Free amino acids and volatile compounds in vinegars obtained from different types of substrate

The present work reports on the free amino acids and volatile compounds in vinegars obtained from different types of raw materials (cider, white wine and red wine). Based on the amino acid contents of three types of vinegar and of the substrates from which they were obtained, the profile of the nitrogen‐supplying compounds and their uptake for the acetic acid bacteria were very similar. The most important amino acid in terms of supply and uptake was found to be L ‐leucine. L ‐Proline proved also important in the wine vinegars, however it was not depleted as L ‐leucine. The different acetification reactions involved were found to yield acetoin, and plus 1,1‐diethoxyethane in the wine vinegars. Copyright © 2004 Society of Chemical Industry     

Effects of branched‐chain amino acid addition on chemical constituents in lychee wine fermented with Saccharomyces cerevisiae

This study investigated the impact of single branched‐chain amino acid addition (L‐valine, L‐leucine, L‐isoleucine) on nonvolatile and volatile constituents in lychee wine fermented with Saccharomyces cerevisiae MERIT.ferm. The individual addition of branched‐chain amino acids decreased the consumption of proline with significantly different rates. The lychee wine with added L‐leucine and L‐isoleucine had higher odour activity values (OAVs) of isoamyl alcohol, active amyl alcohol and isoamyl acetate. The lychee wine with added L‐valine had higher OAVs of isobutyric acid and isobutyl alcohol (5.55 and 2.98, respectively), which were lower in the wine with added L‐leucine (2.91 and 1.02, respectively) or L‐isoleucine (3.43 and 1.73, respectively) relative to the control wine (4.18 and 1.77, respectively), suggesting interactive effects among the amino acids. These findings demonstrated that lychee wine aroma can potentially be manipulated via the addition of selected branched‐chain amino acids.     

Free amino acid composition in primary and secondary inflorescences of 11 broccoli (Brassica oleracea var italica) cultivars and its variation between growing seasons

Eleven broccoli cultivars were grown in the field in spring/summer (April–July) and summer/winter (September–January). Free amino acid composition was determined by HPLC in primary and secondary inflorescences separately. A total of 17 amino acids were identified: L ‐alanine (Ala), L ‐arginine (Arg), L ‐asparagine (Asn), L ‐aspartic acid (Asp), glycine (Gly), L ‐glutamic acid (Glu), L ‐glutamine (Gln), L ‐histidine (His), L ‐isoleucine (Ile), L ‐leucine (Leu), L ‐methionine (Met), L ‐phenylalanine (Phe), L ‐serine (Ser), L ‐threonine (Thr), L ‐tryptophan (Trp), L ‐tyrosine (Tyr) and L ‐valine (Val). The major amino acid was L ‐glutamine, which represented on average between 39.5% (in cvs Durango and Green Valiant) and 55.5% (in cv Shogun) of the total amino acid content among cultivars, followed by L ‐glutamic acid with a variation between 12.1% (in cv Shogun) and 17.4% (in cv Marathon). A few amino acids represented less than 1% each (Gly, Leu, Met, Phen, Thr, Trp and Tyr). For most of the amino acids there were significant differences between cultivars, whilst only a few amino acids showed significant variations between inflorescences. Season also induced significant differences in the content of most of the identified amino acids. The cultivar with the highest total free amino acid content (323.9 mmol kg⁻¹ DW) on average of both seasons (391.3 in spring/summer and 256.4 in summer/winter) was Shogun, whilst the others were above the minimum of 177.3 mmol kg⁻¹ DW found in cv SK3. There was a general tendency for higher total amino acids levels in spring/summer than in summer/winter, but it was clear that this effect was dependent on the cultivar. © 2000 Society of Chemical Industry     

Biosynthesis of higher alcohol flavour compounds by the yeast Saccharomyces cerevisiae: impact of oxygen availability and responses to glucose pulse in minimal growth medium with leucine as sole nitrogen source

