Effect of the addition of different quantities of amino acids to nitrogen-deficient must on the formation of esters, alcohols, and acids during wine alcoholic fermentation

In this work the effect of the addition of different quantities of amino acids to a nitrogen-deficient must on the formation of volatile compounds during the wine alcoholic fermentation was studied. To do so, fermentations of Mazuelo must were carried out, to which were added ammonium and 45, 120, and 250 mg/l of amino acids. The results showed that the formation of total esters, isoamyl acetate, and 2-phenylethyl acetate was directly proportional to the quantity of amino acids added to the must while the synthesis of diethyl succinate, and ethyl 3-hydroxybutyrate was inversely proportional to this addition. The alcohols, with the exception of tyrosol and 2-phenylethanol, did not show any direct correlation between their formation and the addition of amino acids. The quantity of amino acids added to the must favoured the formation of total acids but it had scant influence on the individual synthesis of the acids. So, it may be stated that, in general, the addition of amino acids in the concentrations under study to a nitrogen-deficient must, favoured the formation of volatile compounds in the wine.     

Addition of amino acids to grape juice of the Merlot variety: Effect on amino acid uptake and aroma generation during alcoholic fermentation

The effects of adding selected amino acids (phenylalanine, alanine, aspartic acid and threonine) to grape juice on the generation of aroma compounds and on amino acid uptake were studied. The fermentation kinetics varied according to the quantities of amino acids added. The fermentations finished more quickly in supplemented juices and their alcoholic content was significantly higher than in the control (p < 0.05). Amino acids were consumed mainly in the first quarter of fermentation. Higher alcohol formation took place at the same time as ethanol formation: with more amino acids present in the medium, more phenyl ethanol (p = 0.01) and benzyl alcohol were formed while isoamyl alcohol production decreased. The contents of isoamyl and phenylethyl acetates, ethyl hexanoate and ethyl octanoate, as well as most fatty acids increased during the fermentation, reaching a maximum for 10% of ethanol; with higher alcoholic contents, their concentrations decreased.     

Influence of SO2 on the consumption of nitrogen compounds through alcoholic fermentation of must sterilized by pulsed electric fields

The aim of this work was to study the influence of SO₂ on the use of nitrogenous compounds by yeast during wine alcoholic fermentation. Thus Parellada must was sterilized by a pulsed electric field treatment and inoculated with Saccharomyces cerevisiae Na33 strain. The fermentations were carried out with SO₂ (20 mg/l) and without SO₂. Results showed that yeast better consumed the amino acids in the first half of fermentation in the presence of SO₂. The final concentration of amino acids in the obtained wine was greater when the must was fermented without SO₂ than when the latter compound was present. Therefore, the presence of SO₂ facilitated the consumption of amino acids and, hence, such wine should have more complex flavour and better microbiological stability than that obtained from fermentation without SO₂.     

Optimization of honey-must preparation and alcoholic fermentation by Saccharomyces cerevisiae for mead production

Mead fermentation is a time-consuming process, often taking several months to complete. Despite of the use of starter cultures several problems still persist such as lack of uniformity of the final products, slow or premature fermentation arrest and the production of off-flavors by yeast. Thus the aim of this study was to optimize mead production through the use of an appropriate honey-must formulation to improve yeast performance alcoholic fermentation and thereby obtain a high quality product. Honey-must was centrifuged to reduce insoluble solids, pasteurized at 65 °C for 10 min, and then subjected to different conditions: nitrogen supplementation and addition of organic acids. Although the addition of diammonium phosphate (DAP) reduced fermentation length, it did not guarantee the completeness of the fermentation process, suggesting that other factors could account for the reduced yeast activity in honey-must fermentations. Sixteen yeast-derived aroma compounds which contribute to the sensorial quality of mead were identified and quantified. Global analysis of aromatic profiles revealed that the total concentration of aroma compounds in meads was higher in those fermentations where DAP was added. A positive correlation between nitrogen availability and the levels of ethyl and acetate esters, associated to the fruity character of fermented beverages, was observed whereas the presence of potassium tartrate and malic acid decreased, in general, their concentration.This study provides very useful information that can be used for improving mead quality.     

