JP2022082114A - Method for producing wine - Google Patents

Abstract

To provide a method for producing a wine having enhanced redness, by performing alcohol fermentation with NYR20 strain, in the presence of sulfurous acid.SOLUTION: The present invention discloses a method for producing a wine having enhanced redness by using NYR20 strain (accession number: NITE P-03294), being the yeast to produce a red color. The method includes the step of performing fermentation by adding the yeast to a base liquid containing grape juice and sulfurous acid.SELECTED DRAWING: Figure 1

Description

There is an application for application of Article 30, Paragraph 2 of the Patent Law. It was released at the ASEV JAPAN 2019 Conference of the Japan Grape and Wine Society held from November 29, 1st to 30th of the same year. It was published in "Volume 30 Number 2 of the Journal of the Japan Grape and Wine Society", which was published on November 29, 1st year of Reiwa. It was released on the following website on August 31, 2nd year of Reiwa. https: // www. pref. Hiroshima. lg. jp / site / innovation / monodukurisaitaku. html https: // www. pref. Hiroshima. lg. jp / upgraded / attendment / 403131. pdf Reiwa Reiwa was released on the following website on October 22, 2nd year. http: // rdbv. fukuyama-u. ac. jp / view / 4D5TK /

The present invention relates to a method for producing wine.

The following Patent Document 1 describes a method for producing sake using the NYR20 strain. The NYR20 strain was obtained by treating yeast isolated from the flowers of Prunus verecunda with a mutagen. Patent Document 1 describes that red-colored sake is produced by brewing using the NYR20 strain.

Japanese Unexamined Patent Publication No. 2014-212782

In the method of Patent Document 1, rice jiuqu, water, rice, and a mash containing NYR20 strain are mixed to make mash, and the mash is fermented to produce sake. Sulfurous acid is not used in the production of sake.

In the wine manufacturing process, raw materials containing wine juice are used for the purpose of suppressing the odor of acetaldehyde generated during fermentation, preventing changes in flavor due to oxidation of the produced wine, and preventing the growth of germs during the fermentation process. Sulfurous acid is added to the liquid in the form of sulfurous acid salt, sulfurous acid gas, or sulfurous acid water.

It is unclear how much sulfite resistance the NYR20 strain has, and when the stress caused by sulfite and the stress caused by alcohol generated in the fermentation process are added in combination, a red pigment is produced and alcohol is also produced. It was unclear if it could be generated.

An object of the present invention is to provide a method for producing wine with enhanced redness by performing alcoholic fermentation using NYR20 strain (accession number: NITE P-03294) in the presence of sulfurous acid.

In the present invention, the NYR20 strain (accession number: NITE P-03294) is capable of performing alcoholic fermentation even in the presence of sulfite and ethanol, and has the ability to secrete a red pigment outside the cells. It is based on the discovery that it does not lose. The present invention is a method for producing wine with enhanced redness using a yeast NYR20 strain (accession number: NITE P-03294) that produces a red pigment, and is described above with respect to a raw material solution containing grape juice and sulfite. It is a method for producing wine including a step of adding yeast and performing fermentation, and the present invention solves the above-mentioned problems.

In the above-mentioned method for producing wine, in the step of fermenting, the yeast is added to the raw material solution containing grape juice so that the number of bacteria exceeds 1.6 × ¹⁰⁶ cells / mL, and fermentation is performed. It is preferably a process.

In the above method for producing wine, the grapes are white grapes, and after obtaining fruit juice from the white grapes, fermentation with the yeast is carried out using a raw material solution obtained by removing the peel of the white grapes from the fruit juice of the white grapes immediately. May be. Further, the grapes are white grapes, and the yeast is used as a raw material liquid in which the juice of the white grapes and the skin of the white grapes are brought into contact with each other and the components contained in the skins of the white grapes are transferred to the juice of the white grapes. May be fermented with.

In the above method for producing wine, the grape is a colored grape or a white grape, and is fermented by the yeast using a raw material liquid containing the juice of the colored grape or the white grape and the peel of the colored grape or the white grape. May be done.

In the present specification, wine refers to liquor obtained by a method including a step of fermenting grape juice. The wine referred to in the present specification includes fruit liquor according to the classification of the Liquor Tax Law and sweet fruit liquor.

