• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
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    • 专利摘要:
    Procedure for the treatment of musts and wines and product for its implementation. The procedure for the treatment of musts and wines of the invention consists of the addition of lysates of lactic acid bacteria in the refining or aging phase of the wine in order to reduce its astringency, an addition that also involves an increase in nitrogenous matter, at the same time that the negative Z potential of lactic acid bacteria and the ability to bond with the cations K+ and Ca2+ favors their retention, preventing their binding with tartaric acid as well as the formation of crystals of potassium bitartrate and neutral calcium tartrates, thus stabilizing the wine before the precipitate of tartrate salts, lysates that can also be used favoring the wine clarification process, being able to present the lactic bacteria lysate in liquid format or dry powder. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2889074A1
    申请号:ES202030524
    申请日:2020-06-04
    公开日:2022-01-11
    发明作者:Yoldi David García；Sáez Elisa Pérez；Goñi Ignacio López；Gómez Rafael Salgado
    申请人:Inbiolev SL；
    IPC主号:
    专利说明:

    [0002] Procedure for the treatment of musts and wines and product for putting it into practice
    [0004] TECHNICAL SECTOR
    [0006] The present invention refers to a procedure intended to be applied as sub-processes in the wine-making process itself.
    [0008] More specifically, the process consists of the use of lactic acid bacteria lysates for oenological applications such as the reduction of astringency in wines or nutrition in fermentation processes in musts and wines. In the same way and due to the nature of negative charges in the cell wall of lactic acid bacteria, its application would be extended to the tartaric stabilization of wines.
    [0010] Finally, the planned process is also applied to the clarification of wine.
    [0012] The object of the invention is therefore to provide a process that allows the production of wines with high quality standards that satisfy the demands of an increasingly demanding consumer.
    [0014] The invention also relates to the product for carrying out the process described.
    [0016] In short, the invention is applicable in the food sector, especially in the wine industry, without ruling out other industries such as beer, cider or any other beverage where it is intended to reduce astringency, stability or non-formation of salts present in other fermentative matrices, as well as in the clarification or reduction of turbidity in beverages.
    [0018] BACKGROUND OF THE INVENTION
    [0020] With regard to the problem of astringency in wines, today, due to climate change and the increasing number of hours of annual sunshine, grapes experience a natural dehydration that causes a rapid increase in sugars that leads to an increase in potential alcoholic strength without the grapes having acquired phenolic maturity. This is a problem that leads to greenness and astringency in the fermented musts from these grapes.
    [0022] Viticulturists and wineries are sometimes forced to pick the grapes early so that the alcoholic strength is not too high, without giving time for the polyphenols in the grape seeds and skins to mature and lose their greenness and astringency. .
    [0024] To alleviate this problem in the oenological world, products are used whose application consists of masking the astringency and greenness caused by the polyphenols that make up the grape that have not reached their optimum ripening or polymerization over time.
    [0026] Today the products used are based on mannoproteins from the Saccharomyces genus and cerevisiae species. The action of these mannoproteins consists of polymerizing or binding polyphenols to each other so that they do not have as many reactive groups in the wine so that they stick or unite with the saliva proteins, condense and give sensations of astringency or dry mouth.
    [0028] Regarding nutrition in the production process in the wine industry, lactic acid bacteria lysates, made up of amino acids, proteins, polysaccharides and specific nucleic acids, represent a unique source of nitrogenous compounds for fermentation processes in the food industry, in particular, in the wine industry. Thus, the amino acids are aromatic precursors and the proteins and polysaccharides intervene in the sensory matrix, providing structure and weight in the final product as well as increasing the glyceric and clean sensations by reducing the sulfur notes.