Sedimentation clarification of viura musts. Utilization of amino acids during fermentation

The influence of sedimentation on viura must's concentration of free amino acids and changes during fermentation were studied; the results were compared with an unclarified sample. The protein fraction was reduced by this treatment; the amino nitrogen increased slightly in the decanted must, and the concentration of ammonium nitrogen was similar in both samples. The decanted must had a greater concentration for most of the amino acid constituents of proteins, neutral as well as basic. This treatment did not alter the medium sufficiently to modify yeast utilization of these nutrients during fermentation; in fact, in both samples, basic, neutral, and acidic amino acids were consumed in a similar manner in the first half of fermentation (until 50% sugar consumption) and were excreted similarly in the freshly produced wine.     

Amino acid uptake by wild and commercial yeasts in single fermentations and co-fermentations

Musts require nitrogen-containing compounds in order to ensure yeast development. This study examined the nitrogen-nutrient requirements of two commercial yeasts and three wild strains isolated from inoculated fermentations. The results showed that wild strains generally consumed lower amounts of amino acids than commercial yeasts. Most amino acids were assimilated during the exponential growth phase; only a few – including asparagine and histidine – were metabolized until the end of fermentation. The study also sought to determine whether industrial drying affected yeast nitrogen requirements.     

Antioxidant phenolic compounds loss during the fermentation of Chétoui olives

Evolution of phenolic compounds was studied during spontaneous and controlled fermentations of “Chétoui” cultivar olives at three degree of ripeness. Both spontaneous and controlled fermentations led to an important loss of total phenolic compounds with a reduction rate of 32–58%. Consequently, the antioxidant activity decreased by 50–72%. After fermentations, hydroxytyrosol and caffeic acid concentrations increased, whilst protocatechuic acid, ferulic acid and oleuropein concentrations decreased. The hydroxytyrosol concentration in black olives increased from 165 mg/100 g dry weight to 312 and 380 mg/100 g dry weight, respectively, after spontaneous and controlled fermentation. The oleuropein concentration in green olives decreased from 266 mg/100 g dry weight to 30.7 and 16.1 mg/100 g dry weight, respectively, after spontaneous and controlled fermentation. During olive fermentation, phenolic loss is essentially due to the diffusion of these compounds into the brine; the main phenolic compound identified and quantified in the different brines was hydroxytyrosol. To preserve antioxidant quality of table olives it is necessary to use a controlled process to minimise phenolic compound loss.     

The effect of nitrogen source on yeast metabolism and H2S formation

The impacts of aspartic acid and glutamine used as nitrogen supplements for alcoholic fermentations conducted by Saccharomyces were studied. Synthetic grape juice media and commercially prepared grape juices were supplemented with diammonium phosphate, aspartic acid, or glutamine to increase yeast-assimilable nitrogen concentrations to 250 mg N/L prior to fermentation. Two yeast strains, UCD522 and EC1118, were inoculated at 10⁵ CFU/mL and fermentations were monitored for soluble solids, hydrogen sulfide (H₂S), and residual amino acids. In general, unsupplemented media/juices fermented slower than supplemented ones, produced more H₂S, and contained lower concentrations of amino acids after fermentation. Among the supplemented treatments, fermentation rates, H₂S production, and amino acid utilization varied depending on the nitrogen source and yeast strain. Those fermentations supplemented with aspartic acid were generally slower and sometimes did not achieve dryness. In contrast, glutamine additions yielded fermentation rates and H₂S production equivalent or better than other supplemented treatments. Based on these results, the use of glutamine appears promising as an alternative nitrogen supplement for wine production.     

