0000000000541708
AUTHOR
Yves Waché
Evaluation of acyl coenzyme A oxidase (Aox) isozyme function in the n- alkane-assimilating yeast Yarrowia lipolytica
ABSTRACT We have identified five acyl coenzyme A (CoA) oxidase isozymes (Aox1 through Aox5) in the n -alkane-assimilating yeast Yarrowia lipolytica , encoded by the POX1 through POX5 genes. The physiological function of these oxidases has been investigated by gene disruption. Single, double, triple, and quadruple disruptants were constructed. Global Aox activity was determined as a function of time after induction and of substrate chain length. Single null mutations did not affect growth but affected the chain length preference of acyl-CoA oxidase activity, as evidenced by a chain length specificity for Aox2 and Aox3. Aox2 was shown to be a long-chain acyl-CoA oxidase and Aox3 was found to …
Effect of reducing agents on the acidification capacity and the proton motive force of Lactococcus lactis ssp. cremoris resting cells.
International audience; Reducing agents are potential inhibitors of the microbial growth. We have shown recently that dithiothreitol (DTT), NaBH(4) and H(2) can modify the proton motive force of resting cells of Escherichia coli by increasing the membrane protons permeability [Eur. J. Biochem. 262 (1999) 595]. In the present work, the effect of reducing agents on the resting cells of Lactococcus lactis ssp. cremoris, a species widely employed in dairy processes was investigated. DTT did not affect the acidification nor the DeltapH, in contrast to the effect previously reported on E. coli. The DeltaPsi was slightly increased (30 mV) at low pH (pH 4) in the presence of 31 mM DTT or 2.6 mM NaB…
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…
Cloning and expression of genes involved in conidiation and surface properties of Penicillium camemberti grown in liquid and solid cultures.
International audience; Based on bioinformatic data on model fungi, the rodA and wetA genes encoding, respectively, a RodA hydrophobin protein and the WetA protein involved in conidiation mechanisms, were PCR-cloned and characterized for the first time in Penicillium camemberti. These results, completed by a sequence of the brlA gene (available in GenBank), which encodes a major transcriptional regulator also involved in the conidiation mechanism, were used to compare, by qRT-PCR, the expression of the three genes in liquid and solid cultures in a synthetic medium. While expression of the brlA and wetA genes increased dramatically in both culture conditions after 4 days of growth, expressio…
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…
Effect of acyl-CoA oxidase activity on the accumulation of gamma-decalactone by the yeast Yarrowia lipolytica: a factorial approach.
International audience; beta-Oxidation is a cyclic pathway involved in the degradation of lipids. In yeast, it occurs in peroxisomes and the first step is catalyzed by an acyl-CoA oxidase (Aoxp). The yeast Yarrowia lipolytica possesses several genes (POX) coding for Aoxps. This study is based on the factorial analysis of results obtained with the many POX derivative strains that have been constructed previously. The effect of interactions between Aoxps on the acyl-CoA oxidase (Aox) activity was important even at the second order. We then investigated the effect of Aox activity on growth and lactone production. Aox activity was correlated with acidification of the medium by cells and with ce…
Importance and efficiency of in-depth antimicrobial activity for the control of listeria development with nisin-incorporated sodium caseinate films
Abstract The effect of antimicrobial sodium caseinate-based films was investigated on inhibition of Listeria innocua in cheese. Nisin was incorporated into sorbitol-plasticized sodium caseinate films at 1000 IU/cm2 and the films were prepared by casting methods. Mini red Babybel® cheese was chosen as a semi-soft cheese model. The antimicrobial activity was studied based on the contact between antimicrobial films and surface-contaminated as well as in-depth contaminated cheese samples during a 7-day storage at 4 °C. The active films produced resulted in a reduction of 1.1 log CFU/g reduction in L. innocua counts in surface-inoculated cheese samples after 1 week of storage as compared to cont…
Production of Dicarboxylic Acids and Flagrances by Yarrowia lipolytica
Yeasts are excellent biocatalysts in the field of alkane and fatty acids transformation into dicarboxylic acids and lactones. Their ability to produce some diacids through simple, less expensive and more environment friendly routes than chemical pathways and to produce particular diacids (e.g. unsaturated ones) but also to transform natural substrates into lactones with a natural label has made them the subject of many researches. Although Candida species were often first studied, the development of genetic tools, the knowledge of the genome and some genomic and biotechnological particularities of Yarrowia lipolytica resulted to interesting developments with this species. This chapter aims …