According to the present invention, it is possible to provide a method for producing wine with enhanced redness by performing alcoholic fermentation using NYR20 strain in the presence of sulfurous acid.

It is a graph which shows the value of the color density of the wine produced by the method of Example 1 and the wine produced by the method of Comparative Example 1. It is a graph which shows the relationship between the elapsed fermentation days and the amount of carbon dioxide generation in the method of Example 1 and the method of Comparative Example 1, respectively. 3 is a graph showing the color density values of the wine produced by the method according to Example 2, the wine produced by the method according to Example 3, and the wine produced by the method according to Example 4. A graph showing the relationship between the elapsed fermentation days and the amount of carbon dioxide generated in each of the method of Example 2, the method of Example 3, the method of Example 4, the method of Comparative Example 1, and the method of Comparative Example 2. Is. 3 is a graph showing the color density values of the wine produced by the method according to Example 5 and the wine produced by the method according to Comparative Example 3. It is a graph which shows the relationship between the elapsed fermentation days and the amount of carbon dioxide generation in the method of Example 5 and the method of Comparative Example 3, respectively. 6 is a graph showing the color density values of the wine produced by the method according to Example 6 and the wine produced by the method according to Comparative Example 4. 6 is a graph showing the relationship between the elapsed fermentation days and the amount of carbon dioxide generated in each of the method of Example 6 and the method of Comparative Example 4.

Hereinafter, preferred embodiments of the method for producing wine of the present invention will be described.

The present invention is a method for producing wine with enhanced redness using a yeast NYR20 strain (accession number: NITE P-03294) that produces a red pigment, and is described above with respect to a raw material solution containing grape juice and sulfurous acid. It is a method for producing wine including a step of adding yeast and performing fermentation.

The raw material liquid used in the above production method may be one containing grape juice and sulfurous acid. As the raw material liquid containing grape juice, a commercially available grape juice liquid made from colored grapes or white grapes; a liquid containing only colored grapes or fruit juice obtained by crushing white grapes; or colored grapes or Examples thereof include juices obtained by crushing white grapes, peels, flesh and liquids containing one or more of seeds. Sulfurous acid is added to the raw material liquid containing such grape juice for use.

The method for crushing grapes is not particularly limited, and a method for obtaining grape juice may be adopted. For example, the grapes may be crushed, the grapes may be crushed by applying shearing force, or the grapes may be cut into small pieces with a sharp blade. When the grape juice is separated from the peel, pulp, or seeds, the solids may be removed by filtration, or the solids may be precipitated and the supernatant may be recovered as a liquid.

The sulfurous acid to be added is the one commonly used for winemaking. When sulfurous acid is added, it may be added to the raw material solution in the form of sulfite, or sulfurous acid gas may be brought into contact with the raw material solution, or sulfurous acid water may be added by preparing water. Examples of the sulfite salt include potassium pyrosulfite. Examples of the sulfur dioxide gas include sulfur dioxide and the like.

As the yeast, the yeast NYR20 strain (consignment number: NITE P-03294) deposited at the Patented Microorganisms Depositary Center is used. Regarding the culture conditions of the yeast, it grows by culturing under the same conditions as general brewing yeast. When culturing the yeast until the desired number of bacteria is reached, it is preferable to add adenine to the medium. This speeds up the growth of the yeast and makes it possible to reach the desired number of bacteria in a relatively short time.

When the yeast is added to the raw material liquid, it is preferable that the number of bacteria exceeds 1.6 × ¹⁰⁶ cells / mL per 1 mL of the raw material liquid containing grape juice. As a result, the redness imparted to the wine can be further enhanced. Even if the number of bacteria is excessively increased, the effect of enhancing redness tends to be saturated. Therefore, the number of bacteria is preferably 1.0 × ¹⁰⁷ cells / mL or less. On the other hand, the larger the lower limit of the number of bacteria, the more likely the redness is to be enhanced. Therefore, the lower limit of the number of bacteria is more preferably 4.0 × ¹⁰⁶ cells / mL.

When adenine is added to the yeast NYR20 strain in the raw material liquid, the rate of alcohol fermentation by the yeast NYR20 strain is improved. However, the redness imparted to the fermented liquid tends to be lightened. Therefore, when the strength of redness imparted to wine is important, it is better not to add adenine to the raw material liquid. On the other hand, when the speed of fermentation is important, adenine may be added if the amount is small.

