Physiological features of Schizosaccharomyces pombe of interest in making of white wines

This work studies the physiology of Schizosaccharomyces pombe strain 938 in the production of white wine with high malic acid levels as the sole fermentative yeast, as well as in mixed and sequential fermentations with Saccharomyces cerevisiae Cru Blanc. The induction of controlled maloalcoholic fermentation through the use of Schizosaccharomyces spp. is now being viewed with much interest. The acetic, malic and pyruvic acid concentrations, relative density and pH of the musts were measured over the entire fermentation period. In all fermentations in which Schizo. pombe 938 was involved, nearly all the malic acid was consumed and moderate acetic concentrations produced. The urea content and alcohol level of these wines were notably lower than in those made with Sacch. cerevisiae Cru Blanc alone. The pyruvic acid concentration was significantly higher in Schizo. pombe fermentations. The sensorial properties of the different final wines varied widely.     

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 yeast strain on binding of sulphur dioxide in wines,and on its formation during fermentation

The amounts of sulphur dioxide bound by acetaldehyde, pyruvic acid and α-ketoglutaric acid during fermentation of three grape juices by eight wine yeasts (Saccharomyces sp.) are reported. These constituents accounted for 49–83 % (mean 69) of the measured bound SO₂, depending on the yeast strain and juice. the maximum range of concentrations of the binding components for individual wines were 10–48 ppm for acetaldehyde, 9–77 ppm for pyruvic acid and 5–63 ppm for α-ketoglutaric acid, depending on yeast strain and grape juice. the validity of the calculations was verified by an experiment with SO₂ and the three binding compounds in a multicomponent model system. The acetaldehyde content was related to the total SO₂ present, which itself was determined by the strain of yeast. SOz bound in the wines after a further SO₂ addition was correlated significantly with pyruvic and α-ketoglutaric acids, but not with acetaldehyde. Certain yeasts produced SO₂ during fermentation in grape juice and in synthetic media with defined sulphur sources. More SO₂ was produced at pH 3.6 than 3.0 in the absence of added sulphate in grape juice. Sulphate was the best sulphur source for SO₂ production in synthetic media, although some yeasts were able to produce smaller amounts of SO₂ from l-cysteine and reduced glutathione.     

昌黎产区产酶酵母多样性及其应用潜力分析

利用优选酵母混菌发酵,是改善葡萄酒品质及增香的有效途径。以赖氨酸培养基、WL培养基和产酶筛选培养基为基础,分析了昌黎4个葡萄园土样中产酶酵母的多样性,结果表明,该地区产酶酵母丰富,50%的产酶酵母能同时产2种或2种以上的酶。对产酶酵母酿造因子耐性分析表明,菌株GY1、M9和J24具有高糖耐性,适合于高糖低醇果酒酿造;菌株GY13和M9具有高酒精耐性,酒精含量为15.000%时活性较强;菌株J24和GY1具有高酸耐性,pH 2.5时具有较高活性;7株菌15℃时具有较高活性。综合分析发现,菌株M9(Torulaspora delbrueckii)同时产β-葡萄糖苷酶、果胶酶和纤维素酶,除酸耐性较差外,具有高糖、高酒精度和低温耐性,在高糖低醇果酒酿造中具有很大应用潜力。     

Use of unripe grapes harvested during cluster thinning as a method for reducing alcohol content and pH of wine

Background and Aims: Deep red full‐bodied wines can only be obtained from grapes with complete phenolic maturity, which frequently produce wines with high pH and alcohol content. The present study focuses on a new procedure for simultaneously reducing pH and ethanol content. Methods and Results: Grapes from cluster thinning were used to produce a very acidic low‐alcohol wine. The wine was treated with high doses of charcoal and bentonite. This odourless and colourless wine was used to reduce pH and ethanol content of wine produced from grapes, which had reached complete phenolic maturity. The anthocyanin and proanthocyanidin concentrations, the mean degree of polymerisation and the monomeric composition of proanthocyanidin of reduced‐alcohol wines were similar to those of their corresponding controls. Since the pH was lower, the colour of the reduced‐alcohol wines was more intense. No significant differences were found between reduced‐alcohol wines and their controls by triangle sensory tests using dark glasses for two of the three studied cultivars. Conclusion: The procedure described allowed production of wines with reduced alcohol content and pH, while retaining similar phenolic content and sensory properties. Significance of the Study: The proposed procedure is easy to apply, does not require specific equipment and offers a means of addressing the problem of wines developing high ethanol and low pH as a result of over‐ripening of grapes.     