    [0030] For its part, and with regard to tartaric stabilization, tartaric acid is one of the main acids in wines, which can be partially insolubilized by the presence of calcium and potassium cations, forming salts. The solubility of these salts decreases during alcoholic fermentation, as well as by cooling the wine. During the conservation of the wines and especially during the winter a spontaneous insolubilization of tartrates takes place, but in spite of this the wines can contain in A sufficient quantity of these salts dissolves and new precipitations can occur when the wine is bottled, which can be avoided with a timely stabilization treatment applied before bottling. In addition, due to the excess of potassium that is frequently found in vineyards around the world, they are more unstable wines, appearing precipitation of potassium bitartrate (THK). The presence of these small potassium bitartrate crystals at the bottom of the bottle generally appears as a consequence of a decrease in temperature, but they do not constitute a defect. This precipitation is found in less quantity or is not found in wines that have gone through a stabilization process.
    [0032] Although the presence of these crystals does not negatively affect the taste of the wine, for consumers the presence of these crystalline precipitates denotes a defect and they would reject the wine since they do not know the origin of the precipitate, generating distrust in the brand.
    [0034] Among the most widespread treatments at present, we can mention systems that insolubilize and eliminate tartrates from wine (cold and reverse osmosis), systems that prevent tartaric precipitation (CMC, mannoproteins and metatartaric acid).
    [0036] Stabilization techniques are not well developed, since in addition to eliminating the problem of tartaric precipitation, favorable compounds for wine such as aromas and polyphenols can be eliminated, thus modifying the organoleptic characteristics of the wine. For this reason, it is necessary to delve into these techniques to improve the quality of the wine after a tartaric stabilization process.
    [0038] Tartaric acid (TH2) is characteristic of grapes. Its content is variable and tends to be lower in southern regions where bunch exposure temperatures are higher. Generally, it is TH2 that gives the wine its characteristic pH value (3.00-3.50), which gives it a pleasant sensation of freshness, while a possible deficiency makes the wine flat and without body. As is known, the TH2 content tends to decrease spontaneously over time due to the precipitation of its calcium (Ca) and K salts. From this follows the importance of a tartaric stabilization treatment to avoid these precipitations in the bottle. .
    [0039] As mentioned, during the fermentation phase THK is formed, and in addition, calcium tartrate (CaT) can also be formed. The natural tartaric acid in wine (TH2) is capable of combining with the (K) present in grapes. The corresponding salt (THK) is soluble in grape must, but much less when alcohol is present, after fermentation. Naturally, THK sediments. If no action is taken while the wine is still in the tank, the THK crystals will settle in the bottles.
    [0041] Therefore, metals, sulfates, proteins (for example of a mannoprotein nature) and polyphenols can form complexes with free tartaric acid and potassium ions, inhibiting the formation of THK.
    [0043] The complexes formed are mainly between polyphenols and proteins with tartaric acid.
    [0045] The binding of free tartaric acid prevents the formation of bitartrate.
    [0047] Mannoproteins or glycoproteins are adsorbed on the crystal surface of the crystal nucleus inhibiting its growth.
    [0049] Among the current tartaric stability treatments, one of the best known is cold treatment, although what is currently being sought are alternative, cheaper treatments with better results.
    [0051] Cold treatment requires a high investment in isothermal tanks, cold equipment, high electrical consumption, long duration of treatment as well as requiring subsequent filtration.
    [0053] Reverse osmosis is a technique that involves putting a wine in contact with a reverse osmosis module, which has a membrane with a suitable molecular cut-off (smaller than that of ultrafiltration). It is achieved that through this, in the fraction known as permeate, only water and solutes with lower molecular weight (for example, acetic acid) pass through, being retained in the original matrix (fraction called retained) the wine components most characteristic, which are usually those with the highest molecular weight.
    [0054] The wines are passed through reverse osmosis equipment, where part of the water they contain is temporarily removed, resulting in concentrated wines and producing a significant insolubilization of the tartrates that can be activated with a complementary cold treatment. Once separated, the initially separated water is restored, thus resulting in stabilisation.
    [0056] The addition of metatartaric acid is another method that is used on finished wine and involves a cost in product and handling as well as no guarantee of stabilization over long periods. The maximum legal dose of 10g/hL.
    [0058] Metatartaric acid is obtained by controlled vacuum dehydration of D-tartaric acid at heat between 150 and 170°C. It is a polyester of tartaric acid whose main components are the monoester and the ditartric diester.
