Inhibition of marine bacteria by extracts of macroalgae: potential use for environmentally friendly antifouling paints.

Although a total ban on the use of TBT coatings is not expected in the short term, there is a growing need for environmentally safe antifouling systems. A search for new non-toxic antifoulants has been carried out among marine macroalgae. Antifouling activity of aqueous, ethanolic and dichloromethane extracts from 30 marine algae from Brittany coast (France) was examined in vitro against 35 isolates of marine bacteria. About 20% of the extracts were found to be active. The high levels of inhibitory activities against bacteria recorded in some extracts and the absence of toxicity on the development of oyster and sea urchin larvae and to mouse fibroblast growth suggests a potential for novel active ingredients in antifouling preparations.

Separation of glyceride positional isomers by silver ion chromatography.

Separation of triglyceride and diglyceride positional isomers by silver ion high-performance liquid chromatography coupled with an evaporative light-scattering detector is described. The triglyceride isomers had a fatty acid composition of CLC and CCL, where C and L were caprylic acid and linoleic acid, respectively. Diglyceride isomers, 1,2(2,3)-diglyceride and 1,3-diglyceride, which contained caprylic acid were separated too. A solvent system based on n-hexane, 2-propanol, ethyl acetate, and acetonitrile with a flow-rate of 0.8 ml/min was developed. Calibration curves of CLC and CCL were achieved with triolein as internal standard. Using this method, the incorporation of linoleic acid onto specific a position of glycerol backbone can be monitored.

Assessment of the coloring strength of brevibacterium linens strains: spectrocolorimetry versus total carotenoid extraction/quantification.

Brevibacterium linens is a major component of red-smear cheese microflora and imparts color to the cheese rind. The present study was done to evaluate carotenoid contents in 29 strains of this bacterium and to relate the color of the strains to the carotenoids present. A spectrocolorimeter was used to determine the color, and the carotenoid contents were determined by spectrophotorimetric measurement at 450 nm. Regression analysis was carried out on the color values a*, b*, and C* to determine which color value could be used to express the contents of carotenoids in B. linens biomass. The C* color value appeared to be the best estimate for correlation.

Evaluation of nitrogenous substrates such as peptones from fish:a new method based on Gompertz modeling of microbial growth.

Fish peptones from tuna, cod, salmon, and unspecified fish were compared with a casein one by using a new method based on Gompertz modeling of microbial growth. Cumulative results obtained from six species of bacteria, yeasts, and fungi showed that, in most cases, these fish peptones are very effective. Nevertheless, this study raised some questions about the standardization of fish raw material, the enzymatic hydrolysis of fish proteins, and the composition of the culture medium used for testing the peptones.

Metabolism of ricinoleic acid into gamma-decalactone: beta-oxidation and long chain acyl intermediates of ricinoleic acid in the genus Sporidiobolus sp.

In order to study differences in gamma-decalactone production in yeast, four species of Sporidiobolus were cultivated with 5% of methyl ricinoleate as the lactone substrate. In vivo studies showed different time courses of intermediates of ricinoleic acid breakdown between the four species. In vitro studies of the beta-oxidation system were conducted with crude cell extracts of Sporidiobolus spp. and with ricinoleyl-CoA (RCoA) as substrate. The beta-oxidation was detected by measuring acyl-CoA oxidase, 3-hydroxyacyl-CoA dehydrogenase activities, and acetyl-CoA production. The time courses of the CoA esters resulting from RCoA breakdown by crude extract of Sporidiobolus spp. permit the proposal of different metabolic models in the yeast. These models explained the differences observed during in vivo studies.

Characterization of Brevibacterium linens pigmentation using spectrocolorimetry.

Spectrocolorimetry was used for the evaluation of Brevibacterium linens pigmentation. A total of 23 strains of Brevibacterium linens, from various sources, were plated onto solid agar media and their color coordinates were measured and expressed by the L*a*b* colorimetric system. All the strains were located along a straight line with a hue of 66 degrees (orange color). The effect of light exposure versus storage in the dark was also investigated using this approach. Three groups with distinct behaviors were demonstrated for the 23 B. linens strains.

