2种葡萄酒酵母发酵黑莓酒的降酸效果研究

研究采用反相高效液相色谱法测定两种酵母发酵的黑莓酒中主要有机酸的种类及含量,为筛选出适合黑莓酒同步发酵降酸的酵母提供依据.对61A和71A酵母发酵的黑莓酒进行RP-HPLC分析,结果表明,2种酵母发酵的黑莓酒中主要有机酸都是苹果酸,其次是酒石酸、醋酸和乳酸.与61A酵母相比,用71A酵母发酵的黑莓酒中苹果酸、酒石酸、醋酸、乳酸、草酸和琥珀酸分别下降了3.08mg/mL、0.6 mg/mL、0.64 mg/mL,、0.69mg/mL、0.3 mg/mL和0.88mg/mL,柠檬酸含量上升为0.42mg/mL,总酸降低了5.77 mg/mL,说明71A酵母在酒精发酵的同时具有较好的降酸效果,足酿造黑莓酒的良好菌株.     

Ethyl carbamate production by selected yeasts and lactic acid bacteria in red wine

This study was designed to investigate the formation of ethyl carbamate during the fermentation of musts of Vitis vinifera L. cv. Tempranillo and Cabernet Sauvignon by selected yeasts in different conditions of temperature and pH. Secondary malolactic fermentation was then induced by selected lactic acid bacteria. The results indicate an increased concentration of ethyl carbamate after malolactic fermentation, irrespective of the bacteria used or the prevailing physicochemical conditions.     

Survey of hydrogen sulphide production by wine yeasts

Twenty-one strains of commercial wine yeasts and 17 non-Saccharomyces species of different provenance were surveyed for their ability to produce hydrogen sulphide in synthetic grape juice medium indicator agar with different nitrogen sources, as well as in natural grape juice. Bacto Biggy agar, a commercially available bismuth-containing agar, was used to compare our results with others previously reported in the literature. Under identical physiological conditions, the strains used in this study displayed similar growth patterns but varied in colony color intensity in all media, suggesting significant differences in sulphite reductase activity. Sulphite reductase activity was absent for only one strain of Saccharomyces cerevisiae. All other strains produced an off-odor to different extents, depending significantly (P <0.05) on medium composition. Within the same species of some non-Saccharomyces yeasts, strain variation existed as it did for Saccharomyces. In natural musts, strains fell into three major groups: (i) nonproducers, (ii) must-composition-dependent producers, and (iii) invariable producers. In synthetic media, the formation of sulphide by strains of S. cerevisiae results from the reduction of sulphate. Therefore, this rapid screening methodology promises to be a very useful tool for winemakers for determining the risk of hydrogen sulphide formation by a given yeast strain in a specific grape juice.     

Differentiation between fermenting and spoilage yeasts in wine by total free fatty acid analysis

Differentiation between fermenting and spoilage yeasts in wine was estimated by cellular fatty acid profiles. Forty-two strains of yeasts representing 17 genera were grown on a defined liquid medium for 48 h at 25°C in a rotary shaker. After saponification of yeast cells, free fatty acid extracts were analysed by gas chromatography. Multivariate analysis was performed by Principal Components Analysis (PCA) to define clusters of fatty acids and yeasts. Strains were characterised especially by long-chain fatty acids, palmitic (C16) to linolenic (C18:3) acid under aerobic culture. Nevertheless, most of Saccharomyces cerevisiae and also Dekkera bruxellensis (former names D intermedia and Brettanomyces lumbicus) synthesized medium-chain fatty acids, octanoic (C8) acid to dodecanoic (C12) acid. With this method it was possible to differentiate fermenting grape yeasts such as S cerevisiae from spoilage yeasts on the basis of the absence of linoleic (C18:2) and linolenic (C18:3) acids. However, the method seemed unreliable for the identification of strains, more particularly those of species of S cerevisiae.     

Metabolites produced during fermentation of wine by mixed cultures of yeasts and lactic acid bacteria

Certain aspects of the interrelationship between associations of yeasts and lactic acid bacteria during the fermentation of grape must were studied. Action bySaccharomyces pastorianus gave rise to increased formation of keto acids and residual metabolites. Addition ofLactobacillus plantarum to the fermentation process may inhibit alcoholic fermentation and considerably increased the production of acetaldehyde andl(+) andd(–)lactic acids.