    [0060] Its effectiveness depends on its degree of esterification. In aqueous solutions such as wines, the protective effect is not stable, since over time this product tends to hydrolyze slowly, forming tartaric acid again, which can include even greater precipitation. For this reason, this product limits its use for fast-rotation wines and in any case not intended for storage and conservation, since it prevents tartaric precipitation for a maximum of 9 months. This loss of efficiency is greater the higher the temperature and the lower the pH. It is used at the end of fermentation and just before bottling. It should not be used in hot bottling, because its hydrolysis is enhanced at high temperatures. Metatartaric acid prevents the growth and precipitation of crystalline nuclei, thus inhibiting the appearance of tartaric precipitates in the bottle. The mechanism of inhibition of metatartaric acid on tartrates in wine is explained by the formation of a barrier around the crystallization nuclei, which prevents the approximation of insolubilized tartrate molecules and therefore the formation or rather the growth of crystals. . When the protection is insufficient and the tartrate insolubilization conditions are adequate, then a partial inhibition phenomenon can occur, thus forming anomalous tartrate crystals with irregular shapes.
    [0061] Another methodology is carboxymethylcellulose (CMC), which has been questioned for several years until it was authorized for oenological use (Resolution OENO 2/2008). It involves a somewhat complicated preparation methodology.
    [0062] CMC was authorized by the European Community in 2009 as an alternative to traditional electrodialysis or cold treatments used for tartaric stabilization of wines. It can completely replace cold treatment. Prevents the formation of THK crystals, with a dose of 5 g/hL. It involves cost in product and handling.
    [0064] This cellulose derivative, extracted only from wood intended for oenology (to avoid the risk of product contamination from a source of genetically modified organisms (GMOs) that may come from cotton) and from sustainably managed forests, is It has been used for decades in a large number of food preparations (E468). It has the advantage of being completely neutral at an organoleptic level and very effective in stabilizing wines against THK. Its action is known to be effective for at least 4 years. Although it is an expensive technique, several researchers have shown that CMC is an excellent alternative; much superior to metatartaric acid that hydrolyzes over time.
    [0066] CMC is a crystallization inhibitor. Its mechanism of action is still the subject of hypotheses. It is assumed that, as soon as the crystals begin to form, the CMC is deposited on some faces and the K or bitartrate ions can no longer make the crystal grow. It is added after filtration.
    [0068] It is recommended to use CMC especially in white and rosé wines. In red wines, there is a risk of a loss of colloidal materials, which is why it is necessary to carry out specific tests to avoid problems after bottling. It can be said that the richer the wines are in polyphenols, the greater the risk of haze formation.
    [0070] Like any other additive technique, CMC is active on the tartaric acid salt up to a certain limit. Compared to THK, with CaT it is less active, but there is activity. It is necessary to know that if the wine is unstable with respect to the two tartaric salts THK and CaT and that if the instability with respect to THK is very high, the CMC added to the wine, it will be mobilized to prevent the formation of these crystals and therefore it will be less available to inhibit the crystallization of CaT.
    [0072] Due to its molecular structure, CMC behaves in a similar way to metatartaric acid, as a protective colloid; binds to the surface of dissolved THK preventing crystal growth; but its advantage is that it is not sensitive to temperature.
    [0074] The use of mannoproteins is another methodology, so that yeast mannoproteins constitute the second largest family of polysaccharides naturally present in wine.
    [0076] In April 2006, the EC authorized the use of mannoproteins in the wine sector (OIV, 2006). It represents a natural alternative to current treatments. Its use allows to reduce the consumption of water and energy. The presentation in liquid form allows its direct addition to the wine before bottling, thus offering optimum treatment efficiency and flexibility.
    [0078] The preventive action of mannoproteins on the tartaric crystallization of wines is attracting more and more interest. However, little is known about the diversity of yeast mannoproteins and their effect on wines. These differences are the origin of its functional properties and its effects on the wine, which range from improving volume in the mouth to inducing tartaric stability.
    [0080] Mannoproteins inhibit crystal nucleation (initial stage of crystal formation), their effect on crystal growth is less important. Therefore, the protection of a wine against tartaric instability is effective only in the absence of crystals.
