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 headspace techniques. Two strains of
Lactococcus lactis subsp.
lactis biov. diacetylactis reflecting the natural diversity of the bacterial surface properties and two ethyl esters generally involved in the cheese flavor (ethyl acetate and ethyl hexanoate) were chosen for the experiments. The results have shown that bacteria, through their surface physicochemical properties, can interact directly with aroma compounds or in an indirect way, by changing the emulsion characteristics. However, these effects depend on the physicochemical properties of both aroma compounds and bacterial surfaces.
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 be active against short-chain fatty acids, whereas Aox5 was active against molecules of all chain lengths. Mutations in Aox4 and Aox5 resulted in an increase in total Aox activity. The growth of mutant strains was analyzed. In the presence of POX1 only, strains did not grow on fatty acids, whereas POX4 alone elicited partial growth, and the growth of the double POX2-POX3-deleted mutant was normal excepted on plates containing oleic acid as the carbon source. The amounts of Aox protein detected by Western blotting paralleled the Aox activity levels, demonstrating the regulation of Aox in cells according to the POX genotype.
The yeast
Yarrowia lipolytica possesses five acyl-CoA oxidases (Aox1p to 5), the enzyme catalysing the first reaction of β-oxidation. The understanding of the specific role of each acyl-CoA oxidase ...is important to construct a yeast strain growing at a good rate and able to produce without degrading the aroma compound γ-decalactone. In this study we observed that Aox4p exhibits a slight activity on a broad spectrum of substrates and that it is involved in lactone degradation. We constructed a strain lacking this activity. Its growth was only slightly altered and it produced 10 times more lactone than the wild type in 48
h.
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 applications concerning the study of interactions between microorganisms and an emulsified hydrophobic substrates. Keywords: Adsorption; methyl ricinoleate; surface properties; Yarrowia lipolytica; cytometry
The surface of the lipid-degrading yeast, Yarrowia lipolytica, was characterized by contact angle and zeta potential (zeta) measurements. The cells were mainly hydrophilic with a negative charge that ...was only affected from pH 2 to 4. To study the effects of the surface charges on the biotransformation of methyl ricinoleate into the aroma compound, gamma-decalactone, the zeta values of the substrate droplets were modified by adding a cationic surfactant into the medium at concentrations that did not diminish cell viability: the adhesion of the lipid substrate to the cells was increased but not the overall performance of the process, therefore the adhesion is not the rate limiting here. Our methodology offers interesting perspectives for further applications.
Yarrowia lipolytica converts methyl ricinoleate to gamma-decalactone, a high-value fruity aroma compound. The highest amount of 3-hydroxy-gamma-decalactone produced by the yeast (263 mg l(-1)) ...occurred by increasing the k(L)a up to 120 h(-1) at atmospheric pressure; above it, its concentration decreased, suggesting a predominance of the activity of 3-hydroxyacyl-CoA dehydrogenase. Cultures were grown under high-pressure, i.e., under increased O(2) solubility, but, although growth was accelerated, gamma-decalactone production decreased. However, by applying 0.5 MPa during growth and biotransformation gave increased concentrations of dec-2-en-4-olide and dec-3-en-4-olide (70 mg l(-1)).
Chasmagnathus granulata phosphoenolpyruvate carboxykinase (PEPCK) cDNA from jaw muscle was cloned and sequenced, showing a specific domain to bind phosphoenolpyruvate in addition to the kinase-1 and ...kinase-2 motifs to bind guanosine triphosphate (GTP) and Mg
2+, respectively, specific for all PEPCKs. In the kinase-1 motifs the GK was changed to RK. The first 19 amino acids of the putative enzyme contain hydrophobic amino acids and hydroxylated residues specific to a mitochondrial type signal. The PEPCK is expressed in hepatopancreas, muscles, nervous system, heart, and gills. Hyperosmotic stress for 24 h increased the PEPCK mRNA level, gluconeogenic and PEPCK activities in muscle.
Surfactants play a key role in the biotechnological degradation of hydrophobic substrates, however this role is often misunderstood. During the biotransformation of methyl ricinoleate into the aroma ...compound γ-decalactone by the yeast Yarrowia lipolytica, a direct contact occurs between the surface of the cells and the small droplets of substrate. The impact of a series of surfactants on this process was investigated. Both ionic surfactants tested were toxic towards the yeast. This effect may be linked to a decrease in the cell membrane integrity. The interfacial area of the emulsion varied according to the non-ionic surfactant used, and this factor was correlated with the productivity of the biotransformation. By evaluating the effect of surfactants on the capacity of the cells to adhere to decane (MATH test), it was shown that the adhesion of methyl ricinoleate on yeast surface is not a rate-limiting point for the process.
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