Characterization of the two intracellular lipases of Y. lipolytica encoded by TGL3 and TGL4 genes: New insights into the role of intracellular lipases and lipid body organisation

Dulermo, Thierry; Tréton, Brigitte; Béopoulos, Athanasios; Gnankon, Affoué Philomène Kabran; Haddouche, Ramdane; Nicaud, Jean‐Marc

doi:10.1016/j.bbalip.2013.07.001

Cited by 91 publications

(87 citation statements)

References 48 publications

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“…The latter is carried out by the action of lipases and hydrolases [60] in response to the change of cellular or environmental conditions. Deletion of one or two intracellular lipases, YlTGL3 and YlTGL4 led to a two-fold increase in the capacity of cell to accumulate lipid [61]. Degradation of released fatty acids take places in the peroxisome via β-oxidation pathway.…”

Section: Discussionmentioning

confidence: 99%

Engineering Yarrowia lipolytica for Enhanced Production of Lipid and Citric Acid

Abghari

Chen

2017

Fermentation

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Increasing demand for plant oil for food, feed, and fuel production has led to food-fuel competition, higher plant lipid cost, and more need for agricultural land. On the other hand, the growing global production of biodiesel has increased the production of glycerol as a by-product. Efficient utilization of this by-product can reduce biodiesel production costs. We engineered Yarrowia lipolytica (Y. lipolytica) at various metabolic levels of lipid biosynthesis, degradation, and regulation for enhanced lipid and citric acid production. We used a one-step double gene knock-in and site-specific gene knock-out strategy. The resulting final strain combines the overexpression of homologous DGA1 and DGA2 in a POX-deleted background, and deletion of the SNF1 lipid regulator. This increased lipid and citric acid production in the strain under nitrogen-limiting conditions (C/N molar ratio of 60). The engineered strain constitutively accumulated lipid at a titer of more than 4.8 g/L with a lipid content of 53% of dry cell weight (DCW). The secreted citric acid reached a yield of 0.75 g/g (up to~45 g/L) from pure glycerol in 3 days of batch fermentation using a 1-L bioreactor. This yeast cell factory was capable of simultaneous lipid accumulation and citric acid secretion. It can be used in fed-batch or continuous bioprocessing for citric acid recovery from the supernatant, along with lipid extraction from the harvested biomass.

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Section: Discussionmentioning

confidence: 99%

Engineering Yarrowia lipolytica for Enhanced Production of Lipid and Citric Acid

Abghari

Chen

2017

Fermentation

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“…In Y. lipolytica, DGA2 and TGL4 are the most important genes regulating TAG synthesis and remobilization, respectively (19,24). It is therefore likely that they play a role in the lysis of lipid bodies.…”

Section: Resultsmentioning

confidence: 99%

“…Under this scenario, cell death would be caused by the liberation of FFAs into the cytoplasm. In order to better understand what happens when ⌬Ylpex11 is grown in YNBD 0.5 O 3 , the expression levels of genes involved in TAG storage (such as DGA1, LRO1, and particularly DGA2 [23]) and TAG remobilization (such as Tgl4p [24,53]) were analyzed using RT- PCR (Fig. 9).…”

Section: Resultsmentioning

confidence: 99%

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Role of Pex11p in Lipid Homeostasis in Yarrowia lipolytica

Dulermo

Gamboa-Meléndez

et al. 2015

Eukaryot Cell

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cPeroxisomes are essential organelles in the cells of most eukaryotes, from yeasts to mammals. Their role in ␤-oxidation is particularly essential in yeasts; for example, in Saccharomyces cerevisiae, fatty acid oxidation takes place solely in peroxisomes. In this species, peroxisome biogenesis occurs when lipids are present in the culture medium, and it involves the Pex11p protein family: ScPex11p, ScPex25p, ScPex27p, and ScPex34p. Yarrowia lipolytica has three Pex11p homologues, which are YALI0C04092p (YlPex11p), YALI0C04565p (YlPex11C), and YALI0D25498p (Pex11/25p). We found that these genes are regulated by oleic acid, and as has been observed in other organisms, YlPEX11 deletion generated giant peroxisomes when mutant yeast were grown in oleic acid medium. Moreover, ⌬Ylpex11 was unable to grow on fatty acid medium and showed extreme dose-dependent sensitivity to oleic acid. Indeed, when the strain was grown in minimum medium with 0.5% glucose and 3% oleic acid, lipid body lysis and cell death were observed. Cell death and lipid body lysis may be partially explained by an imbalance in the expression of the genes involved in lipid storage, namely, DGA1, DGA2, and LRO1, as well as that of TGL4, which is involved in lipid remobilization. TGL4 deletion and DGA2 overexpression resulted in decreased oleic acid sensitivity and delayed cell death of ⌬Ylpex11, which probably stemmed from the release of free fatty acids into the cytoplasm. All these results show that YlPex11p plays an important role in lipid homeostasis in Y. lipolytica.

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“…Acetyl-CoA carboxylase (ACC1) and thioesterase (TE) are examples of these targets. Disruption of other acyl-CoA synthetases (e.g., YlFat1) [81] and overexpression of intracellular lipases (e.g., TGL3 and TGL4) [126] may also contribute to a larger pool of free fatty acids. Microscopic examination of resting cells may reveal more information about the status of remaining fatty acids.…”

Section: Discussionmentioning

confidence: 99%

Combinatorial Engineering of Yarrowia lipolytica as a Promising Cell Biorefinery Platform for the de novo Production of Multi-Purpose Long Chain Dicarboxylic Acids

2017

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This proof-of-concept study establishes Yarrowia lipolytica (Y. lipolytica) as a whole cell factory for the de novo production of long chain dicarboxylic acid (LCDCA-16 and 18) using glycerol as the sole source of carbon. Modification of the fatty acid metabolism pathway enabled creating a pool of fatty acids in a β-oxidation deficient strain. We then selectively upregulated the native fatty acid ω-oxidation pathway for the enhanced terminal oxidation of the endogenous fatty acid precursors. Nitrogen-limiting conditions and leucine supplementation were employed to induce fatty acid biosynthesis in an engineered Leu -modified strain. Our genetic engineering strategy allowed a minimum production of 330 mg/L LCDCAs in shake flask. Scale up to a 1-L bioreactor increased the titer to 3.49 g/L. Our engineered yeast also produced citric acid as a major by-product at a titer of 39.2 g/L. These results provide basis for developing Y. lipolytica as a safe biorefinery platform for the sustainable production of high-value LCDCAs from non-oily feedstock.

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Characterization of the two intracellular lipases of Y. lipolytica encoded by TGL3 and TGL4 genes: New insights into the role of intracellular lipases and lipid body organisation

Cited by 91 publications

References 48 publications

Engineering Yarrowia lipolytica for Enhanced Production of Lipid and Citric Acid

Engineering Yarrowia lipolytica for Enhanced Production of Lipid and Citric Acid

Role of Pex11p in Lipid Homeostasis in Yarrowia lipolytica

Combinatorial Engineering of Yarrowia lipolytica as a Promising Cell Biorefinery Platform for the de novo Production of Multi-Purpose Long Chain Dicarboxylic Acids

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