    [0082] This use of mannoproteins proves to be very effective against tartaric crystallization in wines for a period of at least 18 months.
    [0084] Finally, and with regard to the fining process, fining agents are traditionally applied during wine production to obtain a smoother product. bright and clarified as a result of the elimination of the particles responsible for turbidity. These agents are also beneficial in softening tannic intensity, improving mouthfeel perception, and improving filterability and stabilization of the wine.
    [0086] Clarification also reduces protein instability, which is a great improvement, as the presence of unstable proteins in wines is currently a major concern for winemakers. This process is expected to remove suspended particles and certain compounds responsible for oxidation reactions and the undesirable astringency of wine, enhancing its organoleptic characteristics and improving its visual appearance. In this case, the color, brightness and clarity of the wine are crucial sensory attributes that are highly valued by consumers and can make a wine accepted or rejected. Depending on the type of wine and the desired effect, it is necessary to choose the right fining agent to be applied.
    [0088] The most widely used clarifying agents in the wine industry can be obtained from animal proteins, vegetable proteins and inorganic compounds. These proteins reveal a great diversity of molecular masses, isoelectric points and/or surface charge densities, and depending on the conditions of each wine, they will interact differently with specific components of the wine.
    [0090] Protein clarifying agents have the ability to form complexes with wine tannins, creating negatively charged hydrophobic colloids. In the presence of metallic cations, these colloids become insoluble and tend to precipitate, in a process of flocculation. On the contrary, proteins that do not interact with tannins have a tendency to combine with particles in suspension or in colloidal solution, which are mostly negatively charged, which also leads to their sedimentation. Other proteins such as caseins are insoluble in the low pH of wine, so they tend to coagulate and flocculate. However, to induce its precipitation and wine clarification, the presence of tannins is required. Simultaneously, they are able to bind and therefore eliminate the phenolic compounds that affect the color and flavor of wines.
    [0092] Although these clarifying agents appear to be highly effective in stabilizing and clarifying wine, some of them may endanger the health of the consumer.
    [0093] For example, casein from milk and ovalbumin from chicken egg white, both of origin. This type of clarifying agents belongs to the "hidden allergens", which are allergenic ingredients present in complex foods and that cannot be recognized by the common consumer. This fact raises various concerns in the consumer, since there is a risk of food intolerances and allergic reactions, which leads consumers to be more careful with the food they consume. Since these issues can endanger the health of the consumer, the European Union applied a regulation (EU Regulation No. 1169/2011) that requires the labeling of all substances and products that can cause these allergies and intolerances. Subsequently, Regulation (EC) No.
    [0094] 579/2012 to make it mandatory to label all milk-based, milk-based or egg-based fining treatments, and egg-based products used in grape must and wine. The application of these regulations is extremely important, since they provide all the information to the consumer, allowing a conscious choice of the consumer.
    [0096] On the other hand, non-protein fining agents are also currently used in wines, such as polyvinylpolypyrrolidone (PVPP), bentonite, activated carbon and chitosan. PVPP acts in a similar way to protein fining agents and exhibits adsorption very selective of phenolic substances, specifically anthocyanins and catechins, reducing their quantity in the wine. For example, polyphenols are essential components of red wines; however, when excessive, they can cause undesirable characteristics such as bitterness and astringency. Although it is not usually applied to red wines, PVPP improves the color and brightness of the wine, also reducing its bitterness, without removing the wine's aroma. As for bentonite, it is a cation exchanger clay, which carries a net negative charge and therefore interacts electrostatically with positively charged proteins at wine pH. When added to wine, it removes unstable proteins, however, the action of bentonite appears to be non-protein specific as it also removes other species or charged aggregates that may be beneficial molecules for wine aroma and color. contributes to the loss of sensory quality that is often claimed in bentonite treated wines, furthermore, treatments with this fining agent can mean a large loss of wine volume, 5% to 20%, as bentonite lees. In addition, bentonite poses a series of environmental problems, since it is associated with occupational health risks, including dust inhalation, and disposal of hazardous bentonite waste.
