Vitisin A content in Chilean wines from Vitis vinifera Cv. Cabernet Sauvignon and contribution to the color of aged red wines

Vitisin A was prepared from malvidin 3-glucoside and pyruvic acid in model wine medium, isolated by countercurrent chromatography, and purified by preparative high-performance liquid chromatography (HPLC). The synthesized compound was used as a reference standard to quantify vitisin A in Chilean wines from Vitis vinifera cv. Cabernet Sauvignon, including a vertical row of wines from the same vineyard over 16 years. Maximum vitisin A content was reached within the first year of storage. Importantly, up to half of the initial amount of vitisin A in young wines was still present in 15 year old wines. Although vitisin A was found to be much more stable as compared to other monomeric C-4 underivatized anthocyanins, it also slowly degrades after reaching its peak concentration. The "color activity concept" was applied to vitisin A, malvidin 3-glucoside, malvidin 3-(6' '-acetylglucoside), and polymeric pigments isolated by countercurrent chromatography in order to estimate their contribution toward the overall color expression of wines. It was found that vitisin A is only a minor contributor to the visually perceived color of aged red wines (color contribution approximately 5%). The major contributor is the polymeric fraction (color contribution approximately 70-90%).     

In vitro interaction between ochratoxin A and different strains of Saccharomyces cerevisiae and Kloeckera apiculata

The interaction of ochratoxin A (OTA) and 20 yeast strains of Saccharomyces cerevisiae and Kloeckera apiculata during alcoholic fermentation was studied. Levels of OTA were determined in the fermentation liquid and in the yeast cells solid using a high-performance liquid chromatography system with a fluorescence detector. Yeast cells do not adsorb OTA, and for all yeasts, OTA levels did not affect the alcoholic fermentation. Some yeast strains reduced levels of OTA, whereas other strains did not show any effect demonstrating that OTA level reduction is not a genus species characteristic but a strain trait.     

Protein haze formation in white wines: effect of saccharomyces cerevisiae cell wall components prepared with different procedures

Cell wall material was extracted by five different methods from an oenological strain of Saccharomyces cerevisiae. Enzyme preparations containing beta-glucanase activity (Zymolyase, Glucanex, and Finizym 250L) allowed a better extraction yield compared to that of dithiothreitol (DTT) and EDTA. The yeast extracts were only soluble in part in wine. The wine-soluble fraction (WSF) of the five extracts, differing in both protein and sugar contents, when added in increasing amounts to white wine differently affected protein haze formation, as determined by the heat test, giving dose/response curves of different shapes. The curves obtained with the WSF derived from DDT and Zymolyase extracts showed a plateau value corresponding to 90% and 80% of wine haze reduction, respectively. In contrast, addition of the WSF derived from the other extracts resulted in increased turbidity with respect to the original wine. The mannoproteins (MP), isolated from each yeast extract by Concanavalin-A chromatography, gave dose/response curves showing shapes more similar among them than those obtained with the whole WSFs. The best wine stabilization was obtained with the MP of the DTT and Zymolyase extracts. The supernatants obtained after heating the MP of the different extracts were also tested. The stabilizing effect of the heat-stable MP (HSMP) was always larger than that of the corresponding total (un-heated) MP. The HSMP obtained starting from the DTT and Zymolyase extracts showed the best haze-protecting effect, which was, however, lower than that obtained with their corresponding WSF. This result suggests that wine protein stabilization by compounds of the yeast cell wall could be related, in addition to the action of the MP, also to the presence of other substances of different nature.     

Effect of different strains of Saccharomyces cerevisiae on production of volatiles in Napa Gamay wine and Petite Sirah wine

Napa Gamay grapes were fermented with four different strains of the yeast Saccharomyces cerevisiae (VL1, MI16, Fermirouge, and RA17). Petite Sirah grapes were fermented with seven different strains of the same yeast (BM45, Fermirouge, RA17, NI, CX3079, A350, and A796). Volatile compounds formed in the wines were analyzed by gas chromatography/mass spectrometry. Volatile compounds found in both wines were alcohols, esters, and acids, as well as some miscellaneous compounds. Isoamyl alcohol was the compound found in the highest relative amount with all four yeast strains in the Napa Gamay wines, followed by 2-phenyl ethanol, monoethyl succinate, and hexanoic acid. The relative amounts of isoamyl alcohol ranged from 30.84% (VL1) to 43.28% (RA17). Major volatile compounds found in Petite Sirah wines were isoamyl alcohol, 2-phenyl ethanol, 2-hydroxy ethyl propanoate, monoethyl succinate, and octanoic acid. The several esters, including 2-hydroxyethyl propanoate, may contribute to the fruity flavor of Petite Sirah wines. Overall, the S. cerevisiae yeast strains used to ferment Napa Gamay grapes and Petite Sirah grapes produced the same major components, with certain variations in formation levels.     