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 + -…
Protection of living yeast cells by micro-organized shells of natural polyelectrolytes
International audience; Saccharomyces cerevisiae, a eukaryotic model organism, plays a key role in the oxidative stability of fermented products. In order to protect cells against environmental stresses, we report a method of modifying the cell surface architecture while maintaining the internal working properties of the system. The objective was to encapsulate living yeast cells in micro-organized polyelectrolyte shells using layer-by-layer (LbL) assembly. For the first time, the natural polyelectrolytes, β-lactoglobulin and sodium alginate, were alternately deposited on the surface of S. cerevisiae. Transmission electron microscopy coupled with immune-cytochemistry and scanning electron m…
Molecule structural factors influencing the loading of flavoring compounds in a natural-preformed capsule: Yeast cells
International audience; Yeast cells are efficient microcapsules for the encapsulation of flavoring compounds. However, as they are preformed capsules, they have to be loaded with the active. Encapsulation efficiency is to a certain level correlated with LogP. In this study, the effect of structural factors on the encapsulation of amphiphilic flavors was investigated. Homological series of carboxylic acids, ethyl esters, lactones, alcohols and ketones were encapsulated into the yeast Yarrowia lipolytica. Although, in a single homological series, the length of the molecule and thus the LogP were correlated with encapsulation efficiency (EY%), big differences were observable between series. Fo…
Isomerization and increase in the antioxidant properties of lycopene from Momordica cochinchinensis (gac) by moderate heat treatment with UV-Vis spectra as a marker.
International audience; Momordica cochinchinensis (gac) is a plant rich in lycopene. This pigment tends to solubilize in oil and get damaged during extraction. The impact of heating on cis-isomerization of oil-free lycopene in hexane was studied at 50 and 80°C during 240min with UV-Vis spectrometry, DAD-HPLC and TEAC test. The initial all-trans-form isomerized to the 13-cis isomer more rapidly at 80°C. After this treatment, 16% of the lycopene compounds were in the 9-cis-form. This isomer triggered an increase in the antioxidant properties which was detectable from concentrations above 9% and resulted in a change from 2.4 to 3.7μmol Trolox equivalent. It is thus possible to increase the bio…
Can biological structures be natural and sustainable capsules?
Flavor and fragrance molecules are used in many industrial fields such as food, cosmetics, tissues, pharmacy, agriculture (pheromones) etc. As most actives have a specific target and are fragile molecules, encapsulation processes have been developed for their use. These technologies are efficient to avoid loss of actives, dissemination out of the target and subsequent pollution, and to protect molecules up to their target. Several processes have been developed responding to the numerous situations encountered (e.g., protection against air, temperature, light, pH; masking or revealing sensorial properties of the molecule; release during the process, in the plate, in the mouth, etc.). However…
Cell surface properties affect colonisation of raw milk by lactic acid bacteria at the microstructure level
Cell localisation in the matrix microstructure of fermented food is important for microbial activity and ripening. To evaluate the importance of physicochemical factors in these phenomena, the physicochemical behaviour of four strains of lactic acid bacteria were investigated after inoculation of raw milk or milk fractions. To discriminate between physicochemical and metabolic properties, media were (i) rapidly acidified to avoid cell division or (ii) bacteria were allowed to ferment milk. To evaluate the impact of milk components, cells were dispersed in protein or fatty globule fractions. From two Lactococcus lactis subsp. lactis biovar. diacetylactis strains selected from their different…
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…
Explorer la biodiversité pour trouver de nouvelles souches microbiennes pour les innovations de produits alimentaires
Référence : 201511122729; Entre les innovations alimentaires renforçant les propriétés sensorielles ou de santé ou améliorant le procédé, la demande de souches microbiennes nouvelles est forte. Toutefois, trouver de nouvelles souches libres de droits et efficaces n’est pas une tâche facile. Dans cet article, seront présentées des voies d’innovations pour augmenter le bénéfice sensoriel de fermentations et les activités du laboratoire international «Tropical Bioresources & Biotechnology» pour découvrir de nouvelles souches et de nouveaux procédés basés sur les fermentations traditionnelles de produits tropicaux.