    [0098] Due to the health problems related to animal protein agents and the high environmental impacts of bentonite and PVPP, the need to find a different approach based on substances that do not endanger the health of the consumer has become a great concern. challenge. To overcome this concern, yeast protein extracts (YPE), obtained from endogenous wine and grape yeasts, appear to be a potential alternative. In fact, several studies show that the application of new clarification treatments based on yeast derivatives improve wine quality by reducing its turbidity and astringency.
    [0100] Yeast protein extracts are not classified as food allergens by European law and, since 2013, the clarification of grape must and wine is allowed within the European Union (EC Regulation No. 144/2013). The application of YPEs as fining agents is also permitted by the International Organization of Vine and Wine (OIV) [resolutions OIV-OENO 416-2011 and OENO 417-2011] and established a monograph [resolution OIV-OENO 452-2012 ] that offers a new alternative to the wine sector.
    [0102] Yeast protein extracts can be obtained from different components of yeasts, such as the cytoplasm, vacuole or cell wall, and are applied during the winemaking process for various purposes. For example, in several species of yeast, including Saccharomyces cerevisiae, protein extraction occurs from the cell wall, because it is composed of an outer layer of highly glycosylated mannoproteins covalently bound to an amorphous matrix of p-glucans in the outer layer. of the wall. However, mannoproteins show a lower binding affinity to wine tannins compared to non-glycosylated proteins. Therefore, because the presence of polysaccharides and mannoproteins can inhibit the precipitation of fining agent proteins with wine compounds, particularly tannins.
    [0104] Therefore, the development of protein extracts derived specifically from the soluble extract after total degradation of the cytoplasmic content may be crucial. and removal of cell walls.
    [0106] EXPLANATION OF THE INVENTION
    [0108] The procedure that is recommended for the reduction of astringency, nutrition in wine fermentation processes, tartaric stabilization and wine clarification fully satisfactorily solves the previously exposed problems, in each and every one of the previously mentioned aspects.
    [0110] The use of lactic acid bacteria lysates is natural since it is a constituent part of the grape itself, but never before have lactic acid bacteria lysates been used to overcome astringency, nutrition and clarification problems in the food sector.
    [0112] To this end, and more specifically, the method of the invention consists of the application and use of lactic acid bacteria lysates in the food sector and in particular in the oenological sector.
    [0114] This invention is based on the fact that lactic acid bacteria, naturally present in wine, have a low Z potential (negative charge) in their membrane, presenting a high reactivity, with an affinity to bind positively charged free tannins in such a way that they are retained. preventing its link with saliva proteins, also negatively charged.
    [0116] It has been found that lactic acid bacteria lysates have a higher affinity for tannin binding. More specifically, compared to yeast lysates, lactic acid bacteria lysates have a higher negative Z potential or a higher negative charge expressed in millivolts, since, unlike yeast, they have two envelopes: the peptidoglycan and the cell wall. , both negatively charged, unlike yeasts that only have the cell wall.
    [0118] Thus, the use of lactic acid bacteria lysates, which to date had never been postulated for these applications or had never been valued, has a competitive advantage over those currently used, such as yeast lysates or autolysates or their fractions. cell wall mannoproteins.
    [0119] Consequently, the invention focuses on the use of lactic acid bacteria lysates, lactic acid bacteria autolysates or peptidoglycan proteins (which the yeast does not possess) or lactic acid bacteria cell wall for use and use in applications such as astringency, tartaric stabilization or clarification.
    [0121] Experimentally, to solve the problem of astringency, it has been possible to verify how this process significantly reduces astringency both at the quantitative level of free tannins (direct measurement) and at the qualitative level, organoleptically, comparing it with a witness or negative control and also with products currently used commercial or positive controls.
    [0123] The use of lactic acid bacteria lysates to correct astringency resolves in an efficient and timely manner what has been raised so far by the wine industry.
    [0125] The product described as lactic acid bacteria lysate will be presented either in liquid or dry powder format.
    [0127] On the other hand, the use of lactic acid bacteria lysates solves, just like yeast lysates, the problem of nutrition in wines, having verified how the use of lactic acid bacteria lysates offers a singular nitrogenous matter never before available for this purpose, which offers regular and safe fermentative kinetics, enhancing at the same time the sensory profile, producing limpidity of the aromatic matrix.
