Synergism among lactic acid, sulfite, pH and ethanol in alcoholic fermentation of Saccharomyces cerevisiae (PE-2 and M-26)

The industrial production of ethanol is affected mainly by contamination by lactic acid bacteria besides others factors that act synergistically like increased sulfite content, extremely low pH, high acidity, high alcoholic content, high temperature and osmotic pressure. In this research two strains of Saccharomyces cerevisiae PE-2 and M-26 were tested regarding the alcoholic fermentation potential in highly stressed conditions. These strains were subjected to values up to 200 mg NaHSO3 l(-1), 6 g lactic acid l(-1), 9.5% (w/v) ethanol and pH 3.6 during fermentative processes. The low pH (3.6) was the major stressing factor on yeasts during the fermentation. The M-26 strain produced higher acidity than the other, with higher production of succinic acid, an important inhibitor of lactic bacteria. Both strains of yeasts showed similar performance during the fermentation, with no significant difference in cell viability.

Effect of sugar catabolite repression in correlation with the structural complexity of the nitrogen source on yeast growth and fermentation

Biomass and ethanol production by industrial Saccharomyces cerevisiae strains were strongly affected by the structural complexity of the nitrogen source during fermentation in media containing galactose, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low galactose concentrations independent of nitrogen supplementation. At high sugar concentrations altered patterns of galactose utilisation were observed. Biomass accumulation and ethanol production depended on the nature of the nitrogen source and were different for baking and brewing ale and lager strains. Baking yeast showed improved galactose fermentation performance in the medium supplemented with casamino acids. High biomass production was observed with peptone and casamino acids for the ale brewing strain, however high ethanol production was observed only in the presence of casamino acids. Conversely, peptone was the nitrogen supplement that induced higher biomass and ethanol production for the lager brewing strain. Ammonium salts always induced poor yeast performance. The results with galactose differed from those obtained with glucose and maltose which indicated that supplementation with a nitrogen source in the peptide form (peptone) was more positive for yeast metabolism, suggesting that sugar catabolite repression has a central role in yeast performance in a medium containing nitrogen sources with differing levels of structural complexity.

Efficient flotation of yeast cells grown in batch culture

A fast flotation assay was used to select new floating yeast strains. The flotation ability did not seem to be directly correlated to total extracellular protein concentration of the culture. However, the hydrophobicity of the cell was definitely correlated to the flotation capacity. The Saccharomyces strains (FLT strains) were highly hydrophobic and showed an excellent flotation performance in batch cultures without additives (flotation agents) and with no need for a special flotation chamber or flotation column. A stable and well-organized structure was evident in the dried foam as shown by scanning electron microscopy which revealed its unique structure showing mummified cells (dehydrated) attached to each other. The attachment among the cells and the high protein concentration of the foams indicated that proteins might be involved in the foam formation. The floating strains (strains FLT) which were not flocculent and showed no tendency to aggregate, were capable of growing and producing ethanol in a synthetic medium containing high glucose concentration as a carbon source. The phenomenon responsible for flotation seems to be quite different from the flocculation phenomenon. (C) 1996 John Wiley & Sons, Inc.

Histochemical study of the extensor digitorum longus and soleus muscles in alcoholic rats

The extensor digitorum longus (EDL) and soleus (SOL) muscle fibres from albino rats submitted to experimental chronic alcoholism were evaluated in accordance with their metabolic and morphometric profiles. Twenty-seven male animals aged 4 months and weighing approximately 400 g were used. The animals were divided into three groups: control, isocaloric and alcoholic and sacrifices were carried out after 5, 10 and 15 months. The muscles were dissected, removed, cross-sectioned in a cryostat and submitted to the NADH (nicotinamide adenine dinucleotide) reaction. The SO (slow-twitch-oxidative), FG (fast-twitch-glycolytic) and FOG (fast-twitch-oxidative-glycolytic) muscle fibre types exhibited a polygonal, triangular or rounded shape and did not present noteworthy modifications in either muscles during the study. The cross-sectional areas of the fibres from the studied muscles did not present significant differences during the observations. Fibre area behaved similarly in the alcoholic animals up to the 10th month, i.e. it was decreased, as also observed in the other groups. At 15 months, however, all fibres were increased, with a predominance of FG fibres in the SOL muscle. Changes in fibre population were observed mainly in the SOL muscle of alcoholic animals: SO fibres were initially increased in number but decreased after the 10th month, and the opposite was observed for the population of FG fibres. FOG fibres increased linearly in number throughout the experiment. The statistical analysis showed nevertheless that the fibre population and cross-sectional area changes were not significant. In the alcoholic animals quantitative variations of muscle fibres were more evident in the SOL muscle, suggesting that the SOL muscle is more sensitive to the toxic action of ethanol. The results concerning the increased fibre diameter in alcoholic animals would be associated with muscle oedema induced directly or indirectly by the ethanol.