Aroma chemical composition of red wines from different price categories and its relationship to quality

The aroma chemical composition of three sets of Spanish red wines belonging to three different price categories was studied by using an array of gas chromatographic methods. Significant differences were found in the levels of 72 aroma compounds. Expensive wines are richest in wood-related compounds, ethyl phenols, cysteinil-derived mercaptans, volatile sulfur compounds, ethyl esters of branched acids, methional, and phenylacetaldehyde and are poorest in linear and branched fatty acids, fusel alcohols, terpenols, norisoprenoids, fusel alcohol acetates, and ethyl esters of the linear fatty acids; inexpensive wines show exactly the opposite profile, being richest in E-2-nonenal, E-2-hexenal, Z-3-hexenol, acetoin, and ethyl lactate. Satisfactory models relating quality to odorant composition could be built exclusively for expensive and medium-price wines but not for the lower-price sample set in which in-mouth attributes had to be included. The models for quality reveal a common structure, but they are characteristic of a given sample set.     

Heterocyclic acetals from glycerol and acetaldehyde in Port wines: evolution with aging

In Port wine, isomers of glycerol and acetaldehyde acetals have been found at total contents ranging from 9.4 to 175.3 mg/L. During oxidative aging, the concentrations of the 5-hydroxy-2-methyl-1,3-dioxane and 4-hydroxymethyl-2-methyl-1,3-dioxolane isomers increased with time showing a linear correlation (r > 0.95). The flavor threshold for the mixture of the four isomers was evaluated in wine at 100 mg/L. Thus, it is expected that they contribute to "old Port wine" aroma in wines older than 30 years. Experiments with model solutions and wine clearly demonstrated that SO(2) combines with acetaldehyde and blocks the acetalization reaction.     

Ferulic acid release and 4-vinylguaiacol formation during brewing and fermentation: indications for feruloyl esterase activity in Saccharomyces cerevisiae

The release of ferulic acid and the subsequent thermal or enzymatic decarboxylation to 4-vinylguaiacol are inherent to the beer production process. Phenolic, medicinal, or clove-like flavors originating from 4-vinylguaiacol frequently occur in beer made with wheat or wheat malt. To evaluate the release of ferulic acid and the transformation to 4-vinylguaiacol, beer was brewed with different proportions of barley malt, wheat, and wheat malt. Ferulic acid as well as 4-vinylguaiacol levels were determined by HPLC at several stages of the beer production process. During brewing, ferulic acid was released at the initial mashing phase, whereas moderate levels of 4-vinylguaiacol were formed by wort boiling. Higher levels of the phenolic flavor compound were produced during fermentations with brewery yeast strains of the Pof(+) phenotype. In beer made with barley malt, ferulic acid was mainly released during the brewing process. Conversely, 60-90% of ferulic acid in wheat or wheat malt beer was hydrolyzed during fermentation, causing higher 4-vinylguaiacol levels in these beers. As cereal enzymes are most likely inactivated during wort boiling, the additional release of ferulic acid during fermentation suggests the activity of feruloyl esterases produced by brewer's yeast.     

Pathway leading to the formation of anthocyanin-vinylphenol adducts and related pigments in red wines

On the basis of observations from Vitis vinifera cv. Pinotage wines and experiments performed in model wine medium, a new chemical pathway responsible for the formation of anthocyanin-vinylphenol adducts in red wines is described. Until now, these pigments have been considered to be reaction products of anthocyanins and vinylphenols, the latter being generated during fermentation by enzymatic decarboxylation of the respective cinnamic acids. The mechanism of the novel pathway, involving intact hydroxycinnamic acid and anthocyanin, is explained. Only cinnamic acids with electron-donating substituents on the aromatic ring, such as coumaric acid, ferulic acid, caffeic acid, and sinapic acid, undergo this conversion, as they stabilize an intermediately formed carbenium ion. Decarboxylation and oxidation of the pyran moieties are the final steps in the generation of the corresponding 4-vinylphenol, 4-vinylguaiacol, 4-vinylcatechol, and 4-vinylsyringol adducts of anthocyanins in red wine.     