Malolactic Fermentation and Secondary Metabolite Production by Oenoccocus oeni Strains in Low pH Wines

Abstract: Different wine varieties, including some with low pH, were studied to determine the ability to grow and produce secondary metabolites of a previously selected autochthonous Oenococcus oeni strain (C22L9), compared with a commercial strain. Monitoring of malolactic fermentation (MLF) was carried out by microbiological and chemical analysis of wines. The concentration of some major volatile compounds and biogenic amines in wines before and after malolactic fermentation was also determined. The results showed major differences in MLF duration both between wines and strains, although the differences between strains were slight for most of the analyzed compounds. Statistically significant differences in citric acid degradation were found in all wine varieties and it was confirmed that O. oeni C22L9 is a poor degrader of citric acid; this means that MLF can be prolonged without the risk of producing high concentrations of diacetyl and acetoin. Sensory analysis of wines after MLF showed similar characteristics in wines from both strains. This study thus shows that O. oeni C22L9 possesses even better sensory and fermentation properties than the commercial strain and can be used in wines with different characteristics, which makes it highly valuable for industrial use. Practical Application: The increasingly use of grape varieties of low pH in winemaking and the higher alcohol content of wines, as a consequence of the climatic change, make interesting the study of the behavior during MLF of O. oeni strains in order to determine their ability to grow, when growth conditions are not optimal, and to produce secondary metabolites. A comparative study was conducted using an autochthonous O. oeni strain (C22L9) and a commercial O. oeni strain and 4 wine varieties.     

Formation of vitisin A during red wine fermentation and maturation

During alcoholic fermentation the formation of vitisin A occurred mainly in the period between 20% and 85% glucose utilisation, when the concentrations of the precursors, malvidin-3-glucoside and pyruvic acid were at a maximum. The maximum rate of vitisin A synthesis of 11 mg/Lh occurred when 57% of glucose had been utilised and 440 mg/L malvidin-3-glucoside, 114 mg/L pyruvic acid and 3.1 mg/L measured oxygen were present. During maturation, production of vitisin A appeared to be linked with the availability of a suitable oxidant. Synthesis of vitisin A continued for approximately 6 months of maturation in air-tight bottles while the precursors were available. In wines that underwent malolactic fermentation, the malolactic bacteria consumed pyruvic acid and thereby limited the production of vitisin A. At the end of a 12 month maturation period, the concentration of vitisin A declined in wines made without any malolactic fermentation, even though malvidin-3-glucoside and pyruvic acid were still present. This indicated that there was a shortage of a suitable oxidant required to complete the reaction.     

Assessing the anthocyanic composition of Port wines and musts and their free radical scavenging capacity

The anthocyanic composition of must and different types of Port wine (Ruby, LBV and Tawny from nine producers) was evaluated by HPLC. In addition, the radical scavenging activity of these Port wines was evaluated by using DPPH radical microassay. Musts, Ruby and LBV Port wines presented similar qualitative profiles of anthocyanins and quantitative differences, ranging from 382-601, 92-156 and 32-78 mg of malvidin-3-O-glucoside equivalent/l, respectively. Anthocyanins in Tawny Port wines ranged between not detected and 51 mg of malvidin-3-O-glucoside equivalent/l. Principal component analysis was applied to the percentage (in weight) of each compound in relation with the total content of anthocyanins. PCA yielded two components explaining 90.9% of the total variance. The first PC correlates positively with the glucoside derivatives of delphinidin, cyanidin, petunidin and malvidin. The second PC correlates positively with pyruvic acid adducts of anthocyanins that resulted from vinification process. Ruby and LBV presented low amounts of glucoside derivatives and variable amounts of pyruvic acid adducts of anthocyanins. Acid hydrolysis allowed the identification of five anthocyanidins. Malvidin was the most abundant one. LBV Port wine showed the strongest antiradical activity, while Tawny Port wines revealed the lowest ones, with these wines presenting lower amounts of anthocyanic compounds owing to their “oxidative” aging in wooden barrels. These results suggest that aging process is an important factor influencing the antioxidant activity of Port wines.     