Influence of addition of yeast autolysate on the formation of amines in wine

Amines, substances which can be toxic or have a negative effect on wine aroma, are formed during alcoholic and malolactic fermentations depending on various factors. In this work the influence of the addition of yeast autolysate to must on the concentration of amines in wine is studied. For this purpose Chardonnay must supplemented with the yeast autolysate was used; a second sample was also used without it (control sample). This autolysate provided amino acids and long‐chain fatty acids (C16 and C18). The enrichment of must by these nutrients did not produce an increase of amines during alcoholic fermentation despite the fact that consumption of amino nitrogen in general, and of some precursor amino acids in particular, was somewhat higher in the supplemented sample. After malolactic fermentation it was observed that the concentration of biogenic amines was higher in the wine from the supplemented must. Copyright © 2006 Society of Chemical Industry     

Effect of pure and mixed cultures of the main wine yeast species on grape must fermentations

Mixed inoculation of non-Saccharomyces yeasts and S. cerevisiae is of interest for the wine industry for technological and sensory reasons. We have analysed how mixed inocula of the main non-Saccharomyces yeasts and S. cerevisiae affect fermentation performance, nitrogen consumption and volatile compound production in a natural Macabeo grape must. Sterile must was fermented in triplicates and under the following six conditions: three pure cultures of S. cerevisiae, Hanseniaspora uvarum and Candida zemplinina and the mixtures of H. uvarum:S. cerevisiae (90:10), C. zemplinina:S. cerevisiae (90:10) and H. uvarum:C. zemplinina:S. cerevisiae (45:45:10). The presence of non-Saccharomyces yeasts slowed down the fermentations and produced higher levels of glycerol and acetic acid. Only the pure H. uvarum fermentations were unable to finish. Mixed fermentations consumed more of the available amino acids and were more complex and thus better able to synthesise volatile compounds. However, the amount of acetic acid was well above the admissible levels and compromises the immediate application of mixed cultures.     

Exploitation of Acha (Digitaria exiliis) and Iburu (Digitaria iburua) flours: Chemical characterization and their use for sourdough fermentation

This study aimed at characterizing the chemical and microbiological composition of Acha (Digitaria exiliis) and Iburu (Digitaria iburua) flours and at exploiting their potential for sourdough fermentation. Both the flours had a gross composition similar to other cereals. As shown by two-dimensional electrophoresis analysis, Acha flour had a higher number of prolamins with respect to Iburu flour. The opposite was found for the number of glutelin spots. The concentration of total free amino acids of Iburu flour was higher than that of Acha flour (1348 ± 34 vs. 100 ± 20 mg/kg). Pediococcus pentosaceus was the dominant species in both the flours. Several isolates were used to ferment Acha or Iburu flours. After 8 h fermentation at 30 °C, pH ranged from 3.41 to 3.83 and from 4.20 to 4.66 for Acha and Iburu sourdoughs, respectively. The highest values of TTA and concentration of lactic acid were found in almost all Iburu sourdoughs. The synthesis of acetoin and γ-amino butyric acid (GABA) was only found in Iburu sourdoughs. Data from the kinetics of acidification, synthesis of lactic and acetic acids, acetoin, and liberation of total free amino acids were elaborated by Principal Component Analysis. Sourdoughs from Acha and Iburu flour were clearly differentiated.     

Fermentation of grape must by sequential association of yeasts: Accumulation of pyruvic and α-ketoglutaric acid

Fermentations of grape (cv. Malvar) musts from two consecutive vintages were carried out using the autochthonous microflora, a sequential association of yeasts and conventional fermentations with addition of sulfur dioxide to the must. The pyruvic and α-ketoglutaric acid content over the course of fermentation was measured and showed that for both vintages tested the maximum accumulation of the ketoacid pyruvic acid took place several days earlier in fermentations using a sequential association of yeasts than in conventional fermentations. The accumulation of pyruvic acid was higher in the must made from grapes with a higher degree of ripening and the lowest level of added SO₂. In the fermentations using either a sequential association of yeasts or the autochthonous microflora with no added SO₂, accumulation of α-ketoglutaric acid was higher in the must with the higher nitrogen content when the species making the greatest percentage contributions at the start of fermentation presented high levels of proteolytic activity.     

