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.     

Characterisation of volatile compounds in an alcoholic beverage produced by whey fermentation

An alcoholic beverage (35.4% v/v ethanol) was produced by distillation of the fermented broth obtained by continuous whey fermentation with a lactose-fermenting yeast Kluyveromyces marxianus. Forty volatile compounds were identified in this drink by gas chromatography. Higher alcohols were the most abundant group of volatile compounds present, with isoamyl, isobutyl, 1-propanol, and isopentyl alcohols being found in highest quantities (887, 542, 266, and 176 mg/l, respectively). Ethyl acetate had the highest concentration (138 mg/l) among the esters. Besides higher alcohols and esters, other components, including aldehydes, acids and terpenes were also identified in the whey spirit. Considering that the quality of an alcoholic beverage can be evaluated by the relation between isoamyl alcohol/2-methyl-1-propanol and 2-methyl-1-propanol/1-propanol, which have to be higher than unity, it was concluded that a novel spirit of acceptable organoleptic characteristics can be produced by cheese whey continuous fermentation with K. marxianus.     

有机酸含量对葡萄酒发酵的影响

为了考察有机酸含量对葡萄酒发酵的影响,在初始有机酸含量分别为4 g/L、7 g/L和10 g/L的模拟葡萄汁中接种酵母,于25 ℃发酵,比较不同初始有机酸含量对葡萄酒发酵过程中酵母生长、降糖、有机酸及挥发性化合物含量的影响。结果表明,初始有机酸含量7 g/L时酵母生长和耗糖最慢,乙醇和乙醛产生量最低;随初始有机酸含量升高,pH值、酒石酸、苹果酸、乳酸、乙酸、总酸、乙酸乙酯和总酯含量呈下降趋势;琥珀酸、正丙醇、异丁醇、异戊醇及总高级醇含量呈上升趋势;但对β-苯乙醇、正丁醇及丁酸乙酯含量影响较小。初始有机酸含量10 g/L的葡萄酒发酵过程酵母生长快、降糖最快,产酸含量适中,同时香气较好。因此,葡萄酒发酵的适宜初始有机酸含量为10 g/L。     

绿原酸对酿酒酵母发酵特性的影响

以模拟苹果汁为发酵体系,添加质量浓度为0（对照组）、0.01、0.1、1.0 g/L的绿原酸,通过考察发酵过程中酿酒酵母CICC31084的生物量、葡萄糖消耗、CO2释放、发酵液乙醇生成以及发酵液关键香气物质变化,研究发酵过程中绿原酸对酿酒酵母发酵特性的影响。结果表明：高质量浓度绿原酸能够促进酵母细胞生长,加快CO2释放和糖代谢速率,延缓发酵前期乙醇生成速率,高质量浓度（1.0 g/L）绿原酸对酿酒酵母发酵特性的影响更加显著,能够缩短整个发酵周期2～3 d。绿原酸作用下,发酵末期香气物质苯乙醇、乙酸乙酯、乙酸异戊酯含量增加,正己酸乙酯、癸酸乙酯含量降低,而辛酸乙酯含量差异不大。     

Sake: Production and flavor

Abstract

Sake, its history in Japan, the unique brewing process, and microbes concerned with the characteristics of flavor are described. The main flavor components derived mainly from the fermentation process are higher alcohols, isoamyl acetate, ethyl caproate, and phenethyl acetate. These are the compounds which give an accent to the flavor of sake. The esters are formed mainly by yeast during mash fermentation. Isoamyl acetate is produced by the reaction of acetyl CoA with isoamyl alcohol catalyzed by alcohol acetyl transferase. The enzyme, bound to the yeast cell membrane, is unstable to heat and unsaturated fatty acids. The ester formation is regulated by the amount of isoamyl alcohol produced. Acyl CoA alcohol acyl transferease catalyzes the formation of ethyl caproate from caproyl CoA and ethanol. In this reaction, the amount of caproyl CoA supplied is important. The mechanism of higher alcohol formation, including the biosynthetic pathway of amino acids and its feedback regulation, is discussed. Finally, breeding and the practical use of sake yeast with high productivity of higher concentrations of higher alcohols and esters are described.     