Indian Grape Wines: A Potential Source of Phenols,Polyphenols, and Antioxidants

Indian grape wines are analyzed for total phenolic content and antioxidant activity along with other parameters, such as pH, alcohol content, and reducing sugars. Concentration of polyphenols, like tannic acid, catechol, vanillin, caeffic acid, ferullic acid, and resveratrol, was quantified using reverse phase-high performance liquid chromatography and ultra high performance liquid chromatography. The red wines showed the highest concentration of phenolic content (6.5 ± 0.1 mg/ml) and antioxidant activity (84.60 ± 1%) as compared to white and port wines, while red wine R2 showed the highest radical scavenging activity among red wines and R4 showed the lowest total phenolic content. The white wine W3 showed less total phenolic content and antioxidant activity. Further, a positive correlation between phenolic content and antioxidant activity was observed.     

Use of grape racemes from Grillo cultivar to increase the acidity level of sparkling base wines produced with different Saccharomyces cerevisiae strains

The most important oenological characteristics of high-quality sparkling wines are aromatic aspect, taste persistence, perlage, high levels of acidity and low pH. Due to hot climate and reduced rainfall that characterize Sicily region, white grape varieties such as Grillo cultivar cultivated in this area are characterized by very low concentrations of malic and tartaric acids. Grillo cultivar is characterized by an intense production of raceme grapes with low pH and high content of tartaric and malic acids. These fruits possess the chemical properties useful to increase the amounts of acids in the final wines. With this in mind, the present research was carried out to test the ability of four Saccharomyces cerevisiae strains (CS182, GR1, MSE13 and MSE41) to ferment a raceme must with a pH of 2.9 at two concentrations (14° and 16° Babo degree) of total sugars. The inoculation of the strains was performed after a preadaptation at pH 2.5. The chemical parameters and kinetics of the fermentations were monitored. The experimental sparkling base wines were characterized by a very high total acidity with 16–17 g/L of tartaric acid and 9–10 g/L of malic acids. On the other hand, ethanol was detected at low values in the range 9–10% (v/v). The base wine obtained with GR1differed in their high acidity values, whereas trials inoculated with CS182 showed more intense odors and exotic fruit. Experimental wines produced in this study represent an innovative strategy for “blending wines” to produce sparkling wines in dry Mediterranean climate.     

Effect of six organic acids on staphylococcal growth and enterotoxin production.

Four Staphylococcus aureus strains were incubated at 37 degrees C for 24 h in broth progressively acidified with lactic, citric, ascorbic, acetic, pyruvic and propionic acids, and their survival rate and enterotoxin producing ability was studied. Acids were chosen based on their frequent use by the food industry. Periodically, samples were withdrawn to determine counts, pH and the presence of enterotoxins A, B, C, and D. For a given acid, the effect on growth and enterotoxin synthesis was different. The most inhibitory acid for the growth of strains FRI-100 and FRI-472 was pyruvic acid, for strain FRI-137 was lactic acid, all six acids were equally effective on strain S6. Lactic acid was very inhibitory to enterotoxin synthesis, but the effect on this parameter of acetic and citric acids was almost nil. Enterotoxins were seen to be inactivated at acid pH values; enterotoxin B was the most resistant to inactivation.     