Structural complexity of the nitrogen source and influence on yeast growth and fermentation

The structural complexity of the nitrogen source strongly affects both biomass and ethanol production by industrial strains of Saccharomyces cerevisiae, during fermentation in media containing glucose or maltose, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Diauxie was observed at low glucose and maltose concentrations independent of nitrogen supplementation. At high sugar concentrations diauxie was not easily observed. and growth and ethanol production depended on the nature of the nitrogen source. This was different for baking and brewing ale and lager yeast strains. Sugar concentration had a strong effect on the shift from oxido-fermentative to oxidative metabolism. At low sugar concentrations, biomass production was similar under both peptone and casamino acid supplementation. Under casamino acid supplementation, the time for metabolic shift increased with the glucose concentration, together with a decrease in the biomass production. This drastic effect on glucose fermentation resulted in the extinction of the second growth phase, probably due to the loss of cell viability. Ammonium salts always induced poor yeast performance. In general, supplementation with a nitrogen source in the peptide form (peptone) was more positive for yeast metabolism, inducing higher biomass and ethanol production, and preserving yeast viability, in both glucose and maltose media, for baking and brewing ale and lager yeast strains. Determination of amino acid utilization showed that most free and peptide amino acids present, in peptone and casamino acids, were utilized by the yeast, suggesting that the results described in this work were not due to a nutritional status induced by nitrogen limitation.

AN IMPROVED CULTURE-MEDIUM FOR DETECTING LIVE YEAST PHASE CELLS OF PARACOCCIDIOIDES-BRASILIENSIS

The plating efficiency of standard mycological media such as brain heart infusion (BHI) agar is poor for Paracoccidioides brasiliensis. We prepared a water-extract of yeast phase cells of P. brasiliensis and examined it for growth-enhancing activity for the fungus. The water-extract, when added to BHI agar to a concentration of 5%, improved the plating efficiency of the medium for the fungus to some extent, but the degree of improvement was considerably varied among P. brasiliensis isolates. By contrast, when the water-extract was added in combination with horse serum (4%), the plating efficiency was highly improved (to 94-99%) for all the P. brasiliensis isolates employed. The growth-enhancing factor(s) in the water-extract was heat-stable and heating at 120-degrees-C for 15 min had little, if any, effect on growth-enhancing activity.

Flotation for cell recovery from small volumes of yeast cultures

Flotation or cell recovery in foams (proportion of the total cells in the medium transferred to the foam) and flotation efficiency (proportion of the cells transferred from an initial volume of medium equal to the residual volume after flotation) are functions of time, aeration rate, initial volume of medium, and initial concentration of cells. Cell recovery reached constant values (around 96.4 +/- 6.3%) and flotation efficiency decreased (owing to increases in the liquid content of the foam), with increases in air how rate (above 6-7 ml air s(-1)) and volumes of medium (above 11 ml) added to the column. Increases in concentration of cells in the medium led to increases in the concentration of cells in the foam.