静态顶空气质联用法测定霞多丽在酒精发酵过程中香气的变化

运用静态顶空-气相色谱质谱联用法,从霞多丽葡萄汁到酒精发酵结束,每隔12h对其香气成分进行检测。检测得到的香气成分通过质谱谱库进行定性,色谱峰面积归一化法进行定量。测定结果显示:霞多丽在酒精发酵过程中共检测出31中香气物质,其中醇类物质有11种,酯类物质有13种,其他物质(酸、醛、酮)有7种。发酵前后香气成分的种类和相对含量变化显著。其中酯类和醇类物质的种类和相对含量随着发酵的进行有所增加,而其他类(酸、醛、酮)物质的种类和相对含量随着发酵的进行急剧下降。发酵结束后,发酵液的主要香气成分为:乙醇、2-甲基丙醇、3-甲基丁醇、2,3-丁二醇、苯乙醇、正丙醇、己醇、乙酸己酯、辛酸乙酯、壬酸乙酯、2-苯基乙酸乙酯、十二酸乙酯、乙酸乙酯、2-甲基乙酸丙酯、3-甲基乙酸丁酯、己酸乙酯、乙酸。     

Influence of addition of ammonium and different amino acid concentrations on nitrogen metabolism in spontaneous must fermentation

The effect of the addition of different amino acid concentrations in must on yeast nitrogen metabolism during alcoholic fermentation was studied. To do this, fermentations of Mazuelo must, poor in nitrogen compounds, were carried out. Ammonium and different concentrations of amino acids (0, 45, 120, 250 and 450 mg/l) were added to the must. Addition of 45, 120 and 250 mg/l of proteic amino acids to the must increased the rate of fermentation. Proline was mainly consumed in fermentations with smaller amounts of amino nitrogen and, at the same time, this amino acid showed the highest residual concentration in the final wines. The consumption of other proteic amino acids was directly proportional to their concentration in the musts, with the exception of leucine and isoleucine that were synthesized. However, a difference in the percentages of the amino acids consumed by the yeasts was observed. The percentages of aspartic acid, alanine and arginine consumed were higher in the fermentations supplemented with amino acids than in the fermentation where only ammonium was added. The percentages of tyrosine and phenylalanine consumed gradually increased with increase of their initial concentration.     

低氮对酿酒酵母发酵过程中酯类物质合成的影响

采用顶空固相微萃取-气相色谱-质谱法（HS-SPME-GC-MS）测定了不同程度低氮胁迫下的模拟葡萄汁酒精发酵过程中酯类物质的含量。结果表明,低氮并没有改变各种酯类物质的生成规律,但影响了其发酵结束时酯类物质的含量。在发酵过程中,乙酸乙酯、己酸乙酯、辛酸乙酯和癸酸乙酯的含量逐渐增加;丁酸乙酯的含量在发酵第8天达到最大值后逐渐降低;乙酸异戊酯的含量先增加后在发酵第12天时达到最低值后又迅速增加;乙酸苯乙酯的含量在发酵前期（发酵4～6 d）迅速达到一个峰值,随后其含量缓慢降低;低氮胁迫下酯类物质的总量降低,其中乙酸苯乙酯和癸酸乙酯的含量增加,乙酸乙酯、乙酸异戊酯、丁酸乙酯和己酸乙酯的含量降低。因此,在葡萄酒的生产过程中可适当提高氮源浓度来提高酯类物质含量。     

Effects of Fusarium infection on the amino acid composition of winter wheat grain