Increase in stability and change in supramolecular structure of β-carotene through encapsulation into polylactic acid nanoparticles
International audience; β-Carotene (BC) exhibits controversial antioxidant properties as it may act also as a prooxidant. Its stability toward oxidation depends on its dispersion form and can be increased through encapsulation. In this study, oxidation of BC from synthetic and natural origins was investigated after dispersion in Tween micelles or poly lactic acid (PLA) particles. Two oxidation systems were used: autooxidation and oxidation by xanthine oxidase-generated-reactive oxygen species. Results showed that synthetic BC formed nanometric negatively-charged particles in both Tween micelle and PLA systems, whereas the natural BC sample used was shown to be already pre-oxidised, forming …
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 …
Fermentation des protéines végétales : atouts organoleptique et nutritionnel
Référence : 201511122426; Du fait de l’augmentation de la population de la planète, la production de protéines pour l’alimentation des populations et l’évaluation de la qualité de ces protéines pour satisfaire les besoins de l’Homme pourraient devenir des questions majeures dans les décennies à venir pour l’ensemble du monde. Le développement de nouveaux aliments optimisant l’apport protéique d’origine végétale parait également important dans certaines situations (personnes âgées, sportifs…) où les besoins nutritionnels sont modifiés au plan quantitatif et qualitatif. De nombreux problèmes ont été identifiés suite à l’incorporation de protéines végétales dans nos aliments. En particulier, d…
Rôle et perspectives des études microbiologiques pour la sécurité alimentaire
International audience
Physico-chemical state influences in vitro release profile of curcumin from pectin beads
International audience; Curcumin is a polyphenolic compound with diverse effects interesting to develop health benefit products but its formulation in functional foods or in food supplement is hampered by its poor water solubility and susceptibility to alkaline conditions, light, oxidation and heat. Encapsulation of curcumin could be a mean to overcome these difficulties. In this paper, curcumin was encapsulated by ionotropic gelation method in low methoxyl pectin beads associated with different surfactants: Solutol®, Transcutol® and sodium caseinate. After encapsulation, physico-chemical properties of encapsulated curcumin such as its solubility, physical state, tautomeric forms and encaps…
Potential of nisin-incorporated sodium caseinate films to control Listeria in artificially contaminated cheese
International audience; A sodium caseinate film containing nisin (1000 IU/cm(2)) was produced and used to control Listeria innocua in an artificially contaminated cheese. Mini red Babybel cheese was chosen as a model semi-soft cheese. L. innocua was both surface- and in-depth inoculated to investigate the effectiveness of the antimicrobial film as a function of the distance from the surface in contact with the film. The presence of the active film resulted in a 1.1 log CFU/g reduction in L. innocua counts in surface-inoculated cheese samples after one week of storage at 4 degrees C as compared to control samples. With regard to in-depth inoculated cheese samples, antimicrobial efficiency wa…
Advanced fluorescence technologies help to resolve long-standing questions about microbial vitality
International audience; Advances in fundamental physical and optical principles applied to novel fluorescence methods are currently resulting in rapid progress in cell biology and physiology. Instrumentation devised in pioneering laboratories is becoming commercially available, and study findings are now becoming accessible. The first results have concerned mainly higher eukaryotic cells but many more developments can be expected, especially in microbiology. Until now, some important problems of cell physiology have been difficult to investigate due to interactions between probes and cells, excretion of probes from cells and the inability to make in situ observations deep within the cell, w…
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 …