On-line preconcentration and speciation analysis of Cr(III) and Cr(VI) using baker's yeast cells immobilised on controlled pore glass

A study was undertaken to evaluate Saccharonzyces cerevisiae as a substrate for the biosorption of Cr(III) and Cr(VI) aiming to the selective determination of these species in aqueous solutions. The yeast cells were covalently immobilised on controlled pore glass (CPG), packed in a minicolumn and incorporated in an on-line flow injection system. The effect of chemical and physical variables affecting the biosorption process was tested in order to select the optimal analytical conditions for the Cr retention by S. cerevisiae. Cr(III) was retained by the immobilised cells and Cr(VI) were retained by CPG. The speciation was possible by selective and sequential elution of Cr(III) with 0.05 mol L-1 HCl and 2.0 mol L-1 HNO3 for Cr(VI). The influence of some concomitant ions up to 20 mg L-1 was also tested. Quantitative determinations of Cr were carried out by means of inductively coupled plasma optical emission spectrometry (ICP OES). Preconcentration factors of 12 were achieved for Cr(III) and 5 for Cr(VI) when 1.7 mL of sample were processed reaching detection limits of 0.45 for Cr(III) and 1.5 mu g L-1 for Cr(VI). The speciation of inorganic Cr in different kinds of natural waters was performed following the proposed method. Spiked water samples were also analysed and the recoveries were in all cases between 81 and 103%. (c) 2005 Elsevier B.V. All rights reserved.

Histochemical and SEM evaluation of the neuromuscular junctions from alcoholic rats

In the present study morphological changes occurring in the neuromuscular junctions (NMJ) of the extensor digitorum longus (EDL) and soleus muscles from albino rats (Rattus norvegicus) submitted to experimental chronic alcoholism were evaluated. Seventy two male animals aged 4 months and weighing on average 400 g were divided into three groups: control, alcoholic and isocaloric. Six rats from each group were anesthetized and sacrificed after 5, 10, 15 and 18 months. The NMJ did not show detectable morphological changes in either muscle after treatment when examined by light microscopy. With respect to the dimensions, statistical analysis demonstrated a tendency to a statistically significant treatment x time interaction for the length of soleus muscle NMJ. The ultrastructural study, however, revealed that the NMJ of the soleus muscle of animals submitted to 18 months of experimental alcoholism presented important morphological alterations. Characteristically, the NMJ of these muscles is located on an elevation on the surface of the muscle fiber, presenting a regular round, oval or elliptical shape and continuous and not very deep synaptic grooves. Approximately 30% of the NMJ of alcoholic rats are irregular in shape, with the sarcolemmal elevations typical of the synapse region being flattened on at least one side, with discontinuous synaptic grooves, and deep and punctiform contacts of the synaptic buds. These data suggest that, although skeletal muscle has a greater natural resistance against the direct or indirect effects of alcohol, some submicroscopic morphological alterations are detectable in the NMJ, especially in muscles with oxidative metabolism (soleus) following long periods of ingestion of alcohol. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Yeast biodiversity from oleic ecosystems: Study of their biotechnological properties

The aim of this study was to know the yeast biodiversity from fresh olive (Olea europaea L.) fruits, olive paste (crush olives) and olive pomace (solid waste) from Arbequina and Cornicabra varieties. Yeasts were isolated from fruits randomly harvested at various olive groves in the region of Castilla La Mancha (Spain). Olive paste and pomace, a byproduct of the processing of this raw material, were also collected in sterile flasks from different oil mills. Molecular identification methodology used included comparison of polymerase chain reaction (PCR) amplicons of their 5.8S rRNA gene and internal transcribed spacers ITS1 and ITS2 followed by restriction pattern analysis (RFLP). For some species, sequence analysis of the 5.8S rDNA gene was necessary. The results were compared to sequences held in public databases (BLAST). These techniques allowed to identify fourteen different species of yeasts, belonging to seven different genera (Zygosaccharomyces, Pichia, Lachancea, Kluyveromyces, Saccharomyces, Candida, Torulaspora) from the 108 yeast isolates. Species diversity was thus considerable: Pichia caribbica, Zygosaccharomyces fermentati (Lachancea fermentati) and Pichia holstii (Nakazawaea holstii) were the most commonly isolated species, followed by Pichia mississippiensis, Lachancea sp., Kluyveromyces thermotolerans and Saccharomyces rosinii. The biotechnological properties of these isolates, was also studied. For this purpose, the activity of various enzymes (beta-glucosidase, beta-glucanase, carboxymethylcellulase, polygalacturonase, peroxidase and lipase) was evaluated. It was important that none of species showed lipase activity, a few had cellulase and polygalacturonase activities and the majority of them presented beta-glucanase, beta-glucosidase and peroxidase activities. (C) 2010 Elsevier Ltd. All rights reserved.