Winter wheat (susceptible cultivar Ritmo) was grown in 2006 near Kiel and in 2007 near Heide in northern Germany. Plants were inoculated at anthesis using a Fusarium graminearum macroconidial suspension. The percentage of Fusarium-damaged kernels (FDK) ranged from 0 ± 2% to 28 ± 2%. The contents of the Fusarium mycotoxin deoxynivalenol (DON) and wheat amino acids were determined in the grain. Levels of the amino acids alanine, lysine, and tyrosine increased with the percentage of FDK or DON contents whereas glutamic acid contents decreased. Aspartic acid and threonine were not related to the percentage of FDK or DON contents. Effects of Fusarium infection on other amino acids were significant only at the sampling site with the higher degree of Fusarium-damage. Interestingly, those amino acids that increased consistently and significantly with the degree of Fusarium-damage are derived from phosphoenolpyruvate or pyruvate, suggesting that pathogen-induced changes in the glycolytic input for amino acid biosynthesis play a significant role for the amino acid composition of Fusarium-damaged winter wheat grain. On average, amino acid contents decreased by 0.13% compared to the amino acid content of sound kernels upon an increase of 1% of FDK.     

Seasonal amino acid profiles and mineral contents of green tiger shrimp (Penaeus semisulcatus De Haan, 1844) and speckled shrimp (Metapenaeus monoceros Fabricus, 1789) from the Eastern Mediterranean

The amino acid and mineral contents of green tiger shrimp (Penaeus semisulcatus) and speckled shrimp (Metapenaeus monoceros) from the North Eastern Mediterranean were determined for each season of the year. For both species, the highest protein values were recorded during the summer (p < 0.05). The most abundant amino acids were, in decreasing order, glutamic acid, aspartic acid, arginine, lysine, leucine, glycine and alanine. The ratio of essential to nonessential amino acids (mean value) for green tiger shrimp and speckled shrimp were 0.60 and 0.59, respectively. Mineral contents of the two species varied with season (p < 0.05), except for the Ca content in green tiger shrimp.     

Changes of phenolic acids and antioxidant activities during potherb mustard (Brassica juncea, Coss.) pickling

Phenolic acids in potherb mustard (Brassica juncea, Coss.) were determined and the effects of pickling methods on the contents of total free phenolic acids, total phenolic acids, total phenolics, and antioxidant activities were investigated. Gallic acid, protocatechuic acid, p-hydroxybenzoic acid, vanillic acid, caffeic acid, p-coumaric acid, ferulic acid, and sinapic acid were identified in the present study. The contents of total free phenolic acids, total phenolic acids and total phenolics in fresh potherb mustard were 84.8 ± 0.58 μg/g dry weight (DW), 539 ± 1.36 μg/g DW, and 7.95 ± 0.28 mg/g DW, respectively. The total free phenolic acids increased during the pickling processes, but the total phenolic acids, total phenolics, and antioxidant activities decreased. However, after 5 weeks of fermentation, all the pickling methods retained over 70% of total phenolic contents and above 65% of antioxidant capacities. The results indicated that pickling processes were relatively good methods for the preservation of phenolic acids and antioxidants for potherb mustard.     

Nitrogen addition influences formation of aroma compounds,volatile acidity and ethanol in nitrogen deficient media fermented by Saccharomyces cerevisiae wine strains

The effects of nitrogen addition into nitrogen deficient/depleted media on the release of aroma compounds post-fermentation were investigated in three commercial yeast strains of Saccharomyces cerevisiae which highlight the yeast strain effect as well as nitrogen effects. By comparing the two timings of nitrogen addition, prior to fermentation or later at stationary phase (72 h), it was shown that nitrogen addition at stationary phase significantly decreases ethanol and acetic acid formation and significantly increases the following compounds: 2-phenylethanol, ethyl isobutyrate, 2-phenylethyl acetate, ethyl 2-methylbutyrate and ethyl propionate in the three strains, and also isovaleric acid, isoamyl alcohol and ethyl isovalerate in both PYCC4072 and UCD522. The strain EC1118 produced significantly less medium chain fatty acids, hexanoic, octanoic and decanoic acids and their respective esters after nitrogen addition. Therefore, timing of nitrogen addition to a ferment media can vary the concentration of certain aroma compound and might provide a means for varying wine composition.     