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Entstehung von Metaboliten bei der Fermentation von Wein durch Mischkulturen von Hefen und Milchsäurebakterien

Zusammenfassung Es wurden Wechselbeziehungen zwischen Hefen und Milchsäurebakterien während der Fermentation von Traubensaft untersucht.Saccharomyces pastorianus bewirkte ein Ansteigen der Ketosäurenproduktion und der Bildung von Rest-Metaboliten. Das Hinzufügen vonLactobacillus plantarum beim Gärungsprozess kann die alkoholische Gärung unterdrücken und hat die Produktion von Acetaldehyd sowie vonl(+)- undd(–)-Milchsäuren beträchtlich gesteigert.

     

L-苹果酸的开发与应用

介绍了L-苹果酸的性能、生产的主要技术路线与最佳的操作条件及有关进展情况。阐述了国内外研究开发的现状与发展趋势．并探讨了扩大应用范围等的前景与市场需求。     

Selection and modification of yeasts and lactic acid bacteria for wine fermentation

Historically, the fermentation of grape juice to wine has been carried out by indigenous yeasts found on the berry. However, in newer wine regions, e.g. the USA, inoculation with selected wine yeast strains is employed. Grape juice is high in nutritional factors and difficulties in fermentation usually arise from the inhibitory effects of the high concentration of sugar initially present and the ethanol produced. A secondary fermentation, brought about by indigenous or added lactic acid bacteria, converts malic acid to lactic acid and carbon dioxide and often occurs. This ‘malolactic’ fermentation is usually slow. For both yeast and bacterial fermentations strain selection is based more on fermentation performance than on sensory characteristics of the wine, with increased tolerance of the yeast to ethanol and of the bacteria to low pH being emphasized. Attempts to increase the malolactic fermentation rate have been made by cloning and transferring the malolactic gene from Lactobacillus to wine yeast. In early attempts to produce wines with enhanced or novel sensory characteristics a leucine-less mutant of a homothallic wine yeast has been obtained which does not produce isoamyl alcohol.     

Formation of α-ketoglutaric acid by wine yeasts and its oenological significance

α-Ketoglutaric acid was measured enzymically in wines made in the laboratory from three grape varieties by pure cultures of 12 wine yeasts of the genus Saccharomyces. The results were confirmed with the same juices and 4 yeasts on pilot-plant scale in replicated 30 gallon lots. Mean values for the 12 yeasts ranged from 9 to 117 ppm (overall mean 53). In any one juice the yeasts differed by at least 10-fold in the amounts produced, and certain yeasts produced consistently high or low yields in all juices. The amounts of α-ketoglutaric acid produced depended somewhat on the grape juices used, even though these had comparable pH values, and a significant yeast-juice interaction occurred. The amount of α-ketoglutaric acid formed during fermentation at 15° was 60 per cent of that formed at 25°, and over twice as much was formed at pH 4.2 as at pH 3.0, using four yeast strains. Formation of α-ketoglutaric and pyruvic acids were not significantly correlated. The α-ketoglutaric acid content of 18 white table wines made under comparable conditions on pilot-plant scale from different grape varieties using the same yeast strain ranged from 38 to 152 ppm (mean 90). The significance of the results is discussed, particularly in relation to the binding of sulphur dioxide in wine, and recommendations are given on how to make wines which are low in α-ketoglutaric acid. Formation of α-ketoglutaric acid by three yeasts in a chemically defined medium was lower with increased amounts of nitrogen as ammonium sulphate and higher in the presence of L-glutamic acid, both being used separately as sole nitrogen sources. These findings are discussed in relation to the rǒle of α-ketoglutaric acid in nitrogen metabolism of yeasts.     

Influence of different yeasts on the growth of lactic acid bacteria in wine

The influence of various yeasts on the growth of lactic acid bacteria in wine was tested by inoculating Lactobacillus hilgardii, L. brevis and two strains of Leuconostoc mesenteroides into experimental wines made with twelve different yeasts of the genus Saccharomyces. Wines made from juice which had been infected with several spoilage yeasts and then fermented with a wine yeast were also tested in this way. It was found that the yeasts differed considerably in their effects on bacterial growth. In some of the experimental wines bacterial growth was delayed or failed altogether. Generally, the unfavourable influence of any yeast on bacterial growth was much reduced if the wines were left in contact with the yeast cells for some weeks after the fermentation. The significance of these results in relation to the occurrence of malo-lactic fermentation in commercial wineries is discussed.     