Antioxidants and Bioactive Compounds in Food: Critical Review of Issues and Prospects
International audience; This review paper gives an insight into the effective delivery mechanisms for health-promoting substances and highlights the challenges of using antioxidants and bioactives in foods. The selection criteria for choosing bioactives and their extraction in bioavailable form with their adequate incorporation techniques and delivery mechanisms are covered. Moreover, an overview of existing methods for determination of bioactivity is given. The importance of scientifically evaluating the effects of foods or food components on consumer health before making claims about the healthiness is aligned. Finally, a scientific perspective on how to respond to the booming demand for …
Fluorescent probes to evaluate the physiological state and activity of microbial biocatalysts: A guide for prokaryotic and eukaryotic investigation
International audience; Many fluorescent techniques are employed to evaluate the viability and activity of microbial cells used in biotechnology. These techniques are sometimes complex and the interpretation of results opened to misunderstanding. Moreover, new developments are constantly proposed especially concerning a more accurate evaluation of the state of the cells including eukaryotic microorganisms. This paper aims at presenting to biotechnologists unfamiliar with fluorescence the principles of these methods and the related possible pitfalls. It focuses on probes of the physical (integrity and fluidity) and energetical (intracellular pH and membrane potential) state of the cell membr…
Asia: Introduction
International audience
Modernizing traditional fermented foods for a more sustainable and diverse food system
Traditional fermented foods are products belonging to the World Heritage that are often obtained in a more sustainable way than other foods. Over the past decades, traditional food systems evolved to globalized networks of regulated trade. However, for safety, sensorial, and nutritional issues, the way they are produced has to be modernized with a better control of the fermentation step (eg starter cultures vs spontaneous fermentation). Innovations should follow economic, and eco-friendly approaches to improve their safety and quality, creating a good balance among history, culture, and advanced biotechnologies.The production of some traditional fermented foods is well defined through Prote…
Chương IV - VẤN ĐỀ SỬ DỤNG KHÁNG SINH TRONG CHĂN NUÔI VÀ TƯƠNG LAI CỦA NGHIÊN CỨU VI SINH VẬT ĐỐI VỚI AN TOÀN THỰC PHẨM
Trung tâm Quy trình Thực phẩm và Vi sinh vật, AgroSup Dijon, Đại học Bourgogne-Franche Comté 4�1� GIỚI THIỆU Chăn nuôi là một ngành nông nghiệp được công nghiệp hóa rất nhiều trong những thập kỷ vừa qua. Những trang trại chăn nuôi có quy mô lớn dần thay đổi bức tranh hệ thống sản xuất nông nghiệp của nhiều nước trên thế giới, trong đó có Việt Nam. Lợi thế của sản xuất quy mô là tăng năng suất và giảm chi phí đơn vị trên đầu gia súc, hiệu quả cạnh tranh cao, đáp ứng nhu cầu của thị trường về số lượng. Thế nhưng điểm yếu là mức độ an toàn, do sử dụng thức ăn công nghiệp và do rủi ro dịch bệnh lây nhiễm. Một trong những vấn đề lớn là sử dụng kháng sinh trong chăn nuôi. Tại châu Âu, việc dùng k…
Traditional fermented sausage ‘Nem chua’ as a source of yeast biocatalysts efficient for the production of the aroma compound γ-decalactone
International audience; The yeast ecosystem of Nem chua, a Vietnamese traditional fermented sausage naturally rich in medium‐chain‐length lipid‐derived flavouring compounds, was investigated to select biocatalysts able to produce the C10‐fatty acid‐derived aroma compound γ‐decalactone. The total number of yeast was about 5 × 104 to 4 × 105 CFU g−1, and eighty four different species were identified from morphological, physiological and 26S rDNA characteristics, with Candida sake and Candida haemulonii being found in all samples. Six strains able to produce γ‐decalactone from castor oil were selected, of which three Yarrowia lipolytica strains were able to produce between 1 and 2 g L−1 in our…
Changes in the proton-motive force in Escherichia coli in response to external oxidoreduction potential.