    [0129] Regarding nutrition in the production process in the wine industry, lactic acid bacteria lysates, made up of amino acids, proteins, polysaccharides and specific nucleic acids, represent a unique source of nitrogenous compounds for fermentation processes in the food industry, in particular, in the wine industry. Thus, the amino acids are aromatic precursors and the proteins and polysaccharides intervene in the sensory matrix, providing structure and weight in the final product as well as increasing the glyceric and clean sensations by reducing the sulfur notes.
    [0131] Due to their high negative charge of the Z potential, lactic acid bacteria lysates are capable of tartar stabilizing wines, due to their ability to bind with K+ and Ca2+ cations favor this union, in such a way that the potassium cation and the calcium cation are retained, preventing their bond with Tartaric Acid as well as the formation of crystals of potassium bitartrate and neutral calcium tartrates, thus stabilizing the wine before the precipitate of tartrate salts.
    [0133] Finally, and with regard to the clarification process, and as has been commented on in the previous sub-processes, the high negative charge that lactic acid bacteria lysates have, allows for the purification of certain proteins or the obtaining of of protein complexes once the cell rupture of the lactic bacteria has been carried out and the obtaining of protein complexes obtained from the clarified part of the lyses generated with different molecular screening methods for its application in the clarification of wines due to union and successive sedimentation through interaction with positively charged wine proteins at that wine pH that can form larger structures of high molecular weight and that can precipitate together with the non-binding of some cations present in the wine.
    [0135] These different lysates, proteins, glycoproteins of lactic acid bacteria, as well as mannoproteins in yeasts, can act as an effective substitute/alternative for clarifying agents, stabilizers at the protein level once the wine is finished, due, as mentioned above, to the fact that the bacteria lactic bacteria have a greater negative Z potential and therefore a greater net negative charge, so the use of either total lactic acid bacteria lysates or singular protein or glycoprotein parts that can be extracted from them for clarification treatments it will be extremely beneficial.
    [0137] The product described as lactic acid bacteria lysate will be presented either in liquid or dry powder format.
    [0139] In short, the novelty of the invention focuses on the use of many different genera, species and strains of lactic acid bacteria present in wine, specifically the use of proteins, glycoproteins, polysaccharides or complete cell wall lysates, peptidoglycan and cytoplasmic content. in lactic bacteria for its use against astringency, clarification, tartaric stabilization already mentioned and that is fundamental thanks to the high power of net negative charge of lactic bacteria against yeasts.
    [0140] In addition, the fact of valuing the use of lactic bacteria is their consideration of food grade, allowed in wine and naturally present in wine or grapes, the intrinsic composition of the product is not artificially modified.
    [0142] DESCRIPTION OF THE DRAWINGS
    [0144] To complement the description that will be made below and in order to help a better understanding of the characteristics of the invention, according to a preferred example of its practical embodiment, an example of reducing astringency by means of of lactic acid bacteria lysates and another of nutrition in a fermentation process, accompanied as an integral part of said description, a set of plans where, by way of illustration and not limitation, the following has been represented:
    [0146] Figure 1. Tannic acid concentration (g/L) after wine treatment for 28 days. Results shown are the mean of two replicates performed under the same conditions ± standard deviation (error bars). Asterisks (*) indicate statistically significant differences ap<0.05 (*), p<0.01 (**) or p<0.001 (***) between wine treated with mannoprotein (positive control) and lactic acid bacteria lysates compared to wine without treatment according to the Tukey test, it being possible to see how the lactic acid bacteria lysates decrease astringency to a greater extent due to their ability to reduce the amount of tannins.
    [0148] Figure 2. Fermentative kinetics of an oenological process with lactic acid bacteria lysate compared to current commercial proposals based on yeast fragments and compared to a control (C-), being able to verify how the fermentative kinetics are not modified when yeast-based nutrients are introduced. lactic bacteria lysates in relation to that obtained when commercial nutrients based on autolysates or yeast extracts are introduced.