Removal of ochratoxin A in red wines by means of adsorption treatments with commercial fining agents

The presence in wine of the fungal metabolite, ochratoxin A (OTA), represents a serious risk for consumer health. A variety of fining agents, including activated carbon, silica gel, potassium caseinate, egg albumin, and gelatin, was evaluated in relation to their abilities to remove OTA in fortified wines. Freundlich adsorption isotherms were used to model the adsorption behavior between ochratoxin A and the fining agent. Potassium caseinate and activated carbon were found to be the best fining agents that could be used to remove OTA in wine. Potassium caseinate removed up to 82% of OTA when used at 150 g/hL, whereas activated carbon showed the highest specific adsorption capacity due to a high surface area per mass and low adsorption of total polyphenols.     

温度对固定化酵母酒精分批发酵的影响及动力学模型

该文以甜高粱茎秆汁液为原料,探讨了温度（25～37 ℃）对甜高粱汁固定化酵母酒精分批发酵的影响,并对不同温度下固定化酵母乙醇发酵的动力学模型进行了研究。结果表明：温度的升高可以提高细胞生长速率,但过高的温度却阻碍了细胞的生长,从而影响了酒精的产量。应用Hinshelwood模型,分别对酒精发酵过程中细胞生长动力学和酒精合成动力学进行了模拟,得到25～34℃范围内不同温度下各种动力学参数。在此基础上,进一步研究了温度同细胞生长动力学参数之间的内在联系,得到酒精分批发酵过程中酵母细胞质量浓度的变化同温度以及底物质量浓度之间的一般关系式,验证试验结果表明,该模型具有很好的适用性。     

Identification of anthocyanin-flavanol pigments in red wines by NMR and mass spectrometry

Three newly formed Port wine pigments were isolated by Toyopearl HW-40(s) gel chromatography and semipreparative HPLC. Furthermore, the pigments were identified by mass spectrometry (LC/MS) and NMR techniques (1D and 2D). These anthocyanin-derived pigments showed UV-visible spectra different from those of the original grape anthocyanins. These pigments correspond to malvidin 3-glucoside linked through a vinyl bond to either (+)-catechin, (-)-epicatechin, or procyanidin dimer B3 [(+)-catechin-(+)-catechin]. NMR data of these pigments are reported for the first time.     

Improved organoleptic and nutritive properties of bakery products supplemented with amino acid overproducing Saccharomyces cerevisiae yeasts

Spontaneous yeast mutants isolated in continuous culture as resistant to toxic amino acid analogues, able to increase up to 40 times their free amino acid pool of Thr, up to 160 times their pool of Met, or up to 20 times their pool of Lys, were characterized with regard to properties of industrial interest. Growth rate, mu (h(-1)), and biomass yield, Y (g/L), of the amino acid overproducing mutants (AA(S)) were in many cases similar to those of the wild type, whereas their free amino acid content was substantially increased in laboratory and industrial media (molasses). Doughs fermented with 3% baker's yeast and 0.5% AA(S) mutants produced bakery products that displayed texture similar to those fermented with 3.5% baker's yeast, but the former had a considerable improvement of their taste and aroma. On the other hand, bread content of the essential amino acids Lys, Met, and Thr provided by yeast was also increased.     

Positive correlation between high levels of ochratoxin A and resveratrol-related compounds in red wines