A new chemically defined medium for wine lactic acid bacteria

Growth and activity of lactic acid bacteria is crucial for the production and quality of grape wines but studying their metabolism is difficult in wines or complex laboratory media because of the undefined substrate pools. This work presents a new chemically defined medium that meets the fastidious nutritional requirements of wine lactic acid bacteria and yields rapid and strong growth. The new medium is composed of 44 constituents and a precise protocol is provided for its preparation. Maximum specific growth rates and growth yields of the wine strains studied were comparable to those obtained in common laboratory media, and the new medium allows for various modifications, such as changing the medium pH to the wine range, addition of L-malic acid or utilization of different carbon sources while maintaining growth of wine lactic acid bacteria. The medium was successfully tested with 22 wine strains of the genera Oenococcus, Lactobacillus and Pediococcus. It is suggested that this chemically defined medium be considered for the investigation of the nutritional requirements and metabolism of wine lactic acid bacteria.     

Growth and Metabolism of L-malic Acid by Lactobacillus plantarum CECT 220 in a Defined Medium

Malic acid can be metabolized by lactic acid bacteria by two main pathways. Previous studies in malic acid degradation by malic and malolactic enzymes in wines and complex media have not been carried out in completely defined media. Some novel aspects of growth and metabolism of a L. plantarum strain in a defined medium were found. One of the most relevant results was that the strain could grow with malate as sole energy source added to the medium.     

Influence of different pH values and inoculation time on the growth and malolactic activity of a strain of Oenococcus oeni

The present study was undertaken to evaluate the influence of medium pH and inoculation time on the growth and malolactic activity of an Oenococcus oeni culture. Samples of a commercial white grape juice adjusted to pH 3.2, 3.4 or 3.6, and inoculated with Saccharomyces cerevisiae strain CY3079, were inoculated with a malolactic culture ( Oenococcus oeni strain 31) at the beginning, middle, and end of alcoholic fermentation. The results obtained from this single case study show that it is possible to inoculate the bacterial culture at the three different times during alcoholic fermentation without slowing down or stopping alcoholic fermentation or causing failure of MLF. However, pH and timing of bacterial inoculation were critical to how rapidly MLF starts. At pH 3.2 a lowering of bacterial viability was observed, but a more important reduction was recorded at all tested pH levels when the bacteria were inoculated halfway through alcoholic fermentation. When inoculation was carried out at the end of alcoholic fermentation, the presence of yeast seemed to favour bacterial viability and activity and bacteria performed MLF even in difficult conditions such as pH values around 3. In all wines malolactic fermentation was accompanied by total degradation of malic and citric acids and production of L-lactic acid, D-lactic and acetic acids.     

响应面法优化丙酮酸发酵培养基

采用响应面法(RSM)对丙酮酸发酵培养基成分葡萄糖、硫酸铵、蛋白胨进行优化,采用多元二次回归方程拟合3种因素与丙酮酸含量间的函数关系,并得到了最佳条件。在优化培养条件下,发酵液中丙酮酸的浓度由35.4g/L提高到41.57g/L,在5L罐的最佳浓度下丙酮酸产量71.23g/L比原产量65.76g/L提高8.3%。     

缬氨酸转氨酶拆分DL-缬氨酸的催化条件

利用具有缬氨酸转氨酶活性的工程菌对DL-缬氨酸进行拆分,考察了反应温度、pH值、底物摩尔比、底物浓度和金属离子对酶活性和底物转化率的影响。结果显示,该催化反应的最适反应条件为:反应温度是45℃,pH=9,L-缬氨酸与丙酮酸的摩尔比1∶8,DL-缬氨酸初始浓度为0.6 mol/L、丙酮酸初始浓度为2.4 mol/L,0.5 mmol/L的Mg2+和Na+对酶活性有明显的促进作用。     

