies. Then, the push and pull of ergosterol biosynthesis were engineered to increase the metabolic flux, overexpression of the sterol acyltransferase gene ARE2 increased ergosterol content to 10 mg/g DCW and additional overexpression of a global regulatory factor allele (UPC2-1) increased the ergosterol content to 16.7 mg/g DCW. Furthermore, considering the hydrophobicity sterol esters and accumulation in lipid droplets, the fatty acid biosynthetic pathway was enhanced to expand the storage pool for ergosterol. Overexpression of ACC1 coding for the acetyl-CoA carboxylase increased ergosterol content from 16.7 to 20.7 mg/g DCW. To address growth inhibition resulted from premature accumulation of ergosterol, auto-inducible promoters were employed to dynamically control the expression of ARE2, UPC2-1, and ACC1. Consequently, better cell growth led to an increase of ergosterol content to 40.6 mg/g DCW, which is 4.2-fold higher than that of the starting strain. Finally, a two-stage feeding strategy was employed for high-density cell fermentation, with an ergosterol yield of 2986.7 mg/L and content of 29.5 mg/g DCW. This study provided an effective approach for the production of ergosterol and other related terpenoid molecules.

scholarly journals Synthesis of Polyhydroxyalkanoate in the Peroxisome of Saccharomyces cerevisiae by Using Intermediates of Fatty Acid β-Oxidation

2001 ◽  
Vol 67 (11) ◽  
pp. 5254-5260 ◽  
Author(s):  
Yves Poirier ◽  
Nadine Erard ◽  
Jean MacDonald-Comber Petétot
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fatty Acids ◽  
Oleic Acid ◽  
Fatty Acid ◽  
Isocitrate Lyase ◽  
Dry Weight ◽  
Pha Synthase ◽  
Recombinant Yeast ◽  
Medium Chain Length ◽  
The Media

ABSTRACT Medium-chain-length polyhydroxyalkanoates (PHAs) are polyesters having properties of biodegradable thermoplastics and elastomers that are naturally produced by a variety of pseudomonads.Saccharomyces cerevisiae was transformed with thePseudomonas aeruginosa PHAC1 synthase modified for peroxisome targeting by the addition of the carboxyl 34 amino acids from the Brassica napus isocitrate lyase. The PHAC1 gene was put under the control of the promoter of the catalase A gene. PHA synthase expression and PHA accumulation were found in recombinantS. cerevisiae growing in media containing fatty acids. PHA containing even-chain monomers from 6 to 14 carbons was found in recombinant yeast grown on oleic acid, while odd-chain monomers from 5 to 15 carbons were found in PHA from yeast grown on heptadecenoic acid. The maximum amount of PHA accumulated was 0.45% of the dry weight. Transmission electron microscopy of recombinant yeast grown on oleic acid revealed the presence of numerous PHA inclusions found within membrane-bound organelles. Together, these data show that S. cerevisiae expressing a peroxisomal PHA synthase produces PHA in the peroxisome using the 3-hydroxyacyl coenzyme A intermediates of the β-oxidation of fatty acids present in the media. S. cerevisiaecan thus be used as a powerful model system to learn how fatty acid metabolism can be modified in order to synthesize high amounts of PHA in eukaryotes, including plants.

scholarly journals Cloning of Acyl-ACP Thioesterase FatA fromArachis hypogaeaL. and Its Expression inEscherichia coli

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Gao Chen ◽  
Zhen-ying Peng ◽  
Lei Shan ◽  
Ning Xuan ◽  
Gui-ying Tang ◽  
...  
Keyword(s):  
Oleic Acid ◽  
Palmitoleic Acid ◽  
Membrane Lipid ◽  
Pcr Analysis ◽  
Fatty Acid Profiles ◽  
Reading Frame ◽  
E Coli ◽  
Real Time Quantitative Pcr ◽  
Immature Seeds ◽  
Expression Inescherichia Coli

In this study, a full-length cDNA of the acyl-ACP thioesterase,AhFatA, was cloned from developing seeds ofArachis hypogaeaL. by 3′-RACE. Sequence analysis showed that the open reading frame encodes a peptide of 372 amino acids and has 50–70% identity with FatA from other plants. Real-time quantitative PCR analysis revealed thatAhFatA was expressed in all tissues ofA. hypogaeaL., but most strongly in the immature seeds harvested at 60 days after pegging. Heterologous expression ofAhFatA inEscherichia coliaffected bacterial growth and changed the fatty acid profiles of the membrane lipid, resulting in directed accumulation towards palmitoleic acid and oleic acid. These results indicate that AhFatA is at least partially responsible for determining the high palmitoleic acid and oleic acid composition ofE. coli.

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