The natural occurrence of ochratoxin A in red wines has been widely reported by several authors, as well as a that of group of stilbenes including cis- and trans-resveratrols and related glucosylated forms. In the present study 112 samples of retail red wines were collected from northern (17), central (46), and southern (49) Italy and were analyzed for both ochratoxin A and resveratrol-related stilbenes. The mean levels of total resveratrols and total piceids were 3.14 and 5.80 mg/L, respectively, whereas the ochratoxin A mean level was 0.64 microg/L. The Merlot wines showed the highest mean value of total stilbenes, followed by Negroamaro and Negroamaro blend, Aglianico, and Syrah, all with mean levels of >10 mg/L. Ochratoxin A was detected in 70, 59, and 100% of wine samples from northern, central, and southern Italy, with mean levels of 0.12, 0.07, and 1.36 microg/L, respectively. The highest values of ochratoxin A were recorded in Negroamaro- and Primitivo-based wine samples from southern Italy, showing also the highest content of stilbenes. In wine samples from southern Italy, a positive correlation was obtained between levels of ochratoxin A and total stilbenes (r = 0.74) as well as between ochratoxin A and total resveratrols (r = 0.50) and between ochratoxin A and total piceids (r = 0.74). These results suggest that toxic levels of ochratoxin A in red wine may be, at some extent, counterbalanced by the beneficial effects of resveratrol derivatives. Further investigation should be warranted in this regard.     

不同糖质原料和菌株固态发酵制取乙醇的特性比较

利用固态发酵技术直接将糖质原料转化为乙醇,过程简单且糖利用率高,已经成为近年来的一个研发热点。为寻求最适原料及菌种,该文对不同原料和菌株进行固态发酵产乙醇特性进行了比较研究:2种菌株为实验室自行筛选酵母菌株TSH和市场上常见的安琪酿酒酵母菌株AGL;3种糖料作物为甘蔗、甜菜和甜高粱。结果表明,无论对于TSH或者AGL,投入相同质量的原料,由于可发酵总糖初始含量的不同,最终乙醇产量最高为甜菜,甜高粱次之,甘蔗最低。为消除原料初始糖含量不同造成的影响,该文比较了消耗单位质量糖分所生成的乙醇质量,发现对于2种菌株甜高粱都具有最高的乙醇/糖转化率,说明甜高粱茎秆中糖分用于生成乙醇的比率较多,而用于维持酵母菌自身生长繁殖及副产物生成的糖分较少。比较2菌株的发酵能力,对于甜高粱TSH表现出较为明显的发酵优势:发酵周期比AGL缩短了近6h,且乙醇/糖转化率略高于AGL;而对于甘蔗和甜菜原料,TSH的乙醇产率与AGL相差不大;说明自行筛选的TSH菌种对多种糖料发酵底物具有广泛适用性,具有良好的工业应用前景。该文对利用糖质原料固态发酵生产燃料乙醇工艺的工业化实现及改进具有一定的借鉴意义。     

Geographic classification of spanish and Australian tempranillo red wines by visible and near-infrared spectroscopy combined with multivariate analysis

Visible (vis) and near-infrared (NIR) spectroscopy combined with multivariate analysis was used to classify the geographical origin of commercial Tempranillo wines from Australia and Spain. Wines (n = 63) were scanned in the vis and NIR regions (400-2500 nm) in a monochromator instrument in transmission. Principal component analysis (PCA), discriminant partial least-squares discriminant analysis (PLS-DA) and linear discriminant analysis (LDA) based on PCA scores were used to classify Tempranillo wines according to their geographical origin. Full cross-validation (leave-one-out) was used as validation method when PCA and LDA classification models were developed. PLS-DA models correctly classified 100% and 84.7% of the Australian and Spanish Tempranillo wine samples, respectively. LDA calibration models correctly classified 72% of the Australian wines and 85% of the Spanish wines. These results demonstrate the potential use of vis and NIR spectroscopy, combined with chemometrics as a rapid method to classify Tempranillo wines accordingly to their geographical origin.     

长期培肥下红壤有机碳组成与团聚体稳定性的关系

为研究培肥措施对红壤有机碳组成和团聚体稳定性的影响,以及有机碳与团聚体稳定性的关系,通过长期定位实验,选取不同施肥措施(未培肥CK、化肥NPK、化肥+秸秆NPKS和粪肥AM)下的典型红壤为研究对象,分析土壤有机碳组成和团聚体稳定性差异,揭示3种施肥措施下不同层次的有机碳组成和团聚体稳定性的变化规律.结果表明,3种施肥措施均可以提高土壤表层(0 ～25 cm)有机碳质量分数(尤其是颗粒有机碳),其中AM效果最显著,NPKS次之.不同施肥措施下红壤不同层次的团聚体稳定性顺序为AM＞ NPKS＞ NPK＞ CK.与CK相比,AM处理对表下层(5～15 cm)土壤的总有机碳和团聚体稳定性的提高效果最显著.回归分析表明,颗粒有机碳(POC)与湿筛法平均质量直径以及快速湿润(FW)、慢速湿润(SW)、预湿润震荡(WS)3种处理的平均质量直径的相关性最好(R2 =0.79、0.80、0.66、0.81),说明相对于其他组分,颗粒有机碳更有利于降低消散作用以及抵抗机械破碎进而增强团聚体稳定性,是间接评价土壤团聚体稳定性的良好指标.     