Comparison of free amino acids and 5′-nucleotides between Tuber fermentation mycelia and natural fruiting bodies

The profiles of free amino acids and 5′-nucleotides were first compared between Tuber fermentation mycelia and natural fruiting bodies. A total of 20 free amino acids and five 5′-nucleotides were identified in the Tuber fermentation mycelia and natural fruiting bodies. Not only the total contents of the free amino acids and 5′-nucleotides, but also the contents of umami amino acids and flavour 5′-nucleotides in the fermentation mycelia were higher than those in the fruiting bodies. By the addition of soybean flour in the fermentation media, the flavour 5′-nucleotides content in the fermentation mycelia was significantly increased, and the equivalent umami concentration of the fermentation mycelia (i.e., 608.07 g/100 g) was approximately 38.1-93.4 times higher than those of the fruiting bodies. From the viewpoint of umami taste, this work confirms the potentiality of Tuber fermentation mycelia as the alternative resource for its fruiting bodies.     

李子果酒主发酵过程中理化指标及挥发性成分变化分析

该研究对李子果酒主发酵过程中的理化指标进行测定,并采用顶空固相萃取-气相色谱-质谱（HS-SPME-GC-MS）联用技术对其挥发性成分进行分析。结果表明,在李子果酒主发酵过程中,随着发酵时间的延长,pH值、酒精度呈先升高后趋于稳定的趋势;总酸、总糖含量呈先下降后趋于稳定的趋势;挥发酸含量呈上升趋势。主发酵结束后,李子果酒的酒精度为11.10%vol、总酸含量为9.13 g/L、pH值为3.76、挥发酸含量为0.25 g/L、总糖含量为22.90 g/L。主要变化的挥发性物质为6种醇类（乙醇、1-己醇、异戊醇、异丁醇、顺式-3-己烯-1醇、苯基乙醇）和6种酯类（乙酸乙酯、乙酸异戊酯、辛酸乙酯、乙酸己酯、己酸乙酯、癸酸乙酯）。通过对李子果酒主发酵过程动态变化分析,为提高李子果酒品质提供理论依据。     

菠萝皮渣糯米果酒发酵过程中主要成分变化研究

以菠萝皮渣和糯米为原料,进行糯米果酒发酵,并采用相关国标检测方法、液相色谱（LC）及气相色谱质谱联用（GC-MS）法,分析菠萝皮渣糯米果酒在发酵过程中可溶性固形物、酒精度、有机酸、挥发酸和香气成分等成分变化。结果表明：发酵过程中糖度迅速降低、酒精度升高后维持稳定,pH值在3.50～3.80之间,呈先降后升趋势,酸度先降低直至陈酿后期轻微升高。有机酸含量在发酵过程中总体呈下降趋势,草酸、酒石酸含量逐渐降低,柠檬酸、苹果酸、丙酮酸、乳酸含量先降后升,琥珀酸和乙酸呈上升趋势。果酒中挥发酸含量不断下降,第60天甲酸、乙酸含量分别为0.30 mg/L和0.26 mg/L。菠萝皮渣糯米果酒中共检测出88种香气成分,其中41种酯类、22种醇类、10种酸类和15种其他类,发酵过程中乳酸乙酯、乙酸异戊酯、2-甲基丁酸乙酯、正己酸乙酯、辛酸乙酯、乙酸苯乙酯、癸酸乙酯、月桂酸乙酯和十六酸乙酯含量较高,相对含量在0.27%～20.57%。     

Effect of local processing methods (boiling,frying and roasting) on the amino acid composition of four marine fishes commonly consumed in Nigeria

The effects of different processing methods on the amino acid contents of four marine fishes commonly consumed in Nigeria: Clupea harengus, Scomber scombrus, Trachurus trachurus and Urophycis tenuis were investigated. The samples were boiled, roasted over hot charcoal, or fried in any of four different oil types (palm oil, soybean oil, vegetable oil and groundnut oil). The total amino acids for all the fish samples ranged from 76.72 to 98.01 g/100 g protein and the total essential amino acids from 28.95 to 44.73 g/100 g protein. Frying reduced the total amino acid contents, with the lowest values obtained in samples fried in palm oil. Boiling and roasting increased the total amino acid contents, with the values higher in roasted than boiled samples. The percentage of essential amino acids reduced with processing in almost all samples, but the reduction was more pronounced in the fried samples. The raw, boiled and roasted samples had amino acid scores greater than 100%, while for the fried samples, the limiting amino acids were either leucine or threonine. The results showed that roasting and boiling had more desirable effects on the amino acid content of the fish samples.     