Higher alcohol formation by yeast is of great interest in the field of fermented beverages. Among them, medium‐chain alcohols impact greatly the final flavour profile of alcoholic beverages, even at low concentrations. It is widely accepted that amino acid metabolism in yeasts directly influences higher alcohol formation, especially the catabolism of aromatic and branched‐chain amino acids. However, it is not clear how the availability of oxygen and glucose metabolism influence the final higher alcohol levels in fermented beverages. Here, using an industrial Brazilian cachaça strain of Saccharomyces cerevisiae, we investigated the effect of oxygen limitation and glucose pulse on the accumulation of higher alcohol compounds in batch cultures, with glucose (20 g/l) and leucine (9.8 g/l) as the carbon and nitrogen sources, respectively. Fermentative metabolites and CO₂/O₂ balance were analysed in order to correlate the results with physiological data. Our results show that the accumulation of isoamyl alcohol by yeast is independent of oxygen availability in the medium, depending mainly on leucine, α‐keto‐acids and/or NADH pools. High‐availability leucine experiments showed a novel and unexpected accumulation of isobutanol, active amyl alcohol and 2‐phenylethanol, which could be attributed to de novo biosynthesis of valine, isoleucine and phenylalanine and subsequent outflow of these pathways. In carbon‐exhausted conditions, our results also describe, for the first time, the metabolization of isoamyl alcohol, isobutanol, active amyl alcohol but not of 2‐phenylethanol, by yeast strains in stationary phase, suggesting a role for these higher alcohols as carbon source for cell maintenance and/or redox homeostasis during this physiological phase. Copyright © 2014 John Wiley & Sons, Ltd.     

Concomitant extracellular accumulation of alpha-keto acids and higher alcohols by Zygosaccharomyces rouxii

Alpha-keto acids are key intermediates in the formation of higher alcohols, important flavor components in soy sauce, and produced by the salt-tolerant yeast Zygosaccharomyces rouxii. Unlike most of the higher alcohols, the alpha-keto acids are usually not extracellularly accumulated by Z. rouxii when it is cultivated with ammonium as the sole nitrogen source. To facilitate extracellular accumulation of the alpha-keto acids from aspartate-derived amino acid metabolism, the amino acids valine, leucine, threonine and methionine were exogenously supplied during batch and A-star cultivations of (routants of) Z. rouxii. It was shown that all alpha-keto acids from the aspartate-derived amino acid metabolism, except alpha-ketobutyrate, could be extracellularly accumulated. In addition, it appeared from the concomitant extracellular accumulation of alpha-keto acids and higher alcohols that in Z. rouxii, valine, leucine and methionine were converted via Ehrlich pathways similar to those in Saccharomyces cerevisiae. Unlike these amino acids, threonine was converted via both the Ehrlich and amino acid biosynthetic pathways in Z. rouxii.     

Release of macromolecules by Saccharomyces cerevisiae during ageing of French flor sherry wine "Vin jaune"

The French flor sherry wine "Vin jaune" spends 6 years and 3 months in the same barrel under a yeast velum. Because of temperature variations in the cellars, this velum sinks partially into the wine and a deposit of dead yeasts cells accumulates in the bottom of the barrels, favouring the formation of new velum. Growth and autolysis occur simultaneously. This study investigated the evolution of macromolecules released by yeasts during the ageing of "Vin jaune" in a model system closely simulating winemaking. It was observed that the release of macromolecules during the formation of the velums by living yeasts was low but greatly increased when the velums fell and yeast viability decreased. The release of macromolecules was then due to the autolysis of dead cells. Analysis of macromolecules during ageing revealed that they contained 73.3-78.5% neutral sugars and 6-7% proteins according to the ageing stage. Their amino acid composition did not change during ageing. A high content of serine and threonine commonly involved in O-glycosidic linkages present in yeast mannoproteins was observed. Throughout ageing, the mannose and glucose contents of macromolecules increased but the ratio of polymeric mannose to glucose decreased. Size exclusion chromatography showed that mannoproteins released in wine were partially hydrolysed by yeast beta-1,3-glucanases freed in wine.     

Production of Garcinia wine: changes in biochemical parameters,organic acids and free sugars during fermentation of Garcinia must

Garcinia wine was prepared by fermentation of ameliorated must of Garcinia xanthochymus using Saccharomyces cerevisiae. The present studies focused on changes in biochemical parameters (brix, pH, aldehydes, esters and alcohols), organic acids (reduction of oxalic acid), free sugars and antioxidant activities on fermentation of Garcinia must. The wine had higher amount of residual sugars contributing to the calorific value. The aldehydes and esters content in the final wine were 0.034% and 0.26%, respectively. There was reduction of citric acid and oxalic acid (antinutritional factor) and synthesis of aspartic acid and glutamic acid. Garcinia beverage was accepted on sensory analysis with high score for desirable attributes and overall quality with alcohol content of 6.1%. There was increase in total phenolics (0.039% gallic acid equivalent) and reducing power on fermentation but decrease in 2,2‐diphenyl‐1‐picrylhydrazyl radical scavenging activity.     

