Influence of temperature and pH on S. bayanus var. uvarum growth; impact of a wine yeast interspecific hybridization on these parameters

The species Saccharomyces bayanus var. uvarum possesses interesting enological characteristics but produces high concentration of volatile fermentative compounds not desirable in Sauvignon blanc wines. Interspecific hybrids between Saccharomyces cerevisiae and S. bayanus var. uvarum were made in order to join the main parental advantages. Two hybrids were selected on the basis of their fermentation characteristics and their karyotypes, i.e. they have a different mitochondrial DNA. In order to produce these hybrids as active dry yeast to be used as starter in winemaking, their optimal environmental conditions for growth, i.e. temperature and pH, were determined as the objective of our work. Using a two-level factorial design it was found that the two parental strains have different optimal temperature but for the two strains, pH does not have a significant influence on growth. The influence of temperature on biomass productivity for hybrid strains were strictly identical, so we suppose that the main genes coding for temperature sensitivity were not contained in mitochondrial DNA, but in nuclear DNA. Moreover the reactions of hybrid strains to the temperature variations were similar to the one of S. bayanus var.uvarum. This latter strain could have a majority of genes responsible of temperature sensitivity dominant in comparison with those of the strain S. cerevisiae

Effect of ammonium concentration on alcoholic fermentation kinetics by wine yeasts for high sugar content

Kinetics of alcoholic fermentation by Saccharomyces cerevisiae wine strains in a synthetic medium with high sugar content were established for different nitrogen initial content and are presented for 4 strains. The composition of the medium was close to grape must except that the nitrogen source consisted mainly in ammonium and was varied from 120 to 290 mg N/L assimilable nitrogen. The overall nitrogen consumed was also estimated in order to determine nitrogen requirement variability. The effect of assimilable nitrogen was in general greater on sugar consumption rates than on growth and 3 kinds of effect on sugar consumption rates were observed: i) existence of an optimal initial nitrogen level for a maximal sugar consumption rate (inhibition if excess), ii) no effect of nitrogen beyond the intermediary level (saturation), iii) sugar consumption rate proportional to the initial nitrogen level (activation). In all cases, the amount of consumed nitrogen increased with its initial concentration and so did the fructophilic capacity of the strains. The optimal requirement varied from 0.62 to 0.91 mg N per g of sugars according to the different strains. There was no general correlation between the sugar assimilation rates and the nitrogen requirement

The role of malic acid in the metabolism of Schizosaccharomyces pombe: substrate consumption and cell growth

The effect of initial concentrations of malate varying from 0 to 28.6 g/l was studied. The acid was found to be inhibitory for growth of Schizosaccharomyces pombe but not for its deacidification activity. Malate was never integrated into biomass but partly transformed into ethanol if the aeration rate was weak (oxygen limitation). In the absence of glucose, resting cells of S. pombe were able to degrade malic acid if their concentration was sufficient, but their viability gradually decreased. However, for 0.15 g/l of growing cells (inoculum) 6 g/l of glucose was necessary to consume 8 g/l of malate. When the medium did not contain sugar no growth was observed despite the partial consumption of malate, showing that the acid was neither a carbon source nor an energy source

A model for pH determination during alcoholic fermentation of a grape must by Saccharomyces cerevisiae

A model to predict accurately pH evolution during alcoholic fermentation of must by Saccharomyces cerevisiae is proposed for the first time. The objective at least is to determine if the pH measurement could be used for predictive control. The inputs of the model are: the temperature, the concentrations in sugars, ethanol, nitrogen compounds, mineral elements (magnesium, calcium, potassium and sodium) and main organic acids (malic acid, citric acid, acetic acid, lactic acid, succinic acid). In order to avoid uncertainties coming from the possible precipitation, we studied this opportunity on a grape must without any tartaric acid, known as forming complexes with potassium and calcium during the fermentation. The model is based on thermodynamic equilibrium of electrolytic compounds in solution. The dissociation constants depend on the temperature and the alcoholic degree of the solution. The average activity coefficients are estimated by the Debbye–H¨uckel relation. A fictive diacid is introduced in the model to represent the unmeasured residual species. The molality of hydrogen ions and thus the pH are determined by solving a non-linear algebraic equations system consisted of mass balances, chemical equilibrium equations and electroneutrality principle. Simulation results showed a good capacity of the model to represent the pH evolution during fermentation