Multifactorial analysis of acetaldehyde kinetics during alcoholic fermentation by Saccharomyces cerevisiae

Acetaldehyde is the terminal electron acceptor in the alcoholic fermentation by Saccharomyces cerevisiae. Quantitatively the most important carbonyl by-product, it has relevance for ethanol production yields as well as product stabilization and toxicology. The aim of this study was to investigate the effect of various enological parameters on acetaldehyde kinetics during alcoholic fermentations. Two commercial yeast strains were tested in two grape musts and the pH, temperature, SO₂ and nutrient addition were varied. All incubations had uniform kinetics where acetaldehyde reached an initial peak value followed by partial reutilization. Peak acetaldehyde concentrations and residual concentrations after 15 days of fermentations ranged from 62 to 119 mg l^− 1 and 22 to 49 mg l^− 1, respectively. A positive linear relationship was found between peak and final acetaldehyde levels in Gewürztraminer, but not Sauvignon Blanc fermentations, where sluggish fermentations were observed. Several factors had a significant effect on peak and/or final acetaldehyde levels. SO₂ addition, grape cultivar and fermentation nutrition were important regulators of peak acetaldehyde production, while final acetaldehyde concentrations were correlated with SO₂ addition, grape cultivar and temperature. The results allowed to estimate the acetaldehyde increase caused by SO₂ addition to 366 ??g of acetaldehyde per mg of SO₂ added to the must. The course of the final fermentation phase was shown to determine acetaldehyde residues. Comparison of acetaldehyde and hexose kinetics revealed a possible relationship between the time of occurrence of peak acetaldehyde concentrations and the divergence of glucose and fructose degradation rates.     

Formation of vinylphenolic pyranoanthocyanins by Saccharomyces cerevisiae and Pichia guillermondii in red wines produced following different fermentation strategies

The strains of two species of yeast, Saccharomyces cerevisiae and Pichia guillermondii, both with high hydroxycinnamate decarboxylase (HCDC) activity (56% and 90% respectively), were used in the fermentation of musts enriched with grape anthocyanins, to favour the formation of highly stable vinylphenolic pyranoanthocyanin pigments. The different strains were used to ferment the must separately, simultaneously, or sequentially, the latter involving an initial period using the yeast with the greatest HCDC activity (P. guillermondii). The must was made from concentrated grape juice diluted to 220 g/l of sugar, and enriched with grape anthocyanins to 100 mg/l and with p-coumaric acid to 120 mg/l. The pH was fixed to 3.5. All 50 ml micro-fermentations were done in triplicate. The development of anthocyanin-3-O-glucoside precursors, the decarboxylation of p-coumaric acid, and the formation of 4-vinylphenol and vinylphenolic pyranoanthocyanin derivatives were studied during the fermentation. The fermentation strategy used and the yeast HCDC activity significantly influenced the formation of vinylphenolic pyranoanthocyanins. The latter molecules were separated, identified, and quantified using high performance liquid chromatograph with diode array detection and electrospray-mass spectrometry (HPLC-DAD-ESI/MS). The volatile compounds profile was screened during fermentation using gas cromatogrphy-flame ionisation detection (GC-FID), in order to detect and quantify the main molecules. The best results were reached with the sequential fermentation (3.74 mg/l of malvidin-3-O-glucoside-4-vinylphenol). This work shows that during mixed or sequential fermentations carried out with non-Saccharomyces or highly fermentative Saccharomyces strains, with high HCDC activity, the content of stable pigments can be increased without sensorial modifications.     