The amount of sulfurous acid added is not particularly limited, but for example, sulfurous acid can be added so as to be 50 to 100 mg per 1 L of the fermentation broth containing the raw material liquid and yeast. When a sulfite is blended, it is preferable that the sulfurous acid content is equivalent to 50 to 100 mg / L in consideration of the effective concentration of sulfurous acid. The content of ethanol contained in the fermentation broth varies depending on the raw material and fermentation conditions, but reaches 6.0 to 15.0% by mass. The yeast NYR20 strain can withstand not only the stress caused by sulfite but also the stress of ethanol, undergo alcoholic fermentation, and secrete a red pigment extracellularly.

When white grapes are used as a raw material, fermentation may be carried out with the yeast using a raw material solution obtained by removing the peel of the white grapes from the juice of the white grapes immediately after obtaining the fruit juice from the white grapes. According to this method, although the production method is a method for producing white wine, it is possible to obtain wine having a pale red color such as rosé wine. When the raw material is white grapes, the juice of the white grapes and the skin of the white grapes are brought into contact with each other, and the components contained in the skin of the white grapes are transferred to the juice of the white grapes. It is preferable to carry out fermentation with the yeast. By doing so, it becomes possible to shorten the time required for fermentation and obtain a wine having a red color such as rosé wine.

The grapes are colored grapes or white grapes, and fermentation with the yeast may be carried out using a raw material solution containing colored grapes or white grape juice and colored grapes or white grape skins. According to this method, it is possible to obtain a wine with more redness as compared with a normal wine made from each grape by the NYR20 strain. In this case, the raw material liquid may contain flesh or seeds in addition to the pericarp.

When the yeast is added to a raw material solution containing grape juice and sulfurous acid and fermentation is carried out, sugar or other additives such as brandy may be added.

Examples of colored grapes include black grapes and red grapes. Examples of black grapes include varieties such as Muscat Bailey A and Pinot Noir. Examples of red grapes include varieties such as Koshu and Delaware. Examples of white grapes include varieties such as Chardonnay and Sauvignon Blanc.

Hereinafter, the content of the invention will be described in more detail with reference to examples.

Wines were produced by the methods shown below, and the color density and fermentation process of each wine were examined.

[Example 1]
The yeast NYR20 strain (accession number: NITE P-03294) that produces red color was cultured by the following method. First, inoculum was collected from a glycerol stock of NYR20 strain stored at −80 ° C., and the inoculum was inoculated into a YPD (yeast extract-peptone-dextrose) agar medium having the following composition by a striation method. After culturing the agar medium at 25 ° C. for 5 days, a single colony having a red color was selected from a plurality of colonies appearing on the agar medium and scraped off with a loop loop. YPD medium (yeast extract 10 g; peptone 20 g; dextrose 20 g; agar 20 g; distilled water 1 L)

Put 1 mL of YPAD (yeast extract-peptone-adenin-dextrose) medium in a test tube, incubate at 25 ° C for 24 hours, inoculate 1 mL of this culture solution into a Sakaguchi flask containing 100 mL of YPAD medium, and further inoculate at 25 ° C. The cells were shake-cultured for 48 hours. After culturing, the cells were stored at 4 ° C. for about half a day to a day. YPAD medium (yeast extract 10 g; peptone 20 g; dextrose 20 g; adenine sulfate 0.4 g; distilled water 1 L)

Immediately before the start of fermentation of fruit liquor, a part of the culture solution is diluted 100 times with sterile water, and an inverted phase difference culture microscope (Olympus CK2) is used using a cell counter (Biomedical Science Co., Ltd., product number BMS-OCC01). Under observation, the number of cells (cells / mL) contained in 1 mL of the bacterial solution was measured. Then, the culture solution is weighed so as to have the number of cells to be added to the raw material solution, and the weighed culture solution is centrifuged at 1,700 g for 5 minutes, and the volume of the yeast solution to be added is about 1% of the volume of the sardine. The cells were suspended in sterile water so as to obtain a bacterial solution having a known bacterial count.