    [0150] Figure 3. Tasting diagram of a must fermented with organic nutrition based on lactic acid bacteria lysates and organic nutrition commercial proposals based on yeast fragments, where it can be verified that the lactic acid bacteria lysates achieve equivalent or better profiles than those obtained with nutrients commercial employees.
    [0152] PREFERRED EMBODIMENT OF THE INVENTION
    [0154] In view of the figures outlined, and by way of example or study, the neutralizing effect of bacterial lysates against the reactive tannins of wine responsible for astringency is analyzed.
    [0156] In this sense, yeast lysates or lees have a neutralizing effect on tannins in the refining or aging phase on lees due to their rich composition of mannoproteins and other membrane and intracellular proteins. Bacterial cells differ in composition from yeast cells, therefore the aim was to determine if the proteins and peptidoglycans of bacteria are equally or more reactive against free radicals from tannins, causing their precipitation.
    [0158] To do this, the total protein concentration of the negatively charged lysates was determined and a finished wine was treated with the different bacterial lysates at equal protein concentration for one to four weeks. After the treatment time, the concentration of reactive tannin present in the treated wine compared to the untreated wine was determined.
    [0160] The refining treatment was carried out with finished red wine of the Tinta Fina variety.
    [0162] Due to the high concentration of reactive tannin naturally present in the seeds, one liter of wine was macerated for 3 days at 20°C with 40 crushed seeds and diluted with 4 liters of wine. The initial biochemical parameters of the wine-sample (matrix red wine) are indicated in Table 1.
    [0164] Table 1 Biochemical parameters of the parent red wine.
    [0166]
    [0167] Bacterial lysates were produced by heat treatment. The selected lactic bacteria are shown in table 2.
    [0169] Table 2 Species, strains and lysates generated for the study.
    [0174] The protein concentrations of each lysate were measured by the BCA method and the treatment dose of each lysate was adjusted according to the total protein concentration of a commercial lysate at a dose of 0.1 ml per liter of wine, whose total protein concentration is 23.7 milligrams of protein per milliliter of lysate. That is, the protein concentration with which the wine was treated is the same for all treatments with lysates.
    [0176] The treatment with the lysates was compared with respect to commercial controls of mannoprotein from yeast, at the same dose in total protein and was also compared with a control without treatment.
    [0178] The wine treatment was carried out in test tubes with 15 ml of wine in which the treatment was added and closed hermetically and without head space. The treatment was carried out in a climatic control chamber with a controlled temperature of 20 °C. Both the initial measurements of reactive tannin and the measurements after seven and twenty-eight days of treatment were carried out using the modified Llaudy method.
    [0180] The results shown have been obtained by comparing each treated sample with respect to the untreated sample. Analysis of variance (ANOVA) was used to detect statistical differences between samples using the GraphPad PRISM tool. The statistical significance of each one of them was considered at p<0.05 according to Tukey's test.
    [0182] The results obtained are the following:
    [0184] The wine sample processed with seeds to increase reactive tannin and the sensation of astringency had a tannic acid concentration of 0.56 g/L at initial time. At seven days, the untreated sample had a concentration of 0.54 g/L and at 28 days 0.48 g/L of tannic acid, that is, the tannins in the samples partially polymerized without the need for treatment.
    [0186] Table 3 Concentration and reduction of tannic acid in 7 days compared to the control corresponding to the wine without treatment (N.S; non-significant differences).
    [0190] Table 4 Concentration and reduction of tannic acid in 28 days with respect to the control corresponding to the wine without treatment (N.S; non-significant differences).
    [0191]
    [0192] As can be seen, due to the greater negative Z potential, the reduction in all cases with any genus of lactic acid bacteria resulted in a greater reduction in tannic acid compared to commercial high-end yeast mannoprotein lysate that is sold and recommended for this purpose.
    [0194] Therefore, expanding this fact, it can be concluded that the specific lysates used in the practical embodiment are in no way limiting, the invention being applicable to any lysate of lactic acid bacteria, autolysates of the same or purified protein fractions of peptidoglycan or lactic acid bacteria cell wall.
