啤酒酵母发酵产有机酸的生理代谢机制

以微波破壁和高效液相色谱法,对通风发酵过程中的啤酒酵母细胞胞外、胞内六种有机酸含量的动态变化进行了跟踪检测。研究结果表明,酵母细胞对一部分有机酸有着非常亲缘性的代谢途径和生理机制;柠檬酸等某些有机酸在酵母细胞衰老凋亡时作为碳底物代谢,存在着非常严格的精确保守性、经济效能性调控机制。     

MS-HPLC法检测啤酒酵母胞内代谢有机酸的变化

通过啤酒酵母细胞的几种破壁方法比较 ,确定了微波破壁 (MS ,microwave -split)和高效液相色谱(HPLC)联用的方法 ,对通风发酵过程中啤酒酵母细胞胞内六种有机酸含量的动态变化 ,进行了定性定量跟踪检测。本方法具有前处理简单、干扰小、分析速度快等优点 ,便于及时跟踪测定发酵过程中有机酸的动态变化。研究结果表明通风发酵过程中 ,胞内作为经济性碳底物的柠檬酸和琥珀酸含量很低 ,代谢变化调控严格 ;其余四种有机酸在发酵后期阶段有机酸含量变化对K 、Ca2 两种离子响应不同     

离子对啤酒酵母代谢产酸的影响

在离子型培养基中分别添加6.804,13.610,27.216 g/L 的KH2PO4以及66.39,138.75, 277.4 mg /L 的CaCl2,对通风发酵过程中的不同K^＋、Ca^2＋浓度影响啤酒酵母代谢产6种有机酸含量的动态变化进行了跟踪检测.研究结果表明,K^＋、Ca^2＋可能通过作用于酵母细胞膜上的膜蛋白或调控生理代谢网络中代谢流相关的酶,从而使不同的K^＋、Ca^2＋浓度影响啤酒酵母响应产酸的峰值和峰值响应时间;在通风发酵过程中,啤酒酵母代谢产乳酸较多（多达8.3 mg/mL）,产琥珀酸较少（不超过250 μg/mL）;发酵终点时,随K^＋、Ca^2＋浓度增大,啤酒酵母代谢产酒石酸和琥珀酸等含量减少.     

非酿酒酵母对有机酸类碳源代谢特征的研究

以酿酒酵母（Saccharomyces cerevisiae）RV002为对照,将发酵毕赤酵母（Pichia fermentans）JT-1-3和季也蒙毕赤酵母（Meyerozyma guilliermondii）JP-4-2分别接种于以L-苹果酸、柠檬酸、葡萄糖和乙醇为单一碳源或双碳源的培养基中,测定酵母生物量及碳源代谢动态变化,研究两株非酿酒酵母（non-Saccharomyces）对碳源尤其是有机酸类碳源的利用情况。结果表明,发酵毕赤酵母JT-1-3对L-苹果酸的代谢率最高（49.74%）,季也蒙毕赤酵母JP-4-2对柠檬酸的代谢率最高（28.30%）;除发酵毕赤酵母JT-1-3在L-苹果酸与乙醇共存的培养基中L-苹果酸消耗量（58.88%）增加外,其余情况下,乙醇或葡萄糖的存在会抑制非酿酒酵母对L-苹果酸或柠檬酸的利用,这将为进一步利用两株非酿酒酵母酿造酸度适宜的果酒提供理论基础。     

发酵条件对梨醋酿造过程中有机酸的影响

采用反相高效液相色谱分析了梨酒、梨醋中有机酸,研究了发酵条件对其有机酸的影响。研究结果表明:酿酒酵母在酒精发酵过程中可代谢梨汁中有机酸。与梨汁相比,梨酒中的草酸、奎宁酸含量下降,乳酸、琥珀酸含量上升。梨酒中乙酸、琥珀酸随着酒精发酵温度的升高而增加,而苹果酸、乳酸、奎宁酸则随着酒精发酵温度的升高而降低。醋酸菌具有代谢各种有机酸的功能,在醋酸发酵过程中均消耗柠檬酸,而产生和积累酒石酸。梨醋中酒石酸、奎宁酸、莽草酸、琥珀酸和乳酸随着醋酸发酵温度的升高而增加,苹果酸和草酸几乎不受发酵温度的影响。发酵方式影响醋酸菌对有机酸的代谢。     

啤酒发酵过程有机酸代谢特征变化规律的研究

对发酵过程中几种主要有机酸的变化进行了初步研究。研究结果表明,有机酸大致可以分为三类：第I类,发酵过程中基本不分泌到胞外、或被酵母吸收的有机酸,包括柠檬酸、苹果酸、富马酸和甲酸;第II类,发酵过程中有较多增加的有机酸,包括乳酸和乙酸;第III类,麦汁中含量较少,但发酵过程中会大量产生,包括的有机酸为琥珀酸。     