3株哈密瓜酒酵母的分子生物学鉴定

通过对从哈密瓜酒生产中分离获得的3株哈密瓜酒酵母进行发酵性能分析,发现其中1株发酵异常.采用26S rDNA D1/D2区序列分析法对酵母进行分子鉴定,经序列比对及构建系统发育树分析,得出1号和3号酵母为酿酒酵母,与酿酒酵母模式菌株序列相似性在99％以上;2号酵母鉴定为高里毕赤酵母,与高里毕赤酵母(Pichia guilliermondii)相似性为99％.进一步对3株酵母的同源序列与其他种之间的发育关系进行了分析,发现有很好的相似度,表明26S rDNA D1/D2区域序列分析方法能够应用于酿酒酵母的分子生物学鉴定,并可以作为生产中酵母污染检测的工具.     

3种不同酵母产蓝莓酒风味物质的研究

利用3种不同酵母（T1、FK3-2、LM6）发酵生产蓝莓酒,并对其主发酵期的风味物质进行研究。气相色谱（GC）检测发现,酵母LM6发酵的蓝莓酒后期异丁醇、异戊醇和丁酸乙酯含量较高,分别为2.38 mg/mL、2.13 mg/mL、2.05 mg/mL。高效液相色谱（HPLC）检测发现,菌株LM6发酵的蓝莓酒在28 d时有机酸种类减少,草酸含量增加至3.17 mg/mL,L-苹果酸含量降至0.09 mg/mL、乳酸含量降至9.87 mg/mL,琥珀酸含量降至3.06 mg/mL。电子舌检测中发现FK3-2和T1酵母菌产生的酸味明显且所产风味相似;LM6酵母菌产生的风味较均衡,但鲜味、咸味及丰富性更明显。综合比较发现,LM6酵母菌更适合发酵蓝莓酒。     

Spoilage yeasts in the wine industry

Yeasts play a central role in the spoilage of foods and beverages, mainly those with high acidity and reduced water activity (a(w)). A few species are capable of spoiling foods produced according to good manufacturing practices (GMPs). These can survive and grow under stress conditions where other microorganisms are not competitive. However, many of the aspects determining yeast spoilage have yet to be clarified. This critical review uses the wine industry as a case study where serious microbiological problems are caused by yeasts. First, the limitations of the available tools to assess the presence of spoilage yeasts in foods are discussed. Next, yeasts and factors promoting their colonisation in grapes and wines are discussed from the ecological perspective, demonstrating that a deeper knowledge of vineyard and winery ecosystems is essential to establish the origin of wine spoilage yeasts, their routes of contamination, critical points of yeast infection, and of course, their control. Further, zymological indicators are discussed as important tools to assess the microbiological quality of wines, although they are rarely used by the wine industry.The concepts of the susceptibility of wine to spoilage yeasts and wine stability are addressed based on scientific knowledge and industrial practices for monitoring yeast contamination. A discussion on acceptable levels of yeasts and microbiological criteria in the wine industry is supported by data obtained from wineries, wholesalers, and the scientific literature.Finally, future directions for applied research are proposed, involving collaboration between scientists and industry to improve the quality of wine and methods for monitoring the presence of yeast.     

鸭梨酒酵母菌的筛选

从鸭梨、苹果、枣、葡萄等水果的果皮中筛选出了一株适于鸭梨酒酿造的酵母菌 ,命名为GY。其发酵酒的酒精含量符合低度果酒 (7%V/V～ 12 %V/V)的要求 (由于鸭梨本身含糖量低 ,在本研究中鸭梨酒的最终酒精含量要求达到 7%V/V～ 8%V/V即可 ) ,此酵母菌能耐受 16 % (V/V)的酒精度和 375× 10 -6的二氧化硫浓度。另外 ,该酵母菌具有较好的耐糖性能 ,在 70 %的糖液中仍能起酵。     

Studies on acetate ester production by non-saccharomyces wine yeasts.

