Use of biolog methodology for optimizing the degradation of hydrocarbons by bacterial consortia.

Biolog methodology was used for the preliminary screening of different cultural conditions in order to detect the best combination/s of factors influencing the metabolic performance of bacterial consortia active in the degradation of hydrocarbons. Two microbial consortia were tested for their activity on 2 hydrocarbons (nonadecane and eicosane) in presence of the following cultural coadjuvants: vegetal oil, beta-cyclodextrine, sodium acetate, mineral solution. Tests were conducted in Biolog MT plates, where only the redox indicator of microbial growth (tetrazolium violet) and no carbon sources are provided. The microwells were filled with various combinations of hydrocarbons, microbial inoculum and coadjuvants. Blanks were prepared with the same combinations but without hydrocarbons. The results obtained show the suitability of the methodology developed to identify the most active consortium and the conditions for its best degradation performance. The efficacy of Biolog methodology (Biolog Inc., USA) for the characterization of microbial communities on the basis of the metabolic profiles obtained on specific carbon sources in the microwells of Elisa-type plates, is widely acknowledged (Garland, 1997; Pietikäinen et al., 2000; Dauber and Wolters, 2000). Due to its aptitude to simultaneously evaluate multiple microbial responses and directly organize the results, it can be adapted to meet specific study purposes (Gamo and Shji, 1999). In the present research Biolog methodology was fitted for the preliminary screening of different cultural conditions, in order to detect the best combination/s of factors influencing the metabolic performance of bacterial consortia active in the degradation of aliphatic hydrocarbons, in view of their utilization for the bioremediation of polluted sites.

Hydrocarbon degradation by a soil microbial population with beta-cyclodextrin as surfactant to enhance bioavailability.

In general the biodegradation of nonchlorinated aliphatic and aromatic hydrocarbons is influenced by their bioavailability. Hydrocarbons are very poorly soluble in water. They are easily adsorbed to clay or humus fractions in the soil, and pass very slowly to the aqueous phase, where they are metabolised by microorganisms. Surfactants that increase their solubility and improve their bioavailability can thereby accelerate degradation. Cyclodextrins are natural compounds that form soluble complexes with hydrophobic molecules. They are widely used in medicine and harmless to microorganisms and enzymes. This paper describes their in vitro effect on the biodegradative activity of a microbial population isolated from a petroleum-polluted soil, as shown by the decrease of dodecane (C12), tetracosane (C24) anthracene and naphthalene added individually as the sole carbon source to mineral medium liquid cultures. beta-cyclodextrin accelerated the degradation of all four hydrocarbons, particularly naphthalene, and influenced the growth kinetics as shown by a higher biomass yield and better utilization of hydrocarbon as a carbon and energy source. Its low cost, biocompatibility and effective acceleration of degradation make beta-cyclodextrin an attractive option for bioremediation.

Saccharomyces cerevisiae cell fatty acid composition and release during fermentation without aeration and in absence of exogenous lipids.

Medium-chain fatty-acids (MCFA) are among the main aroma compounds of fermented beverages. High concentrations of MCFA have been found in sluggish and stuck fermentations. It has been suggested that they arrest cell growth, as they may be toxic, but the causes of sluggish and stuck fermentations are still unclear. The aim of this work was to see whether the production of MCFA is related to fatty acid synthesis in the absence of exogenous lipids and aeration, and whether their increase can be regarded as a consequence, instead of the cause, of sluggish and stuck fermentations. Two possibilities were considered: (i) MCFA are produced to replace unsaturated fatty acids (UFA) for cell membrane fluidity when the lack of oxygen makes desaturation of saturated fatty acids (SFA) impossible; or (ii) MCFA are produced following the release of medium-chain acyl-CoA from the fatty acid synthetase complex (FAS) due to the accumulation of SFA, and their hydrolysis to recycle CoA-SH. In the first hypothesis, MCFA should be active in cell metabolism and be found in cell structures; in the second, MCFA should be a discard and prevalently found outside the cell. We carried out a Saccharomyces cerevisiae fermentation experiment in a synthetic, lipid-free medium without aeration. We measured the fatty acid composition of yeast cells and the amounts of MCFA and their ethyl esters in the medium throughout the fermentation. Cell growth and the oxygen content of the medium were also monitored. We found that MCFA are not immobilized in cell structures, but mainly released into the medium. Cell growth is arrested because fatty acid biosynthesis is prevented by the lack of oxygen. The higher MCFA concentrations found in sluggish and stuck fermentations can be thus regarded as an effect, and not the cause, of this arrest. Some suggestions for the prevention of these events are proposed.

Esterase activity and release of ethyl esters of medium-chain fatty acids by Saccharomyces cerevisiae during anaerobic growth.

During anaerobic fermentation, Saccharomyces cerevisiae releases large amounts of medium-chain fatty acids (MCFAs) and related ethyl esters which are very important for aromatic quality of fermented beverages. The physiological function of ester synthesis is not yet understood. As MCFAs are toxic, their conversion to esters has been proposed to be a detoxification mechanism. Esterases possess ester synthesizing ability. Throughout an anaerobic fermentation of a lipid-free synthetic medium carried out with a S. cerevisiae strain selected for wine making, we have monitored MCFA and ethyl ester production and, at the same time, measured growth and esterasic activity of intact cells. Because no correlation was found between the concentration of each fatty acid and its ethyl ester, there is no evidence that ester synthesis reduces the toxicity of MCFAs. Esterasic activity did not show any correlation with ester synthesis, but it was related to the release of MCFAs. A model is proposed in which ester synthesis is a consequence of the arrest of lipid biosynthesis resulting from a lack of oxygen. Under these conditions, an excess of acyl coenzyme A is produced, and acyl esters are formed as secondary products of reactions aimed at recovering free coenzyme A.

Chemical analysis of cryoglobulins observed in a case of essential IgG-IgA cryoglobulinaemia.

In a case of essential mixed (IgG-IgA) cryoglobulinaemia the carbohydrate and aminoacid composition of cryoglobulins was chemically analyzed. A normal aminoacid and sialic acid composition was observed, whereas the glucide percentage was augmented with respect to the normal range for the two classes of Ig.