According to a conventional method, the juice of black grapes obtained by de-stemming and crushing was put into a fermentation vessel together with the crushed pulp, pericarp and seeds. Muscat Bailey A was used as the black grape. Potassium metabisulfite was added as a sulfite to the raw material solution containing the pericarp, seeds and fruit juice added to the fermentation solution. As for the blending amount of potassium pyrosulfite, 100 mg of potassium pyrosulfite was blended per 1 L of the raw material solution according to a conventional method (blending amount 100 mg / L). The above bacterial solution was inoculated so that the number of cells per 1 mL of the raw material solution containing potassium pyrosulfite was 1.6 × 10 ⁶ to 3.0 × 10 ⁶ (inoculation amount 1.6 × 10 ⁶ to 3). .0 × 10 ⁶ cells / mL). The raw material liquid ingested with NYR20 strain is temperature-controlled to 25 ° C. according to a conventional method, and fermented until the amount of carbon dioxide generated, which will be described later, reaches saturation. When the amount of carbon dioxide generated reaches saturation, fermentation is performed. Was stopped. One week after the start of fermentation, solids containing seeds and pericarp were filtered off. The filtered product was squeezed to produce the wine according to Example 1. When visually observed, the wine was a wine exhibiting a deep red color. In the experiment, the number of samples was set to 3, and the average value of the color density and the average value of the amount of carbon dioxide generated were obtained by the method described later. The same applies to Examples 2 to 6 and Comparative Examples 1 to 4 described later.

[Comparative Example 1]
Wine was produced by the same method as in Example 1 except that the yeast OC-2 strain for red wine brewing, which was sold by NBRC, was changed. When the bacterial solution was added, the conditions were the same as the number of cells added in Example 1 for comparison. When the obtained wine was visually observed, it was a wine having a lighter redness than the wine obtained by the method of Example 1.

[Measurement of color density]
The color densities of the wine obtained by the method of Example 1 and the wine obtained by the method of Comparative Example 1 were analyzed. The color density is the total value of the absorbance value at a wavelength of 420 nm and the absorbance value at a wavelength of 520 nm. There is a correlation between the value of color density and the intensity of red. The higher the color density value, the darker the red. For each wine obtained by the method of Example 1 or Comparative Example 1, the average value of the color densities was calculated. The results are shown in the graph of FIG.

[Analysis of carbon dioxide emissions]
In the method of Example 1 and the method of Comparative Example 1, the relationship between the number of days elapsed after the start of fermentation and the amount of carbon dioxide generated was investigated, with the day of inoculation of yeast as the 0th day. The amount of carbon dioxide generated was measured by measuring the mass of the sardine over time, and the amount of decrease in the mass of the sardine (g) was defined as the amount of carbon dioxide generated (g). The results are shown in FIG. The result of FIG. 2 shows the average value of the three measured values. The amount of carbon dioxide generated correlates with the amount of ethanol produced by fermentation.

As shown in FIG. 1, the method of Example 1 using the NYR20 strain has a higher color density than the method of Comparative Example 1 using the OC-2 strain generally used for winemaking. It can be seen that the value is higher and the redness of the wine is enhanced. In the wine of Example 1, the redness of the wine (A ₄₂₀ + A ₅₂₀ ) was enhanced by 28% as compared with the wine of Comparative Example 1. As shown in FIG. 2, in the method of Comparative Example 1 using the OC-2 strain, the alcoholic fermentation reaches a saturated state about 3 days after the start of fermentation, whereas the example using the NYR20 strain is used. It can be seen that the method 1 requires about 5 times as long as the method of Comparative Example 1 until the alcoholic fermentation reaches a saturated state.

Next, it was investigated how the redness of the produced wine was affected when the number of bacteria added at the time of preparation was increased.

[Example 2]
Wine was produced in the same manner as in Example 1 except that the above bacterial solution was inoculated so that the number of cells per 1 L of the raw material solution containing potassium pyrosulfite was 1.6 × ¹⁰⁶ .

[Example 3]
Wine was produced in the same manner as in Example 1 except that the above bacterial solution was inoculated so that the number of cells per 1 L of the raw material solution containing potassium pyrosulfite was 4.8 × ¹⁰⁶ . When the produced wine was visually observed, it showed a deeper red color as compared with the wine of Example 2.