A COMBINATION OF GLUTAMINASE AND pH CONTROL PREVENTS THE NONENZYMATIC CONVERSION OF l-GLUTAMINE INTO l-2-PYRROLIDINE-5-CARBOXYLIC ACID IN FOOD PROCESSING

A strategy using Alicyclobacillus F‐62 glutaminase was established to minimize the nonenzymatic production of l ‐2‐pyrrolidine‐5‐carboxylic acid (l ‐pyroglutamic acid, savorless amino acid) and maximize the enzymatic production of l ‐glutamic acid (savorous [umami] amino acid) from l ‐glutamine in food‐protein processing. The nonenzymatic production rate of l ‐pyroglutamic acid was influenced by pH and temperature, and decreased over a pH range of 4–6 at 30C. Under optimum conditions (30C and pH 5.0), the l ‐glutamic and l ‐pyroglutamic acids obtained from 1 mM l ‐glutamine in the presence of glutaminase from Alicyclobacillus F‐62 were 0.96 and 0.03 mM, respectively. This novel method is useful for the efficient production of l ‐glutamic acid during food‐protein processing.     

陈酿期间无花果酒风味品质的变化

陈酿是提升果酒品质的一种重要手段。研究基于无花果酒的挥发性风味物质、游离氨基酸含量和感官特性,以0、2、4、6个月的陈酿无花果酒为研究对象,探究陈酿对无花果酒风味的影响。结果表明,随着陈酿时间的延长,酯类和醛类物质种类及相对含量均有所增加,然而醇类和酸类物质种类及相对含量均下降。电子鼻结果显示,陈酿0、2、4个月对果酒风味影响有明显区别,第4个月是风味变化的平衡点。陈酿过程中,无花果酒游离氨基酸总量和甜味氨基酸（苏氨酸、丝氨酸、脯氨酸、甘氨酸、丙氨酸、蛋氨酸、精氨酸）含量在第4个月达到最高值后呈现下降趋势,苦味氨基酸（缬氨酸、异亮氨酸、亮氨酸、酪氨酸、苯丙氨酸、赖氨酸、组氨酸）含量显著下降（P＜0.05）,鲜味氨基酸（天冬氨酸、谷氨酸）含量显著上升（P＜0.05）。皮尔森相关性分析方法进一步确证,陈酿通过提升澄清度、增加酯类和醛类化合物、释放鲜甜味氨基酸,以平衡果酒香气口感、提升风味品质。     

Application of Plackett–Burman experimental design for investigating the effect of wort amino acids on flavour‐active compounds production during lager yeast fermentation

Aroma‐active higher alcohols and esters are produced intracellularly in the cytosol by fermenting lager yeast cells, which are of major industrial interest because they determine aroma and taste characteristics of the fermented beer. Wort amino acid composition and their utilization by yeast during brewer's wort fermentation influence both the yeast fermentation performance and the flavour profile of the finished product. To better understand the relationship between the yeast cell and wort amino acid composition, Plackett–Burman screening design was applied to measure the changes in nitrogen composition associated with yeast amino acids uptake and flavour formation during fermentation. Here, using an industrial lager brewing strain of Saccharomyces pastorianus , we investigated the effect of amino acid composition on the accumulation of higher alcohols and volatile esters. The objective of this study was to identify the significant amino acids involved in the flavour production during beer fermentation. Our results showed that even though different flavour substances were produced with different amino acid composition in the fermentation experiments, the discrepancies were not related to the total amount of amino acids in the synthetic medium. The most significant effect on higher alcohol production was exercised by the content of glutamic acid, aromatic amino acids and branch chain amino acids. Leucine, valine, glutamic acid, phenylalanine, serine and lysine were identified as important determinants for the formation of esters. The future applications of this information could drastically improve the current regime of selecting malt and adjunct or their formula with desired amino acids in wort. Copyright © 2017 The Institute of Brewing & Distilling     

Apple Aminoacid Profile and Yeast Strains in the Formation of Fusel Alcohols and Esters in Cider Production