高产核酸的热带假丝酵母诱变选育及其发酵条件优化

以热带假丝酵母（Candida tropicalis）HY4-17为出发菌株,采用电子束辐射技术诱变,经过初筛、复筛及遗传稳定性试验选育核酸高产菌株,通过Plackett-Burman试验、最陡爬坡试验及响应面法优化培养基,并优化接种量和发酵时间以提高核酸产量。结果表明,获得一株生长快速,遗传稳定的突变菌株EBI-21,其核酸含量较出发菌株HY4-17提高了25%。最佳培养基组成为：糖蜜185 g/L,硫酸铵3 g/L,硫酸锌0.05 g/L,硫酸镁0.5 g/L,硫酸亚铁0.05 g/L,磷酸1 mL/L,pH值为4.5。在优化培养基及接种量8%,发酵时间12 h条件下,核酸产量达到1.73 g/L,较未优化前提高了80.21%,培养时间缩短了10 h。     

不同酿酒酵母PDEs缺失突变株对磨盘柿果酒抗氧化及挥发性物质的影响

以磨盘柿子为原料,考察野生型及基因型为PDE1/Dpde1、Dpde1/Dpde1、PDE2/Dpde2和Dpde2/Dpde2的环核苷酸磷酸二酯酶基因缺失的酿酒酵母（Saccharomyces cerevisiae）发酵过程中可溶性固形物、总糖、乙醇体积分数和pH值等基本理化指标,总酚和总黄酮等活性物质以及2’-联氨-双-（3-乙基苯并噻唑啉-6-磺酸）二铵盐自由基、1,1-二苯基-2-三硝基苯肼自由基清除能力和总还原力等抗氧化能力的变化;并采用顶空固相微萃取技术与气相色谱-质谱联用相结合的方法,对不同菌株发酵的磨盘柿果酒挥发性物质成分进行比较分析。结果表明,在12?d达到发酵终点,4?株突变体发酵速率均比野生型快,其中Dpde2/Dpde2菌株的发酵速率最快;4?株突变体菌株酿造的磨盘柿果酒中总黄酮含量和抗氧化能力均高于野生型菌株,Dpde2/Dpde2菌株酿造的磨盘柿果酒抗氧化功效最强。PDEs基因缺失菌株与野生型酿酒酵母酿造的磨盘柿果酒中的风味物质种类差异不大,PDE2/Dpde2菌株酿造的磨盘柿果酒中酯类相对含量较高。     

突变型黄单胞杆菌产黄原胶条件的优化

该文对从甘蓝腐烂根部的土壤中分离选育的黄单胞杆菌C2发酵产黄原胶的条件进行了优化。通过单因素试验对培养基（碳源、氮源及碳氮源的浓度）和发酵条件（接种量、发酵时间、温度、pH、装液量）进行优化;为提高黄原胶的胶体性能,在培养中添加不同量的游离丙酮酸。结果表明,液态发酵生产黄原胶的最佳碳源、氮源分别为蔗糖55 g/L,鱼粉10 g/L。黄单胞杆菌C2菌株在接种量为4%,pH值为7,于28 ℃、130 r/min条件下发酵60 h时,黄原胶产量达到34 g/kg。添加游离丙酮酸2 g/L时发酵液黏度为16.0 Pa·s,此时胶体性能最好。     

Anthocyanin and colour evolution during maturation of four port wines: effect of pyruvic acid addition

The formation of vitisin A‐type compounds has been studied in four maturing fortified red port wines stored for 29 weeks at 15 °C. The anthocyanin concentrations were determined by high‐pressure liquid chromatography (HPLC), and colour changes were monitored by spectrometric measurements. The losses of anthocyanins followed first‐order reactions, and the concurrent formation of polymeric pigments was demonstrated. Vitisin A‐type compounds were found to be in low concentration in these four naturally maturing fortified wines. The addition of pyruvic acid to the wines led to the formation of large concentrations of vitisin A derivatives. Up to 23 mg litre⁻¹ vitisin A derivatives (vitisin A and its acylated forms acetylvitisin A and p‐coumarylvitisin A) could be determined. Owing to their greater colour expression and greater stability than malvidin 3‐glucoside, these new anthocyanins were shown to play an important role in the colour quality of the wines. An analytical survey of 32 port wines matured for between 2 and 6 years showed that vitisin A‐type compounds were the main, and sometimes the only, anthocyanins present. © 2000 Society of Chemical Industry