Evidence for an extracellular acid proteolytic activity secreted by living cells of Saccharomyces cerevisiae PlR1: impact on grape proteins

In this work, Saccharomyces cerevisiae PlR1, a strain isolated from Pinot noir grapes in the Champagne area, was shown to secrete an acid proteolytic activity against bovine serum albumin. This proteolytic activity was detectable in cell-free culture supernatants at the beginning of the exponential growth phase and increased with yeast growth. Using a zymography method, only one protease band with a molecular mass of 72 kDa was observed. This extracellular proteolytic activity was detected in the pH range from 2 to 4 with a maximal value at pH 2.5 and 38 °C and was completely inhibited by pepstatin A. The secretion of this protease did not need any protein inducer and seemed to be insensitive to nitrogen catabolic repression. S. cerevisiae PlR1 was also able to secrete this proteolytic activity during alcoholic fermentation, and it was found to be active against grape proteins, with a molecular mass around 25 kDa, at optimal conditions of 38 °C, pH 3.5.     

Determination of antioxidant power of red and white wines by a new electrochemical method and its correlation with polyphenolic content

A new method for measuring the antioxidant power of wine has been developed based on the accelerated electrochemical oxidation of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS). The calibration (R = 0.9922) and repeatability study (RSD = 7%) have provided good statistical parameters. The method is easy and quick to apply and gives reliable results, requiring only the monitoring of time and absorbance. It has been applied to various red and white wines of different origins. The results have been compared with those obtained by the total antioxidant status (TAS) method. Both methods reveal that the more antioxidant wines are those with higher polyphenolic content. From the HPLC study of the polyphenolic content of the same samples, it is confirmed that there is a positive correlation between the resveratrol content of a wine and its antioxidant power.     

Gene expression profiles of yeast Saccharomyces cerevisiae sod1 caused by patulin toxicity and evaluation of recovery potential of ascorbic acid

Patulin (PAT) is a fungal secondary metabolite and exhibits various toxicities including DNA damage and oxidative stress. These toxicities are eased by ascorbic acid (AsA). Although a number of studies regarding the mitigating effect of AsA against PAT toxicity have been reported, a comprehensive study about gene expressions is currently underway. Here, we carried out a detailed evaluation of PAT toxicity by co-incubation with AsA using the superoxide dismutase (SOD) mutant. DNA microarray results extracted the alterations in iron transporter and Fe/S cluster assembly genes; some of the genes that constitute the cellular iron transporter systems remained dysfunctional even in the presence of AsA. Meanwhile, AsA treatment reduced the alterations of G1/S phase cell cycle regulation genes. These results suggest that oxidative stress-derived DNA damage still exists, although AsA treatment effectively reduces PAT toxicity. This implies that a combined condition is required for complete blockade of PAT toxicity.     

Effect of EDTA alone and in combination with polygodial on the growth of Saccharomyces cerevisiae

The antifungal activity of ethylenediaminetetraacetic acid (EDTA) against Saccharomyces cerevisiae was significantly affected by various conditions such as inoculum size, pH, and metal ions (Mg(2+), Ca(2+)). EDTA was found to be effective against this yeast at the inoculum size of 10(5) colony forming units (CFU)/mL with the minimum inhibitory concentration of 400 mug/mL and the minimum fungicidal concentration of 6400 mug/mL, but it was not effective at 10(7) CFU/mL up to 6400 mug/mL. The fungicidal activity of EDTA against S. cerevisiae was significantly enhanced in combination with polygodial. Isobolograms, fractional inhibitory concentration, and fractional fungicidal concentration indices were used for evaluating the interaction between combined compounds. This synergistic effect is likely due to polygodial's destructive action on the cellular membrane, which facilitates the transmembrane transport of foreign compounds (EDTA) into yeast cells. Once inside the cells, EDTA forms chelation with divalent metals such as Mg(2+) and Ca(2+), which are required by various essential enzymes.