A double coupling bioreactor system was used to fast screen yeast strains for the production of acetate esters. Eleven yeast strains were used belonging to the genera Candida, Hanseniaspora, Metschnikowia, Pichia, Schizosaccharomyces and Zygosacharomyces, mainly isolated from grapes and wine, and two wine Saccharomyces cerevisiae strains. The acetate ester forming activities of yeast strains belonging to the genera Hanseniaspora (Hanseniaspora guilliermondii and H. uvarum) and Pichia (Pichia anomala) showed different substrate specificities and were able to produce ethyl acetate, geranyl acetate, isoamyl acetate and 2-phenylethyl acetate. The influence of aeration culture conditions on the formation of acetate esters by non-Saccharomyces wine yeast and S. cerevisiae was examined by growing the yeasts on synthetic microbiological medium. S. cerevisiae produced low levels of acetate esters when the cells were cultured under highly aeration conditions, while, under the same conditions, H. guilliermondii 11104 and P. anomala 10590 were found to be strong producers of 2-phenylethyl acetate and isoamyl acetate, respectively.     

不同菌种对青稞酒发酵的影响

以青稞酒传统生产工艺为基础,研究了发酵过程中使用不同菌种对青稞酒感官品质和出酒率的影响。结果表明,以传统的青稞整粒固态发酵工艺,以单一菌种发酵生产的青稞酒产品其出酒率和产品感官品质几乎没有提高,而使用酿酒酵母和甜酒曲复合菌种,可以在保证产品观感质量的前提下相对于当地青稞酒生产工艺提高出酒率1倍以上。      

葡萄酒酵母选育技术研究进展

文中主要从酿酒酵母及非酿酒酵母的选育展开论述,概括介绍了目前葡萄酒酵母的选育技术及成果,同时根据文献对国内和国外的研究现状作了分析,并对今后的研究趋势做了推测.     

菠萝果酒酿造酵母菌的筛选研究

研究了4株酵母菌在菠萝汁中的生长特征以及发酵过程中的总糖、还原糖、总酸和酒精度等理化指标的变化规律,对菠萝酒进行感官评定,筛选出酿造菠萝果酒的优良菌株.结果表明,4号酵母菌生长繁殖快,发酵性能优良,能耐高酒精和高糖,该酵母菌在菠萝汁中发酵7d后总糖为11.524g/L,还原糖为4.471g/L,总酸为4.75g/L,pH值为4.64,酒精度为18.096%vol,感官评定得分94分.确定4号酵母菌(威尔)是菠萝果酒酿造的优良菌株.     

L-苹果酸一步发酵法的工艺研究及浅析

国内外有机酸发酵中,苹果酸发酵的方法分为两种,即"一步法"和"两步法".以米曲霉为菌种,通过一步发酵法生产L-苹果酸,应用正交实验设计法对温度、接种量、起始糖浓度以及碳酸钙加量4因素进行探索,确定最优工艺条件.结果表明,最适发酵条件为温度34.0℃、起始糖浓度为100.0 g/L、接种量8.0%、碳酸钙加量75.0 g/L.     

高效降解OTA的发酵工业用菌株筛选及鉴定

为筛选一株产L-苹果酸能力强的黑曲霉菌株,通过紫外诱变与高浓度放线菌酮迭代诱变的方法,将野生型菌株诱变为一株产L-苹果酸能力强的黑曲霉菌株,并对其种子或发酵培养基成分进行优化。结果表明,诱变所得一株产L-苹果酸能力强的黑曲霉CGMCC NO. 40550菌株,其摇瓶发酵时种子培养基最适葡萄糖浓度为60 g/L、最适氮源为（NH₄）₂SO₄ 4.95 g/L,黑曲霉菌球形成最优转速220 r/min;发酵培养基最适葡萄糖和最适（NH₄）₂SO₄浓度分别为180 g/L和4.95 g/L,CuSO₄·5H₂O为其生产苹果酸最佳微量元素,最适添加量为 0.065 g/L。通过单因素实验的优化,优化后的培养基更有利于L-苹果酸的合成,发酵96 h时L-苹果酸产酸量达到18.15 g/L,显著提高了245%,L-苹果酸占总酸的百分比达到71.69%。本研究为L-苹果酸的工业化生产奠定基础。

     

黄曲霉积累L-苹果酸代谢机制初探

L-苹果酸是生物体内三羧酸循环的成员之一,在食品、医药、日用化工等部门具有广泛的用途.文中从限氧发酵、碳酸钙的添加量、乙醛酸循环和TCA循环相应酶的抑制剂几个方面初步探讨黄曲霉积累L-苹果酸的代谢机制,得出CO2固定途径是积累L-苹果酸的主要途径.