传统工艺山西老陈醋发酵及熏蒸过程中常规成分的变化分析

该研究分析了山西老陈醋在传统工艺发酵过程中淀粉酶、糖化酶、酸性和中性蛋白酶酶活变化,并对发酵及熏蒸过程中淀粉、总糖、酒精度、总酸、有机酸和氨基酸等常规成分的变化规律与形成机理进行了探讨。结果表明,酒精发酵阶段,4种酶活力变化不大,醋酸发酵阶段,4种酶活力均有所降低;发酵及熏蒸过程中淀粉含量由64.05%降至18.88%;酒精发酵阶段,总糖含量从11.88%降至1.12%,醋酸发酵阶段,总糖含量从2.28%增至7.35%,熏蒸阶段,总糖含量从7.13%降至5.89%;酒精含量在酒精发酵过程中从1.3%vol增至5.7%vol,到发酵22 d时几乎降为0;总酸含量在1～22 d内从1.17%增至10.23%;在发酵及熏蒸过程中醋酸含量从1.03 mg/g增至76.25 mg/g;酒精发酵阶段,乳酸含量从1.94 mg/g增至60.60 mg/g,随后含量变化较小。     

初始酸度对酿酒酵母发酵的影响

以高粱糖化液模拟发酵体系,添加0～30 mmol/100 g的乳酸以设置不同的初始发酵酸度,通过考察酿酒酵母（Saccharomyces cerevisiae）的生物量、发酵累计质量损失、发酵液乙醇生成、发酵液关键香气物质,探究不同初始酸度对酿酒酵母发酵状况的影响,以期解析初始酸度对白酒发酵产酒的影响。结果表明,随初始发酵酸度增加,酿酒酵母生长受到抑制,低初始酸度条件下（0～15 mmol/100 g）有利于酿酒酵母发酵,加快CO2释放和糖代谢速率及更多种类挥发性风味物质生成,当初始酸度为15 mmol/100 g时,醇类、酸类及总挥发性物质种类数最高;较高的初始酸度（15～30 mmol/100 g）会抑制酿酒酵母乙醇的生成。6种关键性挥发性风味物质检测结果表明,初始酸度为5 mmol/100 g时,酿酒酵母发酵末期的关键风味物质乳酸乙酯、苯乙醇、异丁醇和异戊醇含量达到最高,20 mmol/100 g的初始酸度更有利于乙酸乙酯生成,乙酸苯乙酯的生成随初始酸度增加而被抑制。综上,控制初始酸度为15 mmol/100 g左右发酵效果较好。     

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.     

外源添加单一氨基酸对桑葚酒风味品质的影响

为探究外源添加氨基酸在改善桑葚酒风味品质中的影响,在桑葚果汁中分别添加400 mg/L的异亮氨酸（Ile）、亮氨酸（Leu）、苯丙氨酸（Phe）和缬氨酸（Val）制备桑葚酒,并分析桑葚酒理化指标、挥发性风味物质和感官品质。结果表明,与对照相比,外源添加单一氨基酸可显著增加桑葚酒的挥发性风味物质含量（42.38～84.58 mg/L）（P＜0.05）,添加Ile显著增加活性戊醇、乙酸活性戊酯含量,添加Leu显著增加异戊醇、乙酸异戊酯含量,添加Phe显著增加苯乙醇和乙酸苯乙酯的含量,添加Val显著增加异丁醇和乙酸异丁酯含量,添加四种氨基酸都能增加辛酸乙酯的含量（P＞0.05）;添加Phe可提高桑葚酒花香,添加Ile、Leu或Val提高果香,添加四种氨基酸的桑葚酒的口感评分均显著提高（P＜0.05）。