[Example 4]
Wine was produced in the same manner as in Example 1 except that the above bacterial solution was inoculated so that the number of cells per 1 L of the raw material solution containing potassium pyrosulfite was 1.6 × ¹⁰⁷ . When the produced wine was visually observed, it showed a deeper red color as compared with the wine of Example 2.

[Comparative Example 2]
Wine was produced in the same manner as in Example 1 except that 0.04% by mass of adenine sulfate was added to the raw material solution containing potassium pyrosulfite.

The color densities of the wines produced by the methods of Examples 2 to 4 were measured by the same method as described above. The average value of the color densities was obtained for each of the wines obtained by the methods of Examples 2 to 4. The results are shown in the graph of FIG.

Regarding the methods of Examples 2 to 4 and the method of Comparative Example 2, the relationship between the number of days elapsed after the start of fermentation and the amount of carbon dioxide generated was investigated by the same method as described above. The results are shown in FIG. The result of FIG. 4 shows the average value of the three measured values.

As shown in FIG. 3, in the step of fermenting, the yeast is added to the raw material solution containing grape juice so that the number of bacteria exceeds 1.6 × ¹⁰⁶ cells / mL, and fermentation is performed. In the wines obtained by the methods of Example 3 and Example 4, the method of Example 2 in which the yeast was added so as to be 1.6 × 10 ⁶ cells / mL with respect to the raw material solution containing grape juice. The color density was higher than that of the obtained wine. Among them, it was found that the red color of the wine became the deepest in the method of Example 3 in which the number of bacteria was added three times as much as that of the method of Example 1.

As shown in FIG. 4, in the method of Comparative Example 2 to which adenine was added, alcoholic fermentation proceeded at the same speed as when the yeast OC-2 strain for red wine brewing was used. However, in the wine obtained by the method of Comparative Example 2, the effect of enhancing redness was lost, and the redness was similar to that of the red wine of Comparative Example 1.

Next, it was confirmed by the following method how much redness was imparted when white grapes were used as a raw material.

[Example 5]
Commercially available white grape juice was used as a raw material liquid containing white grape juice and having the white grape skin removed. The juice used was the juice of Chardonnay, a white grape. Potassium metabisulfite was added as a sulfite to this raw material solution. As for the blending amount of potassium pyrosulfite, 100 mg of potassium pyrosulfite was blended per 1 L of the raw material solution according to a conventional method (blending amount 100 mg / L). The above bacterial solution (NYR20 strain) was inoculated so that the number of cells per 1 mL of the raw material solution containing potassium pyrosulfite was 1.6 × 10 ⁶ to 3.0 × 10 ⁶ (inoculation amount 1.6 ×). 10 ⁶ to 3.0 × 10 ⁶ cells / mL). The raw material liquid ingested with NYR20 strain is temperature-controlled to 15 ° C. according to a conventional method, and fermented until the amount of carbon dioxide generated, which will be described later, reaches saturation. When the amount of carbon dioxide generated reaches saturation, fermentation is performed. Was stopped. After the fermentation was stopped, the wine was squeezed to produce the wine according to Example 5. When visually observed, the wine was a wine showing a pale red color like rosé wine.

[Comparative Example 3]
Wine was produced by the same method as in Example 5 except that the yeast used was changed to the yeast W-3 strain for white wine brewing, which was sold by NBRC. When the bacterial solution was added, the conditions were the same as the number of cells added in Example 5 for comparison. When the obtained wine was visually observed, it had the same color as that of commercially available white wine.

The color densities of the wines produced by the methods of Example 5 and Comparative Example 3 were measured by the same method as described above. The average value of the color densities of the wine obtained by the method of Example 5 and the wine obtained by the method of Comparative Example 3 was obtained. The results are shown in the graph of FIG.

Regarding the method of Example 5 and the method of Comparative Example 3, the relationship between the number of days elapsed after the start of fermentation and the amount of carbon dioxide generated was investigated by the same method as described above. The results are shown in FIG. The result of FIG. 6 shows the average value of the three measured values.