International audience; The pH homeostasis and proton-motive force (Deltap) of Escherichia coli are dependent on the surrounding oxidoreduction potential (ORP). Only the internal pH value and, thus, the membrane pH gradient (DeltapH) component of the Deltap is modified, while the membrane potential (DeltaPsi) does not change in a significant way. Under reducing conditions (Eh < 50 mV at pH 7.0), E. coli decreases its Deltap especially in acidic media (21% decrease at pH 7.0 and 48% at pH 5.0 for a 850-mV ORP decrease). Measurements of ATPase activity and membrane proton conductance (CH+m) depending on ORP and pH have shown that the internal pH decrease is due to an increase in membrane prot…
Extracellular oxidoreduction potential modifies carbon and electron flow in Escherichia coli.
ABSTRACT Wild-type Escherichia coli K-12 ferments glucose to a mixture of ethanol and acetic, lactic, formic, and succinic acids. In anoxic chemostat culture at four dilution rates and two different oxidoreduction potentials (ORP), this strain generated a spectrum of products which depended on ORP. Whatever the dilution rate tested, in low reducing conditions (−100 mV), the production of formate, acetate, ethanol, and lactate was in molar proportions of approximately 2.5:1:1:0.3, and in high reducing conditions (−320 mV), the production was in molar proportions of 2:0.6:1:2. The modification of metabolic fluxes was due to an ORP effect on the synthesis or stability of some fermentation enzy…
Influence of lactose-citrate co-metabolism on the differences of growth and energetics in Leuconostoc lactis, Leuconostoc mesenteroides ssp. mesenteroides and Leuconostoc mesenteroides ssp. cremoris
International audience; The biodiversity of growth and energetics in Leuconostoc sp. has been studied in MRS lactose medium with and without citrate. On lactose alone, Ln. lactis has a growth rate double that of Ln. cremoris and Ln. mesenteroides. The pH is a more critical parameter for Ln. mesenteroides than for Ln. lactis or Ln. cremoris; without pH control Ln. mesenteroides is unable to acidify the medium under pH 4.5, while with pH control and as a consequence of a high Y(ATP) its growth is greater than Ln. lactis and Ln. cremoris. In general, lactose-citrate co-metabolism increases the growth rate, the biomass synthesis, the lactose utilisation ratio, and the production of lactate and …
Cloning and characterization of the peroxisomal acyl CoA oxidaseACO3 gene from the alkane-utilizing yeastYarrowia lipolytica
The ACO3 gene, which encodes one of the acyl-CoA oxidase isoenzymes, was isolated from the alkane-utilizing yeast Yarrowia lipolytica as a 10 kb genomic fragment. It was sequenced and found to encode a 701-amino acid protein very similar to other ACOs, 67.5% identical to Y. lipolytica Aco1p and about 40% identical to S. cerevisiae Pox1p. Haploid strains with a disrupted allele were able to grow on fatty acids. The levels of acyl-CoA oxidase activity in the ACO3 deleted strain, in an ACO1 deleted strain and in the wild-type strain, suggested that ACO3 encodes a short chain acyl-CoA oxidase isoenzyme. This narrow substrate spectrum was confirmed by expression of Aco3p in E. coli.
La fermentation en milieu solide : une innovation de rupture à (re)découvrir
Référence : 201511123537; National audience; Le principal challenge de l’industrie des biotechnologies est lié au développement d’un procédé qui est à la fois économique et efficace en utilisant des matières premières renouvelables pour délivrer des produits à haute valeur ajoutée. La fermentation en milieu solide (FMS) qui met en œuvre des microorganismes sur des matières premières biologiques est un procédé pouvant répondre à ces problématiques.
