0000000001324136
AUTHOR
Mario Aguedo
Involvement of acyl coenzyme A oxidase isozymes in biotransformation of methyl ricinoleate into gamma-decalactone by Yarrowia lipolytica.
ABSTRACT We reported previously on the function of acyl coenzyme A (acyl-CoA) oxidase isozymes in the yeast Yarrowia lipolytica by investigating strains disrupted in one or several acyl-CoA oxidase-encoding genes ( POX1 through POX5 ) (H. Wang et al., J. Bacteriol. 181:5140–5148, 1999). Here, these mutants were studied for lactone production. Monodisrupted strains produced similar levels of lactone as the wild-type strain (50 mg/liter) except for Δ pox3 , which produced 220 mg of γ-decalactone per liter after 24 h. The Δ pox2 Δpox3 double-disrupted strain, although slightly affected in growth, produced about 150 mg of lactone per liter, indicating that Aox2p was not essential for the biotra…
Optimization of Yarrowia lipolytica’s β-oxidation pathway for γ-decalactone production
The yeast Yarrowia lipolytica growing on methyl ricinoleate produces various lactones, γ-decalactone, the worthy aroma compound, 3-hydroxy-γ-decalactone without sensorial properties and two decenolides of various interest. Unfortunately, these three latter lactones are produced at high levels by this yeast, decreasing yields and complicating the extraction of γ-decalactone. In this study, the production of γ-decalactone was increased through a genetic engineering of the strain and the accumulation of the three other lactones was lowered. Theses results show that it is possible to improve the mastering of the complex β-oxidation pathway (the metabolic pathway involved in these bioconversions…
Mechanisms underlying the toxicity of lactone aroma compounds towards the producing yeast cells
M. A G U E D O , L. B E N E Y , Y. W A C H EA N D J. - M. B E L I N. 2003. Aims: To study the fundamental mechanisms of toxicity of the fruity aroma compound c-decalactone, that lead to alterations in cell viability during its biotechnological production by yeast cells; Yarrowia lipolytica that is able to produce high amounts of this metabolite was used here as a model. Methods and Results: Lactone concentrations above 150 mg l )1 inhibited cell growth, depolarized the living cells and increased membrane fluidity. Infrared spectroscopic measurements revealed that the introduction of the lactone into model phospholipid bilayers, decreased the phase transition temperature. Moreover, the H + -…
Increased electron donor and electron acceptor characters enhance the adhesion between oil droplets and cells of Yarrowia lipolytica as evaluated by a new cytometric assay.
International audience; The adhesion of methyl ricinoleate droplets to cells of the yeast Yarrowia lipolytica was investigated. A new cytometric method, relying on the double staining of fatty globules with Nile Red and of cells with Calcofluor, enabled us to quantify methyl ricinoleate droplet adhesion to cells precultured on a hydrophilic or on a hydrophobic carbon source. In this last case, droplet adsorption was enhanced and a MATS (microbial adhesion to solvents) test revealed that this increase was due to Lewis acid-base interactions and not to an increase in the hydrophobic properties of the cell surface. These preliminary results demonstrate that the developed cytometric method is p…
Interactions between bacterial surfaces and milk proteins, impact on food emulsions stability
Bacteria possess physicochemical surface properties such as hydrophobicity, Lewis acid/base and charge which are involved in physicochemical interactions between cells and interfaces. Moreover, food matrices are complex and heterogeneous media, with a microstructure depending on interactions between the components in media (van der Waals, electrostatic or structural forces, etc.). Despite the presence of bacteria in fermented products, few works have investigated how bacteria interact with other food components. The objective of the present study was to determine the effects of the surface properties of lactic acid bacteria on the stability of model food emulsions. The bacteria were added t…
Production of 3-hydroxy-γ-decalactone, the precursor of two decenolides with flavouring properties, by the yeast Yarrowia lipolytica
3-Hydroxy-γ-decalactone is the precursor of dec-2 and dec-3-en-4-olides which are valuable aroma compounds not yet produced. To promote the accumulation of this lactone, the yeast Yarrowia lipolytica was placed in different environmental conditions aiming at altering β-oxidation fluxes. The concentration of substrate, pH, aeration and dissolved oxygen level were modified. We observed an important accumulation at low aeration (0.40 molar yields) and, to a lesser extent, at lower pH (0.15). As oxygen played a key-role, we evaluated its effect at fixed dissolved oxygen and at the pH which was the most favourable to the biotransformation (pH 4.5). At 5% and 30% dissolved oxygen, yields reached …
Medium-size droplets of methyl ricinoleate are reduced by cell-surface activity in the gamma-decalactone production by Yarrowia lipolytica.
