Lactate and Amino Acid Catabolism in the Cheese-Ripening Yeast
            <i>Yarrowia lipolytica</i>

Mansour, S.; Beckerich, Jean-Marie; Bonnarme, Pascal

doi:10.1128/aem.01519-08

Cited by 45 publications

(37 citation statements)

References 21 publications

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“…The DNA, RNA extraction and real-time PCR were performed as described elsewhere [23,24]. The primer sequences encoding the opai were: opaiF (5′-TCCCGATTACGCTGACAAGAC-3′) and opaiR (5′-CTCCATGTCATCCAGCACCAT-3′).…”

Section: Methodsmentioning

confidence: 99%

Genetic engineering of Yarrowia lipolytica for enhanced production of trans-10, cis-12 conjugated linoleic acid

Zhang

Chen

et al. 2013

Microb Cell Fact

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BackgroundConjugated linoleic acid (CLA) has been extensively studied for decades because of its health benefits including cancer prevention, anti-atherogenic and anti-obesity effects, and modulation of the immune system. We previously described the production of trans-10, cis-12 CLA in Yarrowia lipolytica by expressing the gene coding for linoleic acid isomerase from Propionibacterium acnes (pai). However the stable strain produced CLA at about 0.08% of dry cell weight (DCW), a level of production which was not high enough for practical applications. The goal of the present study was to enhance production of CLA by genetic engineering of Y. lipolytica strains.ResultsWe have now co-expressed the delta 12-desaturase gene (FADS12, d12) from Mortierella alpina together with the codon-optimized linoleic acid isomerase (opai) gene in Y. lipolytica, expressed under the control of promoter hp16d modified by fusing 12 copies of UAS1B to the original promoter hp4d. A multi-copy integration plasmid was used to further enhance the expression of both genes. Using glucose as the sole carbon source, the genetically-modified Y. lipolytica produced trans-10, cis-12-CLA at a level of up to 10% of total fatty acids and 0.4% of DCW. Furthermore, when the recombinant yeast was grown with soybean oil, trans-10, cis-12-CLA now accumulated at a level of up to 44% of total fatty acids, which represented 30% of DCW after 38.5 h of cultivation. In addition, trans-10, cis-12-CLA was also detected in the growth medium up to 0.9 g/l.ConclusionsWe have successfully produced trans-10, cis-12-CLA with a titre of 4 g/l of culture (3.1 g/l in cells and 0.9 g/l in culture medium). Our results demonstrate the potential use of Y. lipolytica as a promising microbial cell factory for trans-10, cis-12-CLA production.

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Section: Methodsmentioning

confidence: 99%

Genetic engineering of Yarrowia lipolytica for enhanced production of trans-10, cis-12 conjugated linoleic acid

Zhang

Chen

et al. 2013

Microb Cell Fact

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“…Major differences in global metabolism between Y. lipolytica and other ripening yeasts, such as D. hansenii and K. lactis, have already been highlighted. Whereas these yeasts first assimilate lactate as a carbon and energy source, followed by amino acids, Y. lipolytica preferentially degrades amino acids (14). Since amino acid degradation products have a major impact on cheese organoleptic properties, the study of sulfur metabolism in Y. lipolytica is of major interest for understanding VSC production.…”

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confidence: 99%

New Insights into Sulfur Metabolism in Yeasts as Revealed by Studies of Yarrowia lipolytica

Hébert

Forquin-Gomez

Roux

et al. 2013

Appl Environ Microbiol

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Yarrowia lipolytica, located at the frontier of hemiascomycetous yeasts and fungi, is an excellent candidate for studies of metabolism evolution. This yeast, widely recognized for its technological applications, in particular produces volatile sulfur compounds (VSCs) that fully contribute to the flavor of smear cheese. We report here a relevant global vision of sulfur metabolism in Y. lipolytica based on a comparison between high-and low-sulfur source supplies (sulfate, methionine, or cystine) by combined approaches (transcriptomics, metabolite profiling, and VSC analysis). The strongest repression of the sulfate assimilation pathway was observed in the case of high methionine supply, together with a large accumulation of sulfur intermediates. A high sulfate supply seems to provoke considerable cellular stress via sulfite production, resulting in a decrease of the availability of the glutathione pathway's sulfur intermediates. The most limited effect was observed for the cystine supply, suggesting that the intracellular cysteine level is more controlled than that of methionine and sulfate. Using a combination of metabolomic profiling and genetic experiments, we revealed taurine and hypotaurine metabolism in yeast for the first time. On the basis of a phylogenetic study, we then demonstrated that this pathway was lost by some of the hemiascomycetous yeasts during evolution.

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“…The results showed that lowering the pH from 6.0 to 4.6 decreased the growth of S. xylosus (42,43). The second reason is that competition for amino acids may occur in the presence of Y. lipolytica 1E07, which is known to preferentially consume amino acids at the expense of lactate (26). Lincoln et al (20) found that seven Staphylococcus aureus strains required arginine, proline, cysteine, valine, leucine, and glycine for growth.…”

Section: Discussionmentioning

confidence: 99%

Investigation of Associations of Yarrowia lipolytica , Staphylococcus xylosus , and Lactococcus lactis in Culture as a First Step in Microbial Interaction Analysis

Mansour

Bailly

Landaud

et al. 2009

Appl Environ Microbiol

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The interactions that may occur between microorganisms in different ecosystems have not been adequately studied yet. We investigated yeast-bacterium interactions in a synthetic medium using different culture associations involving the yeast Yarrowia lipolytica 1E07 and two bacteria, Staphylococcus xylosus C2a and Lactococcus lactis LD61. The growth and biochemical characteristics of each microorganism in the different culture associations were studied. The expression of genes related to glucose, lactate, and amino acid catabolism was analyzed by reverse transcription followed by quantitative PCR. Our results show that the growth of Y. lipolytica 1E07 is dramatically reduced by the presence of S. xylosus C2a. As a result of a low amino acid concentration in the medium, the expression of Y. lipolytica genes involved in amino acid catabolism was downregulated in the presence of S. xylosus C2a, even when L. lactis was present in the culture. Furthermore, the production of lactate by both bacteria had an impact on the lactate dehydrogenase gene expression of the yeast, which increased up to 30-fold in the three-species culture compared to the Y. lipolytica 1E07 pure culture. S. xylosus C2a growth dramatically decreased in the presence of Y. lipolytica 1E07. The growth of lactic acid bacteria was not affected by the presence of S. xylosus C2a or Y. lipolytica 1E07, although the study of gene expression showed significant variations.

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Lactate and Amino Acid Catabolism in the Cheese-Ripening Yeast Yarrowia lipolytica

Cited by 45 publications

References 21 publications

Genetic engineering of Yarrowia lipolytica for enhanced production of trans-10, cis-12 conjugated linoleic acid

Genetic engineering of Yarrowia lipolytica for enhanced production of trans-10, cis-12 conjugated linoleic acid

New Insights into Sulfur Metabolism in Yeasts as Revealed by Studies of Yarrowia lipolytica

Investigation of Associations of Yarrowia lipolytica , Staphylococcus xylosus , and Lactococcus lactis in Culture as a First Step in Microbial Interaction Analysis

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