Malate utilization by Schizosaccharomyces pombe

Malic acid degradation bySchizosaccharomyces pombe was studied in synthetic and natural media. Using different concentrations from 1 to 29 g/l, malic acid was degraded. Initial deacidification rate increased with initial malate concentration. During the stationnary phase,S. pombe was able to decompose malate added to the medium before or after sugar exhaustion

Batch fermentation process: Modelling and direct sensitivity analysis

Based on a nonlinear model, this article realizes an investigation of dynamic behaviour of a batch fermentation process using direct sensitivity analysis (DSA). The used nonlinear mathematical model has a good qualitative and quantitative description of the alcoholic fermentation process. This model has been discussed and validated by authors in other studies. The DSA of dynamic model was used to calculate the matrix of the sensitivity functions in order to determine the influence of the small deviations of initial state, control inputs, and parameters from the ideal nominal values on the state trajectory and system output in time. Process optimization and advanced control strategies can be developed based on this work

Highlight on the problems generated by p-coumaric acid analysis in fermentations

p-Coumaric acid is a natural hydroxycinnamic acid existing in grapes and wine. It is the precursor of the 4-ethylphenol molecule through the bioconversion reaction by Brettanomyces yeast. Chromatographic methods are the most common techniques to detect p-coumaric acid. It is known that this acid is highly unstable in analysis and fermentation experiments. This paper highlights the problems occurring in p-coumaric acid analysis in wine fermentation conditions when studying its bioconversion. First, it was shown that p-coumaric acid was unstable at elevated temperature. On the other hand, it was found that in our experimental conditions p-coumaric acid reacted with ethanol. This work revealed also that the p-coumaric acid is partially adsorbed on Brettanomyces yeast, certainly on cell walls. Because of these phenomena the quantity of p-coumaric acid which can participate to the bioconversion into ethylphenol decreases

Malate degradation by Schizosaccharomyces yeasts included in alginate beads

Schizosaccharomyces yeasts can be used for deacidification of grape musts. To this aim, we studied malic acid degradation by yeasts included in double layer alginate beads in a bubble column reactor. Use of immobilized micro-organisms allowed a continuous process with high dilution rates giving a deacidification capacity of 0.032 g of malate/hour/dm3/g of beads. The pneumatic agitation was very convenient in this case

A direct and simple method for rapidly counting viable chains ofLeuconostoc in batch cultures

Plating determines the concentration in viable cells of bacteria but remains time-consuming and inaccurate. A direct and simple method combining two techniques, haemacytometry and epifluorescence, gives both the viability and the concentration in viable cells. The method was evaluated using Leuconostoc mesenteroides

Oxygen effect on batch cultures of Leuconostoc mesenteroides: relationship between oxygen uptake, growth and end-products

Growth and lactose metabolism of a Leuconostoc mesenteroides strain were studied in batch cultures at pH 6.5 and 30° C in 101 modified MRS medium sparged with different gases: nitrogen, air and pure oxygen. In all cases, growth occurred, but in aerobiosis there was oxygen consumption, leading to an improvement of growth yield Yx/s and specific growth rate compared to anaerobiosis. Whatever the extent of aerobic growth, oxygen uptake and biomass production increased with the oxygen transfer rate so that the oxygen growth yield, Yx/o2, remained at a constant value of 11 g dry weight of biomass/mol oxygen consumed. Pure oxygen had a positive effect on Leuconostoc growth. Oxygen transfer was limiting under air, but pure oxygen provided bacteria with sufficient dissolved oxygen and leuconostocs were able to consume large amounts of oxygen. Acetate production increased progressively with oxygen consumption so that the total molar concentration of acetate plus ethanol remained constant. Maximal Yx/s was obtained with a 120 l/h flow rate of pure oxygen: the switch from ethanol to acetate was almost complete. In this case, a 46.8 g/mol Yx/s and a 0.69 h−1 maximal growth rate could be reached