The amino acid profile in dessert apple must and its effect on the synthesis of fusel alcohols and esters in cider were established by instrumental analysis. The amino acid profile was performed in nine apple musts. Two apple musts with high (>150 mg/L) and low (<75 mg/L) nitrogen content, and four enological yeast strains, were used in cider fermentation. The aspartic acid, asparagine and glutamic acid amino acids were the majority in all the apple juices, representing 57.10% to 81.95%. These three amino acids provided a high consumption (>90%) during fermentation in all the ciders. Principal component analysis (PCA) explained 81.42% of data variability and the separation of three groups for the analyzed samples was verified. The ciders manufactured with low nitrogen content showed sluggish fermentation and around 50% less content of volatile compounds (independent of the yeast strain used), which were mainly 3‐methyl‐1‐butanol (isoamyl alcohol) and esters. However, in the presence of amino acids (asparagine, aspartic acid, glutamic acid and alanine) there was a greater differentiation between the yeasts in the production of fusel alcohols and ethyl esters. High contents of these aminoacids in dessert apple musts are essential for the production of fusel alcohols and most of esters by aromatic yeasts during cider fermentation.     

Free amino acid profile during ripening of ewe's milk cheese from the Croatian island Krk

The aim of this study was to determine the content of free amino acids (FAA) and their ratio in ewe's milk cheese from the island Krk during its ripening. FAA content was determined by reversed phase HPLC (RP‐HPLC) of cheese aqueous/ethanol extracts after FAA were transformed into their 6‐aminoquinolyl‐N‐hydroxysuccinimidyl carbamate derivatives. Their concentration increased during ripening, reaching the value of 5% in cheese dry matter. The dominant FAA were glutamic acid>leucine>valine>aspartic acid>phenylalanine>serine>proline, and higher content of nonessential vs essential FAA was revealed. Krk cheese has, in relation to other cheeses, higher values for glutamic acid/leucine, glutamic acid/phenylalanine, glutamic acid/proline and smaller values for leucine/aspartic acid, valine/aspartic acid, phenylalanine/aspartic acid ratios, while other ratios are comparable to those of other hard ovine cheeses.     

Nutritional studies on agadagidi

The analysis showed a gradual fall of the sugar content as fermentation period increased. Ascorbic acid content decreased while nitrogen and crude protein increased during fermentation. The predominant amino acids are leucine, lysine, valine, asparagine and glutamic acid, methionine, phenylalanine.     

荔枝酒酿造过程中氨基酸的变化

利用高效液相色谱法(HPLC)对不同发酵温度的荔枝酒酿造过程中氨基酸含量的动态变化进行了分析研究。结果表明,荔枝汁和荔枝酒中均含有丙氨酸、苏氨酸、缬氨酸、谷氨酸、丝氨酸等14种氨基酸,其中,丙氨酸是荔枝汁和荔枝酒中的主体氨基酸,含量分别达1 139.682 mg/L、341.049～412.161 mg/L。不同发酵温度的荔枝酒酿造动态过程中,其游离氨基酸的种类和含量变化较大,且变化趋势各异。不同贮藏方式对荔枝酒中氨基酸含量和种类影响也不同。通过比较,发酵温度12℃优于15℃,发酵温度为15℃的荔枝酒中氨基酸含量在贮藏过程中变化幅度较大,不稳定,且影响风味的氨基酸较多。     

Higher alcohols concentration and its relation with the biological aging evolution

The production of 1-propanol, isobutanol, 2-methyl-1-butanol, 3-methyl-1-butanol and 2-phenylethanol by two flor yeast strains during accelerated biological aging of sherry wines with different aging time has been evaluated. The S. cerevisiae G1 strain has shown a higher production of higher alcohols than the S. bayanus F12 strain highlighting the 1-propanol production in the youngest wine. Differences between the two studied yeasts decreased in relation with the aging time of the initial wine used. The multiple regression analysis carried out to relate the higher alcohols concentration and the values obtained from an aging equation showed R-squared values ranging from 81.1 to 96.5%. The higher alcohols that best described such relation were 1-propanol, isobutanol and 3-methyl-1-butanol.     

Manipulation of volatile compound transformation in durian wine by nitrogen supplementation

This study evaluated the impact of added diammonium phosphate (DAP) and a L‐leucine‐L‐phenylalanine mixture (Leu+Phe) on durian wine fermentation by Saccharomyces cerevisiae var. bayanus EC‐1118. Changes in yeast cell population, ^oBrix, sugars, organic acids and pH were similar in all fermentations, regardless of nitrogen supplementation at a concentration of 150 mg N L^?1. The supplementation of Leu+Phe accelerated the utilisation and reduced the formation of volatile sulphur compounds such as diethyl disulphide, ethyl thioacetate and 2‐(ethylthio)ethanol, which were 64.3%, 26.0% and 48.4% lower than the control, respectively. Nevertheless, the supplementation of Leu+Phe heightened the production of isoamyl alcohol, 2‐phenylethyl alcohol and their corresponding esters, especially isoamyl acetate was 1.6 times and 2‐phenylethyl acetate was 26.5 times higher than the control. The supplementation with DAP exerted negligible effects on the volatile composition. The results of this study suggest that the addition of specific amino acids may be a novel approach to manipulating durian wine flavour by suppressing or accentuating the formation of certain aroma compounds.     