Effect of ammonium nitrogen supplementation of grape juice on wine volatiles and non-volatiles composition of the aromatic grape variety Albariño

Vitis vinifera cv. Albariño is an aromatic variety characterised by terpenes present largely as glycosidic flavour precursor compounds, which depends on fermentation to reveal its aromatic varietal potential. Clarified Albariño must containing 250 mg N/L (control) was supplemented with diammonium phosphate to 350 and 450 mg N/L before fermentation with a high-nitrogen-demand wine strain of Saccharomyces cerevisiae (M05). Ammonium supplementation had a significant effect on the chemical composition (titratable acidity and ethanol) and volatile profile of Albariño wines. Varietal compounds in the free fraction (limonene, linalool, α-terpineol, α-ionone and β-damascenone) and bound (limonene, linalool and α-terpineol), as well as most yeast-derived fermentation products (including esters, higher alcohols and volatile acids) were associated with moderate nitrogen concentrations. Free β-ionone, bound geraniol, bound α-ionone, bound β-damascenone and 1-hexanol were associated with high nitrogen concentrations. OAV wines made with moderate nitrogen exhibited a higher total odorant activity, driven by ethyl esters, terpenes and C₁₃-norisoprenoids, resulting in fruity and floral aroma attributes.     

Novel starter cultures to inhibit biogenic amines accumulation during fish sauce fermentation

Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in food products such as meat and fish sausages. However, little information is available regarding the application of these bacteria in fish sauce. Hence, our study was aimed to investigate the effect of such starter cultures in reducing biogenic amines accumulation during fish sauce fermentation. Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05 isolated from fish sauce which possess amine oxidase activity were used as starter cultures in this study. Fermentation was held for 120 days at 35 °C. The pH value increased in all samples, while salt concentration remained constant throughout fermentation. Aerobic bacteria count was significantly lower (p < 0.05) in the control than in inoculated samples as a result of starter cultures addition. However, it decreased during fermentation due to the growth inhibition by high salt concentration. Proteolytic bacterial count decreased during fermentation with no significant difference (p > 0.05) among samples. These bacteria hydrolyzed protein in anchovy to produce free amino acid precursors for amines formation by decarboxylase bacteria. The presence of biogenic amines producing bacteria in this study was considered to be indigenous from raw material or contamination during fermentation, since our cultures were negative histamine producers. Amino acid histidine, arginine, lysine and tyrosine concentration decreased at different rates during fermentation as they were converted into their respective amines. In general, biogenic amines concentration namely histamine, putrescine, cadaverine and tyramine increased throughout fermentation. However, their concentrations were markedly higher (p < 0.05) in the control (without starter cultures) as compared to the samples treated with starter cultures. Histamine concentration was reduced by 27.7% and 15.4% by Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively. Both cultures could also reduce other amines during fermentation. After 120 days of fermentation, the overall biogenic amines concentration was 15.9% and 12.5% less in samples inoculated with Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively, as compared to control samples. These findings emphasized that application of starter cultures with amines oxidase activity in fish sauce fermentation was found to be effective in reducing biogenic amines accumulation.     

Nitrogen metabolism in chardonnay musts inoculated with killer strains of Saccharomyces cerevisiae

This paper examines the evolution of nitrogen compounds during alcoholic fermentation in chardonnay musts inoculated with killer strains of Saccharomyces cerevisiae (ICV D47 and ICV K1M). The results are compared with those obtained from chardonnay must fermented with indigenous yeasts (control sample). The total quantity of nitrogen assimilated, as well as the kinetics of utilization of ammonium ion and amino acids, was different depending on the yeast responsible for the fermentation. In the must inoculated with strain K1M, the nitrogen compounds were consumed more quickly and in greater quantity than in the must inoculated with strain D47. In the uninoculated must, the ammonium ion was not completely consumed, the consumption of amino acids was scant, the fermentation was slow, and the resulting wine had a high volatile acidity. Independently of the yeast used, in all the samples a preference was observed in the consumption of amino acids found in proteins over the amino acids not found in proteins and proline.     