耐温絮凝酵母Saccharomyces cerevisiae KF—7的细胞活性及发？…

本文通过不同温度下对絮凝酵母Ｓａｃｃｈａｒｏｍｙｃｅｓ ｃｅｒｅｖｉｓｉａｅ ＫＦ－７胞内储存糖含量、有机酸综合值ＯＳＲ及发酵性能的分析研究，探讨了絮凝酵母的耐温性及细胞活性。研究结果表明：酵母细胞的耐温性及细胞活性与细胞内的海藻糖含量密切相关；酵母分泌的有机酸综合值ＯＳＲ可以直观反映酵母细胞活性；在一定温度范围内，低温有利于酵母增殖，高温有利于酒精发酵；提高发酵温度可以提高酵母活性，增加出酒率。     

耐温絮凝酵母Saccharomyces cerevisiae KF-7的细胞活性及发酵特性研究

本文通过不同温度下对絮凝酵母ＳａｃｃｈａｒｏｍｙｃｅｓｃｅｒｅｖｉｓｉａｅＫＦ－７胞内储存糖含量、有机酸综合值ＯＳＲ及发酵性能的分析研究，探讨了絮凝酵母的耐温性及细胞活性。研究结果表明：酵母细胞的耐温性及细胞活性与细胞内的海藻糖含量密切相关；酵母分泌的有机酸综合值ＯＳＲ可以直观反映酵母细胞活性；在一定温度范围内，低温有利于酵母增殖，高温有利于酒精发酵；提高发酵温度可以提高酵母活性，增加出酒率。发酵试验结果表明絮凝酵母ＫＦ－７为一株性能优良菌株。     

生物工程技术在苹果酸-乳酸发酵中的应用

苹果酸－乳酸酵母是葡萄酒中的苹果酸在乳酸菌的作用下转变为乳酸和CO2的过程，它有利于葡萄酒质量的提高。介绍了生物反应器和采用基因工程方法构建的苹果酸－乳酸酵母进行葡萄酒生物降酸的技术。（1）游离生物反应器。使用细胞循环式发酵以在到高细胞生物量；或控制连续发酵；（2）固定化细胞，有包埋法和吸附法。（3）固定化酶技术。（4）苹果酸－乳酸酵母的应用。构建的苹果酸－乳酸酵母能在7d内将苹果酸转为乳酸，并具有良好的酿酒特性。（津京）     

顺序和同时接种乳酸菌对马瑟兰葡萄酒品质的影响

为探讨乳酸菌、非酿酒酵母和酿酒酵母对马瑟兰葡萄酒风味品质的影响，以戴尔有孢圆酵母、酿酒酵母与乳酸菌顺序接种（sequential inoculation，SEQ）和同时接种（simultaneous inoculation，SIM）为试验处理，以不接种乳酸菌自发进行苹果酸-乳酸发酵为对照（control，CK）酿造马瑟兰葡萄酒。采用顶空固相微萃取结合气相色谱质谱技术测定挥发性香气成分、高效液相色谱测定有机酸含量，并对葡萄酒进行感官评价。试验结果表明：SIM 处理同CK 处理的葡萄酒颜色差异明显；SIM 处理缩短了苹果酸-乳酸发酵进程，且葡萄酒柠檬酸和酒石酸含量显著低于SEQ 处理和CK处理（P＜0.05）；SEQ 和SIM 处理显著提高了葡萄酒中酯类物质含量（P＜0.05），尤其是己酸乙酯和己酸异戊酯等化合物含量，CK 处理异戊醇显著高于其它处理；主成分分析表明，SIM 处理可以改变葡萄酒的香气轮廓，提升葡萄酒的复杂性，感官评分最高。     

小麦啤酒有机酸组成的优化研究

分别考察了酵母种类和小麦芽品种对成品小麦啤酒有机酸组成的影响.基于德国品牌小麦啤酒的有机酸组成,确定了小麦啤酒主要有机酸的较佳组成范围,并且通过L_9(3~3)正交实验设计,最终确定了较佳的发酵工艺参数:麦汁pH5.5;接种量为1.0×10~7个/mL;主酵温度10℃.研究表明,选择产酸合理的酵母菌株和优良的小麦芽以及使用较佳发酵工艺,可以有效地降低乙酸、琥珀酸含量和增加柠檬酸含量,进而优化小麦啤酒的有机酸组成,这对改善小麦啤酒的酸感和风味都有很大的指导意义.     