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 …
Stability of carotenoid extracts of gấc (Momordica cochinchinensis) towards cooxidation — Protective effect of lycopene on β-carotene
International audience; Momordica cochinchinensis, so-called gấc in Vietnam, is considered as a fruit with high nutritional potential. Its antioxidant property, due to a high concentration of carotenes (β-carotene and lycopene), is particularly estimated. In this study, we have investigated the degradation of carotene extracts obtained from gấc aril. These extracts were dispersed in the aqueous phase in Tween 80 micelles or were encapsulated into polylactic acid (PLA) particles. In both cases, carotenes were far less degraded than synthetic β-carotene. However, the degradation of lycopene was still rapid (around 1 mM lycopene degraded per hour), whereas β-carotene was almost not bleached. M…
Combined action of redox potential and pH on heat resistance and growth recovery of sublethally heat-damaged Escherichia coli
International audience; The combined effect of redox potential (RP) (from -200 to 500 mV) and pH (from 5.0 to 7.0) on the heat resistance and growth recovery after heat treatment of Escherichia coli was tested. The effect of RP on heat resistance was very different depending on the pH. At pH 6.0, there was no significant difference, whereas at pH 5.0 and 7.0 maximum resistance was found in oxidizing conditions while it fell in reducing ones. In sub-lethally heat-damaged cells, low reducing and acid conditions allowed growth ability to be rapidly regained, but a decrease in the redox potential and pH brought about a longer lag phase and a slower exponential growth rate, and even led to growt…
Preferential localization of Lactococcus lactis cells entrapped in a caseinate/alginate phase separated system.
International audience; This study aimed to entrap bioprotective lactic acid bacteria in a sodium caseinate/sodium alginate aqueous two-phase system. Phase diagram at pH = 7 showed that sodium alginate and sodium caseinate were not miscible when their concentrations exceeded 1% (w/w) and 6% (w/w), respectively. The stability of the caseinate/alginate two-phase system was also checked at pH values of 6.0 and 5.5. Lactococcus lactis subsp. lactis LAB3 cells were added in a 4% (w/w) caseinate/1.5% (w/w) alginate two-phase system at pH = 7. Fluorescence microscopy allowed to observe that the caseinate-rich phase formed droplets dispersed in a continuous alginate-rich phase. The distribution of …
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…
Effects of oxidoreduction potential combined with acetic acid, NaCl and temperature on the growth, acidification, and membrane properties of Lactobacillus plantarum.
International audience; The effects of oxidoreduction potential (Eh) combined with acetic acid, NaCl and temperature on the growth, acidification, and membrane properties of Lactobacillus plantarum were studied. The culture medium was set at pH 5, and two different Eh values were adjusted using nitrogen (Eh = +350 mV) or hydrogen (Eh = -300 mV) gas. In reducing condition, the growth was slowed and the acidification delayed at 37 degrees C, but not at 10 degrees C. A synergistic inhibitory effect of reducing Eh, acetic acid and NaCl was observed, mainly for delaying the lag phase before acidification. These results may be explained by changes in ATPase activity, membrane fluidity and surface…
Retention of aroma compounds by lactic acid bacteria in model food media
The interactions between aroma compounds and other particles in foods, particularly with macromolecules, have been greatly studied in order to better understand the binding of flavors in food matrices. Bacteria possess many macromolecules on their cellular surface that provide them surface properties which are involved in the physicochemical interactions between cells and interfaces. However, the interactions between bacteria and aroma compounds have not received so much attention despite the presence of bacteria in many fermented products. In order to study the retention of aroma compounds by bacteria, we have investigated the retention of esters by lactic acid bacteria with static headspa…
Improved co-oxidation of β-carotene to β-ionone using xanthine oxidase-generated reactive oxygen species in a multiphasic system
beta-Ionone, an aroma compound exhibiting flower notes, can be obtained from beta-carotene in a cooxidation system utilizing xanthine oxidase-generated reactive oxygen species (ROS). ROS have to be controlled as, although they can give rise to beta-ionone, they may also degrade it. In this work, the biotransformation of beta-carotene into beta-ionone was investigated in systems containing variable proportions of decane to extract beta-ionone before degradation. The use of 50% or 90% decane resulted in increased production yields. Tween 80, which was added to further improve the production, slightly decreased the reactivity of the medium and the extraction of beta-carotene, but increased the…