Proximate composition and digestibility of fermented and extruded uji from maize-finger millet blend

The proximate composition, amino acid profile and in vitro starch and protein digestibilities of raw; fermented; fermented and cooked; unfermented and extruded; and fermented and extruded maize-finger millet blend was studied. Aspartic acid, glycine, cystine, methionine, tyrosine and lysine increased after fermentation, while contents of all other amino acids showed no significant changes. Greater losses of amino acids occurred when the fermented blend was extruded than when cooked. Fermentation improved protein and starch digestibilities, whereas cooking or extruding the fermented blend reduced the digestibilities. Extruding the unfermented blend increased protein and starch digestibilities and reduced nitrogen solubility index by 50%. Raw flour had 0.41 g/100 g water-soluble starch which declined to 0.05 g/100 g on fermentation but increased to 20-34 g/100 g after extrusion.     

A Blend of Essential Plant Oils Used as an Additive to Alter Silage Fermentation or Used as a Feed Additive for Lactating Dairy Cows

A blend of essential plant oils was evaluated for its effects on silage fermentation and animal performance. In the first experiment, the blend of essential oils was mixed with freshly chopped whole-plant corn to achieve a concentration of 0, 40, or 80 mg of active product per kilogram of fresh forage weight. Whole-plant corn was also mixed with a buffered propionic acid-based product at 0.2% of fresh forage weight. The blend of essential oils did not affect the populations of yeasts, molds, lactic acid bacteria, or enterobacteria; the fermentation end products; or the aerobic stability of the corn silage. Addition of the buffered propionic acid additive moderately reduced the production of acids during fermentation and resulted in a small reduction in the numbers of yeasts after ensiling, but did not affect aerobic stability. In a second experiment, 30 Holstein cows (4 primiparous and 26 multiparous) averaging 118 ± 70 d in milk and producing 38 ± 16 kg of milk/d were fed a total mixed ration, once daily, that consisted of (on a DM basis) 25% corn silage, 15% alfalfa silage, 10% alfalfa hay, and 50% concentrate. One-half of the cows were fed a blend of essential oils that was mixed directly into their total mixed ration to provide 1.2 g/cow per d for 9 wk. Cows fed the essential oils ate 1.9 kg more dry matter/d and produced 2.7 kg more 3.5% fat-corrected milk/d than did cows fed the control diet. The percentages of milk fat and protein, the somatic cell count numbers, and the concentrations of milk urea nitrogen were unaffected by treatment. Feed efficiency, change in body weight, and change in body condition scoring were also similar between treatments. After 12 h of incubation, the addition of a moderate dose and a high dose of essential oils to in vitro ruminal fermentations had no effect on the concentration of total VFA compared with the control treatment. However, they decreased the molar proportions of acetic, butyric, and valeric acids and increased the proportion of propionic acid. The blend of essential oils evaluated in this study altered in vitro ruminal fermentation and improved animal performance when fed directly to cows, but it did not affect the fermentation or aerobic stability of corn silage.     

以废弃毕赤酵母为高效氮源强化丁酸发酵生产

为有效利用固态废弃毕赤酵母,高效发酵生产丁酸,建立了以80 g/L玉米淀粉为基础培养基、以废弃毕赤酵母处理液为高效有机氮源,连续补加葡萄糖的丁酸发酵工艺。7 L厌氧发酵罐下,发酵20h,先以1∶4的比例添加250~400 g/L规格的废弃酵母处理液替代昂贵的发酵用复合培养基,再连续补加葡萄糖浓缩液。最终的丁酸质量浓度为45 g/L、得率为40%,丁酸占总酸的比率(butyric acid ratio over total organic acids,B/TA)≥0.90的较高水平。该工艺可以有效地利用来自于废弃酵母处理液中的氨基酸,提高细胞生长速度和浓度,增强丁酸合成所依赖的NADH再生速度,间接改善了丁酸发酵性能。与此同时,实现了废弃生物质的有效利用和资源化。