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     

打瓜啤酒生产工艺的优化

在啤酒生产过程中添加打瓜,通过单因素和正交试验对打瓜啤酒酿造工艺进行优化,利用电子舌、气相色谱（GC）和高效液相色谱（HPLC）测定打瓜啤酒味觉特征、香味物质和有机酸。结果表明,打瓜啤酒最佳生产工艺条件为打瓜汁添加量18%,酵母添加量为1.0 g/L、初始pH值5.2、主发酵温度10.5 ℃。在此优化条件下,打瓜啤酒的感官评分为93.3分。打瓜啤酒丰富度饱满,整体滋味偏咸味和鲜味;通过GC在打瓜啤酒中检测出10种香气成分,其中醛类物质1种、醇类物质4种、酯类物质5种;HPLC检测分析的9种有机酸均有检出,其中柠檬酸含量最高,为（9.30±0.038）g/L,是打瓜啤酒有机酸组成的主体。     

THE CONTENT OF SOME ORGANIC ACIDS IN BEER AND OTHER FERMENTED MEDIA

The acid content of a range of ales and lagers has been measured for some organic acids related to the Krebs cycle, and found to vary widely. Acetate, pyruvate, lactate, succinate, pyroglutamate, malate and citrate were present in all cases and α-ketoglutarate was usually detected. α-Hydroxyglutarate was recognized in a number of beers. The effect of the acids on the pH of beer is assessed. The strain of yeast which is used markedly influences the levels of all acids except pyroglutamate and the conditions of yeast propagation have a substantial influence on the extent of acid accumulation. During the fermentation of wort and synthetic media the extent of organic acid excretion is proportional to the extent of fermentation, but the nature of the acids which are excreted varies during the fermentation period. In synthetic media, nitrogen source is shown to have a substantial effect on the accumulation of organic acid. Pyruvate and acetate levels vary inversely towards the end of fermentation, suggesting that yeast converts pyruvate to acetate.     

THE BIOCHEMISTRY OF MATURATION

Negative and positive aspects of maturation are respectively related to aroma and taste modifications. Vicinal diketones, hydrogen sulphide, acetyldehyde being primarily responsible for ‘green’ beer flavours an important feature of maturation is the adjustment of their concentration during the lagering period. The role of secondary fermentation in the removal of these undesirable by-products and the importance of sulphury compounds in determining the typical character of lager beer are reviewed. Particular emphasis is placed upon new enzymatic and genetic approaches to overcome vicinal diketone problems in accelerated fermentation systems using free and immobilized cells. The presence in beer of amino acids, peptides, nucleotides and organic as well as inorganic phosphates is, in part, due to the secretion of these materials by the yeast during lagering. Most of these compounds are contained in internal pools and their actual participation in flavour maturation depends upon intracellular breakdown and accumulation, changes in cell permeability and subsequent exchange possibilities between the yeast cell and the surrounding beer. Participation and practical implications of medium chain length fatty acids in the development of autolytic and yeasty flavours are discussed.     

啤酒高浓酿造中氨基酸对酵母发酵性能和啤酒色值的影响

探讨在24?°P高浓啤酒发酵过程中8?种氨基酸（Met、Phe、Trp、Arg、His、Ile、Leu、Lys）的不同添加量（分别为原麦汁中相应氨基酸含量的0.5、1?倍和2?倍）对酵母生理特性、发酵性能和啤酒色值的影响。结果表明：8?种氨基酸的补充可显著提高麦汁发酵度、乙醇产量,促进酵母生长,提高酵母活细胞率,改善啤酒色值。其中,补充1?倍氨基酸的高浓麦汁发酵性能较好,与对照组相比,发酵度、乙醇产量、最大悬浮酵母细胞数和发酵结束时的酵母活细胞率分别提高了6%、17%、11%和10%。添加氨基酸的高浓酿造啤酒经稀释后,啤酒色泽依然鲜亮,且添加1?倍氨基酸酿造而成的啤酒经稀释后色差（ΔE）最小,色泽最接近青岛纯生啤酒。     

啤酒发酵过程中酸类物质的变化与酵母活力关系的研究

研究了发酵过程中部分有机酸、脂肪酸含量的变化趋势与酵母活力的关系，以及使用不同代数的酵母对发酵液有机酸和脂肪酸含量、组成的影响；并且对比了不同使用代数酵母泥离心液和发酵液相应有机酸、脂肪酸含量的差值，以及差值与酵母活力的关系。     

THE ORIGIN OF THE MEDIUM CHAIN LENGTH FATTY ACIDS PRESENT IN BEER

The medium chain length fatty acids that are excreted during fermentation are produced by synthesis and not by degradation. The fermentation of a wort supplemented with propionic acid (C₃) or valeric acid (C₅) leads to the excretion of nonanoic acid (C₉) in addition to the usual even chain acids. C₉ acid was not detected in the beer when the inoculated yeasts contained a high proportion of pentadecanoic acid (C₁₆) and heptadecanoic acid (C₁₇) or when the C₁₇ acid was added to the wort, demonstrating that a degradative route is unimportant. The content of the medium chain length fatty acids in beer varies directly with their content in yeast; thus the fatty acid composition of the beer reflects changes in the content of these acids in yeast brought about by alteration in the supply of oxygen or by the addition of C3 acid to wort.