Effects of a mutated yeast plus addition of sucrose and nitrogen on the total higher alcohol levels of a plum wine fermentation

The brewing process and strain employed can play key roles in controlling the level of total higher alcohols (THA) in plum wine. In this study, fermentations using Saccharomyces cerevisiae XN3 and plum must led to the production of 671 mg/L THA (including 1‐propanol, isobutanol and isopentanol). A mutant of XN3, exhibiting a higher activity of lactic acid dehydrogenase, named XN3‐3, produced lower levels of THA at 538 mg/L. Further optimization of the brewing process, including rapid nitrogen and sugar fed‐batch additions, were conducted with XN3‐3. Tests with the addition of 0.4 g/L (NH₄)₂HPO₄ produced a lower level of THA (385 mg/L) than sucrose additions (to a 20°Brix level) using the fed‐batch method (452 mg/L). These additions were effective in reducing the higher alcohols and could be combined in application. Copyright © 2014 The Institute of Brewing & Distilling     

Determination of endogenous amino acid flow at the terminal ileum of the rat

In a comparative study endogenous amino acid flow was determined in twenty-four 190-g male rats (protein-replete) given diets containing synthetic amino acids as the sole nitrogen source but devoid of specific amino acids, and six rats fed a protein-free diet. Endogenous flows were not significantly different (P > 0.05) for aspartic acid, glutamic acid, serine and lysine but were significantly higher (P < 0.05) under protein-free alimentation than amino acid alimentation for proline, glycine and alanine. This indicates that the protein-free method did not lead to lowered endogenous amino acid excretions owing to an altered amino acid metabolism in the protein-free state. The loss of some amino acids may be enhanced under protein-free alimentation. The high apparent digestibility of the synthetic amino acids not excluded from the diets (histidine, arginine, threonine, valine, isoleucine, leucine, tyrosine, phenylalanine) indicated that their ileal excretions were mainly of endogenous origin. There were no significant differences (P>0.05) between the latter excretions and those obtained by feeding the rats a protein-free diet.     

Fatty Acid Content,Flavor Compounds,and Sensory Quality of Pork Loin as Affected by Dietary Supplementation with l‐arginine and Glutamic Acid

To investigate the effect of functional amino acid on meat flavor and eating quality, 60 growing‐finishing pigs (Duroc × Large White × Landrace) were dietarily supplemented with or without 1.0% l ‐arginine, glutamic acid, or l ‐arginine plus glutamic acid for 2 months. After animals were slaughtered, the muscle fatty acid profile, flavor compounds, and meat sensory quality were comparatively investigated. The results showed that dietary supplementation with arginine, glutamic acid, or arginine plus glutamic acid had little effect on free amino acids, no effect on 5′‐nucleotides and meat sensory taste traits, but supplementation with arginine plus glutamic acid significantly increased (P < 0.05) fat accumulation and fatty acid content in muscle, increased (P < 0.05) the formation of multiple fatty acid oxidation‐derived volatile compounds, and improved the tenderness, juiciness, and overall eating quality of meat. This study revealed that dietary supplementation with 1.0% l ‐arginine and glutamic acid could be used to improve meat eating quality in pork production.     

Effects of nitrogen catabolite repression‐related amino acids on the flavour of rice wine

The quality of rice wine is highly dependent on the content of the flavour compounds produced by the budding yeast Saccharomyces cerevisiae. In this study, the effects of three amino acids (arginine, glutamate and glutamine) related to nitrogen catabolite repression on the formation of flavour compounds were investigated. Each of these amino acids could promote the growth of S. cerevisiae, and a total of 83 flavour compounds were found in a model system of rice wine production. The effects of arginine, glutamate and glutamine on the content of the higher alcohols, amino acids and esters were significant, whereas the effects on the aldehydes and organic acids were slight. The results of this study could facilitate the development of new strategies to control the flavour pattern and improve the quality of rice wine. Copyright © 2015 The Institute of Brewing & Distilling