International audience; Size of methyl ricinoleate droplets during biotransformation into gamma-decalactone by Yarrowia lipolytica was measured in both homogenized and non-homogenized media. In non-homogenized but shaken medium, droplets had an average volume surface diameter d32 of 2.5 microm whereas it was 0.7 microm in homogenized and shaken medium. But as soon as yeast cells were inoculated, both diameters became similar at about 0.7 microm and did not vary significantly until the end of the culture. The growth of Y. lipolytica in both media was very similar except for the lag phase which was lowered in homogenized medium conditions.
Intracellular pH-dependent efflux of the fluorescent probe pyranine in the yeast Yarrowia lipolytica.
International audience; 8-Hydroxypyrene-1,3,6-trisulfonic acid (pyranine) can be used as a vital intracellular pH (pH(i)) indicator. In the yeast Yarrowia lipolytica, a partial efflux of the probe was detected by using the pH-independent wavelength of 415 nm. A simplified correction of the fluorescent signals was applied, enabling to show for this species a good near-neutral pH(i) maintenance capacity in a pH 3.9 medium. Octanoic acid, which is known to have toxic effects on yeast, decreased the pH(i) and increased the 260-nm-absorbing compounds leakage. However, this acid inhibited the fluorescent probe efflux linearly with its concentration suggesting a pH(i)-dependent efflux of pyranine …
Modification génétique de la levure Yarrowia lipolytica pour maîtriser la β-oxydation
Les biotechnologies font regulierement appel a la β-oxydation pour produire des composes a haute valeur ajoutee ou depolluer des effluents industriels. Cette voie metabolique est difficile a maitriser car elle implique un nombre reduit d'enzymes pour une voie tres longue. En particulier, la production d'intermediaires reactionnels est delicate car les enzymes de la voie de reconsommation sont souvent les memes que celles impliquees dans la production. Ainsi, la production de γ-decalactone a partir de ricinoleate de methyle chez Y. lipolytica implique la β-oxydation (Endrizzi et al., 1996). Cette levure est un bon modele pour etudier le metabolisme des acides gras car elle possede cinq acyl-…
Interaction of an odorant lactone with model phospholipid bilayers and its strong fluidizing action in yeast membrane
International audience; Some odorant lactones are naturally present in fruits or in fermented products; they can also be used as food additives and can be produced by microorganisms at the industrial scale by biotechnological processes. Gamma-decalactone was previously shown to have antimicrobial properties. We determined by infrared spectroscopy measurements that this compound rapidly diffused into model phospholipid bilayers (within 2 min), modifying the general physical state of a dimyristoyl-L-alpha-phosphatidylcholine (DMPC) film. In vivo, the lactone strongly increased membrane fluidity in the model yeast Yarrowia lipolytica, as evaluated by fluorescence anisotropy measurements. This …
An air-lift biofilm reactor for the production of γ-decalactones by Yarrowia lipolytica
Decalactones are interesting flavouring compounds that can be produced from ricinoleic acid. In this study, the production of lactones in biofilms using Yarrowia lipolytica is investigated. The hydrophobia of cells increased for increased aeration rates resulting in higher adhesion when the reactor wall was hydrophobic (plastic). To increase adhesion, sheets of methyl-polymethacrylate (PMMA) were added in the reactor and the production of lactones increased with the surface of plastic added, reaching 850 mg/L of 3-hydroxy-γ-decalactone for 60 cm2. In an Airlift bioreactor made of PMMA, biofilms were present at the top of the reactor for increased aeration. In the meantime, a metabolic shift…
Increased electron donor and electron acceptor characters enhance the adhesion between oil droplets and cells of Yarrowia lipolytica as evaluated by a new cytometric assay
The adhesion of methyl ricinoleate droplets to cells of the yeast Yarrowia lipolytica was investigated. A new cytometric method, relying on the double staining of fatty globules with Nile Red and of cells with Calcofluor, enabled us to quantify methyl ricinoleate droplet adhesion to cells precultured on a hydrophilic or on a hydrophobic carbon source. In this last case, droplet adsorption was enhanced and a MATS (microbial adhesion to solvents) test revealed that this increase was due to Lewis acid-base interactions and not to an increase in the hydrophobic properties of the cell surface. These preliminary results demonstrate that the developed cytometric method is promising for various app…