In situ detoxification of the fermentation medium during gamma-decalactone production with the yeast sporidiobolus salmonicolor

gamma-Decalactone (gamma-C10) is known to be highly toxic for the microorganims used for its production. In this work, three techniques were studied in order to overcome this toxicity during a bioconversion process using ricinoleic acid as precursor of the lactone: in situ trapping in oily phases, in porous hydrophobic sorbents and in beta-cyclodextrins. Oily phases added to the media (olive, Miglyol, tributyrin, and paraffin) had a protective effect on Sp. salmonicolor, and they improved the lactone production. beta-cyclodextrins, which have a hydrophobic cavity that can trap molecules such as gamma-C10, have been used to protect the yeasts. The results showed insufficient preservation of cell viability. Some sorbents (activated carbon and polystyrene-based sorbents) were successfully tested during bioconversion. In all cases viability exceeded the reference one. Nevertheless the aroma production was 30% lower than the reference. All of these solutions led to some enhancement of the cell viability during bioconversion of methyl ricinoleate to gamma-C10. For improvement of the lactone production, the oil trapping method seemed to be the best with the experimental conditions tested.

Fatty acid accumulation in the yeast Sporidiobolus salmonicolor during batch production of gamma-decalactone.

This paper provides new information about the metabolism of various fatty acids and gamma-decalactone production by yeast. An analysis of the fatty acid composition of the yeast Sporidiobolus salmonicolor during batch production of lactone with ricinoleic acid methyl ester as a precursor showed an accumulation of the gamma-decalactone precursor inside the cells. Electron microscopy of the yeasts showed the presence of large internal inclusions leading to membrane and organelle lysis and, consequently, death of the yeast. S. salmonicolor cultivated with methyl oleate did not produce gamma-decalactone and is viable during the whole culture. Analysis of the long chain fatty acid fraction showed incorporation of methyl oleate.

Chirality of the gamma-lactones produced by Sporidiobolus salmonicolor grown in two different media.

Sporidiobolus salmonicolor is an aroma-producing yeast which gives a peach-like smell to the culture media. The enantiomeric ratios of the five gamma-lactones produced by this yeast cultivated in two different media were determined by multidimensional gas chromatography (MDGC) on a fused silica capillary column coupled to a modified beta-cyclodextrin column. These ratios remain constant during growth and are not affected by the composition of the medium. The (R)-enantiomer is highly predominant (99%) for gamma-decalactone and predominant (68-88%) for gamma-octalactone, gamma-nonalactone, and (Z6)-gamma-dodecenolactone. A ratio close to racemic was found for gamma-dodecalactone. A discussion on the metabolic origin of these lactones is based on the analysis of the enantiomeric ratios obtained. With respect to consumers' preference for products considered as "natural," microbial lactone production may represent a valuable alternative to fruit flavors. The enantiomeric lactone ratios produced by Sporidiobolus salmonicolor are compared with those reported from some fruits.

Production, Identification, and Toxicity of (gamma)-Decalactone and 4-Hydroxydecanoic Acid from Sporidiobolus spp.

During the bioconversion of ricinoleic acid to (gamma)-decalactone under controlled pH conditions, Sporidiobolus salmonicolor produced only the lactone form, while Sporidiobolus ruinenii produced both the lactone form and a precursor. By using gas chromatography-mass spectrometry and gas chromatography-Fourier transform infrared analysis techniques, the precursor was identified as 4-hydroxydecanoic acid. The levels of production in the presence of high concentrations of ricinoleic acid methyl ester differed in the two Sporidiobolus species. This difference was due on the one hand to the high sensitivity of S. salmonicolor to the lactone and on the other hand to the high level of 4-hydroxydecanoic acid produced by S. ruinenii. 4-Hydroxydecanoic acid is much less toxic to the microorganisms than the lactone. In contrast to S. ruinenii, S. salmonicolor is not able to catabolize 4-hydroxydecanoic acid.

Chirality of the gamma-lactones formed by Fusarium poae INRA 45.

A filamentous fungus, Fusarium poae INRA 45, was grown in two liquid Czapek-type media (a Czapek medium, and a yeast extract plus casaminoacids-enriched Czapek medium). Eight gamma-lactones, i.e., gamma-penta-, gamma-octa-, gamma-nona-, gamma-deca-, and gamma-dodecalactone, (6Z)-gamma- and (6E)-gamma-dodecenolactones, and a di-unsaturated gamma-dodecalactone, were tentatively, or conclusively, identified by capillary gas chromatography (GC) and coupled GC-MS, in the volatile fractions of the cultures. Kinetics of the formation of gamma-dodecalactone, (6Z)-gamma- and (6E)-gamma-dodecenolactones, (three major lactones), and gamma-dodecalactone (a minor lactone) were quantitatively studied by computerized GC integration. Variations of the (R/S) ratios of the lactone enantiomers were quantitatively studied by computerized multidimensional gas chromatography (MDGC) integration. The levels of the lactones were superior in the enriched Czapek-type medium to those of the Czapek medium, but the compositions of these media did not influence significantly the (R/S) ratios of these lactones. Formation of the enantiomers is discussed in terms of metabolism of potential precursors.