As shown in FIG. 5, in the wine produced by the method of Example 5, the red intensity (A ₄₂₀ + A ₅₂₀ ) is higher than that of the wine having the same color as the white wine of Comparative Example 3. It was increased by 63%. However, as shown in FIGS. 2 and 6, the result was that the start-up of fermentation was very slow as compared with the method of Example 1 using black grapes as a raw material.

Next, in the method for producing white wine using the NYR20 strain, the influence of skin contact was confirmed by the following method.

[Example 6]
First, white grapes were de-stemmed and crushed. Shine Muscat was used as the white grape. The crushed mixture containing the flesh, skin, seeds and juice of white grapes obtained by crushing was allowed to stand for 24 hours to transfer the components contained in the skin of white grapes to the juice of white grapes. Then, only the supernatant of the crushed mixture was recovered and used as a raw material liquid. In the subsequent process, the wine according to Example 6 was produced in the same manner as in Example 5 above. When visually observed, the wine was a wine showing a pale red color like rosé wine.

[Comparative Example 4]
Wine was produced in the same manner as in Example 6 except that the yeast to be inoculated was changed to a strain (OC-2) generally used for winemaking. When visually observed, the wine had a color similar to that of commercially available white wine.

The color densities of the wines produced by the methods of Example 6 and Comparative Example 4 were measured by the same method as described above. The average value of the color densities was obtained for each of the wines obtained by the method of Example 6 and the method of Comparative Example 4. The results are shown in the graph of FIG.

Regarding the method of Example 6 and the method of Comparative Example 4, the relationship between the number of days elapsed after the start of fermentation and the amount of carbon dioxide generated was investigated by the same method as described above. The results are shown in FIG. The result of FIG. 8 shows the average value of the three measured values.

As shown in FIG. 7, in the wine produced by the method of Example 6, the intensity of red color (A ₄₂₀ + A ₅₂₀ ) is higher than that of the wine having the same color as the white wine of Comparative Example 4. It was enhanced by 74%. Further, in the method of Example 6, by performing skin contact, as shown in FIG. 8, the time required to complete the fermentation was significantly shortened.

It is known that a part of sulfurous acid added to wine binds to organic matter in fruit juice to form bound sulfurous acid. It is the remaining free sulfurous acid that exhibits bactericidal activity. It is considered that the reason why the fermentation rate in the method of Example 5 was significantly delayed as compared with the method of Example 1 was that the amount of free sulfurous acid increased due to the small amount of organic matter in the fruit juice. In the method of Example 6, it is considered that the fermentation inhibitory action on the NYR20 strain was weakened by changing the free sulfurous acid in the fruit juice to the bound type.

According to the method for producing wine of the present invention, the redness of wine can be enhanced. For example, due to the effects of global warming, the temperature difference between day and night is becoming smaller. The coloration of grapes is affected by the temperature difference between day and night, but the problem is insufficient coloring of grapes caused by the small temperature difference. By using the method of the present invention, the redness of wine can be enhanced, so that poorly colored grapes can also be used as a raw material for wine. Further, with the conventional method, only white wine can be produced from white grapes such as Chardonnay and Sauvignon Blanc, but by using the method of the present invention, rosé wine can be brewed.

Claims (5)

It is a method for producing wine with enhanced redness using yeast NYR20 strain (accession number: NITE P-03294) that produces red pigment.
A method for producing wine, which comprises a step of adding the yeast to a raw material liquid containing grape juice and sulfurous acid to ferment. The step of fermenting is according to claim 1, which is a step of adding the yeast so that the number of bacteria exceeds 1.6 × 10 ⁶ cells / mL with respect to the raw material liquid containing grape juice and fermenting. How to make wine. The grapes are white grapes,
The method for producing wine according to claim 1 or 2, wherein immediately after obtaining fruit juice from white grapes, fermentation with the yeast is carried out using a raw material solution obtained by removing the peel of white grapes from the fruit juice of white grapes. The grapes are white grapes,
Contact the juice of white grapes with the peel of white grapes,
The method for producing wine according to claim 1 or 2, wherein the component contained in the peel of white grapes is fermented with the yeast using a raw material liquid obtained by transferring the juice of white grapes to the juice of white grapes. The grapes are colored grapes or white grapes,
The method for producing wine according to claim 1 or 2, wherein fermentation is carried out with the yeast using a raw material liquid containing colored grape or white grape juice and colored grape or white grape skin.

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