    [0196] Regarding the effect of lactic acid bacteria lysates on the reduction of tartaric crystallization, the "pear tube test" (M. Ruiz Hernández) was carried out as follows:
    [0198] - A volume of 100 cc of wine was taken in a tube.
    [0199] - It was frozen for 24 hours at -18 °C.
    [0200] - It was allowed to thaw and the sediment was measured.
    [0202] The wine is considered stable if the sediment concentration is less than 0.30 cc., having obtained in the treatment with lactic bacteria lysate a volume of precipitate of 0.20 cc., less than that obtained with the treatments of existing commercial mannoproteins for said application and notably lower than the precipitate obtained when no treatment was carried out.
    [0204] Finally, and with regard to the clarification process, experimentally it was also possible to verify how the use of this type of lysates allows the purification of certain proteins or the obtaining of protein complexes once the cell rupture of the lactic acid bacteria and the Obtaining protein complexes obtained from the clarified part of the lyses generated with different molecular screening methods for application in wine clarification due to binding and subsequent sedimentation through interaction with positively charged wine proteins at that pH of the wine that can form larger structures of high molecular weight and that can precipitate together with the union or not of some cations present in the wine.
    [0205] Consequently, these different lysates, proteins, glycoproteins of lactic acid bacteria, as well as mannoproteins in yeasts, act as an effective substitute/alternative for clarifying agents, stabilizers at the protein level once the wine is finished, due to the fact that lactic acid bacteria have a greater potential Z negative.
    权利要求:
    Claims (6)
    [1]
    1a.- Procedure for the treatment of musts and wines, characterized in that it consists of applying lactic acid bacteria lysates, lactic acid bacteria autolysates or purified protein fractions of peptidoglycan or lactic acid bacteria cell wall during the wine production process as a regulating agent of astringency, as an agent for nutrition in fermentative processes, as a tartaric stabilizing agent and/or as a clarifying agent.
    [2]
    2nd.- Procedure for the treatment of musts and wines, according to claim 1, characterized in that the species of lactic acid bacteria lysates used during the procedure are any or a combination of the following:
    • Leuconostoc mesenteroides.
    • Lactobacillus cacaonum.
    • Lactobacillus plantarum.
    • Lactobacillus brevis.
    • Lactobacillus psittaci.
    • Lactobacillus senmaizukei.
    • Lactobacillus hilgardii.
    • Lactobacillus plantarum.
    • Lactobacillus brevis.
    • Leuconostoc mesenteroides.
    • Lactobacillus cacaonum.
    • Pediococcus damnosus.
    • Lactobacillus lactis.
    • Oenococcus oeni.
    • Lactobacillus brevis.
    • Pediococcus cellicola.
    • Leuconostoc pseudoficulneum.
    [3]
    3 a.- Product for the treatment of musts and wines, according to the procedure of claim 1, characterized in that it consists of a product in liquid format or in dry powder, based on lactic acid bacteria lysates, lactic acid bacteria autolysates or purified protein fractions of peptidoglycan or lactic acid bacteria cell wall, intended to reduce astringency in wines.
    [4]
    4a.- Product for the treatment of musts and wines, according to the procedure of claim 1a, characterized in that it consists of a product in liquid format or in dry powder, based on lactic acid bacteria lysates, lactic acid bacteria autolysates or purified protein fractions of peptidoglycan or cell wall of lactic acid bacteria, intended for nutrition in fermentation processes in musts and wines.
    [5]
    5.- Product for the treatment of musts and wines, according to the procedure of claim 1, characterized in that it consists of a product in liquid format or in dry powder, based on lactic acid bacteria lysates, bacteria autolysates lactic or purified protein fractions of peptidoglycan or cell wall of lactic acid bacteria, intended to stabilize wines tartaric.
    [6]
    6a.- Product for the treatment of musts and wines, according to the procedure of claim 1, characterized in that it consists of a product in liquid format or in dry powder, based on lactic acid bacteria lysates, lactic acid bacteria autolysates or purified protein fractions of peptidoglycan or cell wall of lactic acid bacteria, intended for wine clarification.
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