KR100886954B1 - Novel fermentation strain s. cerevisiae ij 850 isolated from geubong grape and method of manufacturing wine using it, and wine manufactured by method thereof - Google Patents

Abstract

The present invention is a new fermented strain Saccharomyces cerebase ( Saccharomyces isolated from grape grapes) cerevisiae ) IJ850 and a method of making wine using the same, and wine produced by the method, which has a high fermentation rate during the wine making process using grape grapes and has excellent alcohol fermentation ability and low risk of germs. Saccharomyces cerevisiae ) IJ850 (KACC 93054P) fermented strains can reduce fermentation cost and fermentation failure rate, and improve the quantity and quality of domestic wine production. To this end, the present invention is a novel fermented strain Saccharomyces cerevisiae isolated from giant grapes ( Saccharomyces cerevisiae ) IJ850 (KACC 93054P), a wine making method using the same and wine produced by the method.

Geobong grape, Saccharomyces cerevisiae, IJ850 (KACC 93054P), fermentation, identification, separation, physical and chemical comparative analysis

Description

NOVEL FERMENTATION STRAIN S. CEREVISIAE IJ 850 ISOLATED FROM GEUBONG GRAPE AND METHOD OF MANUFACTURING WINE USING IT , AND WINE MANUFACTURED BY METHOD THEREOF}

1 is a novel alcohol fermented strain Saccharomyces cerevises according to a preferred embodiment of the present invention ( Saccharomyces cerevisiae ) Process flow chart to explain the screening process of IJ850 (KACC 93054P)

Figure 2 is a graph showing the NCBI blast results for the purpose of identification of novel fermentation strains according to the present invention

Figure 3 is a graph showing the change in sugar (° Brix) during the fermentation process of the strain to the giant grape grapes according to a preferred embodiment of the present invention

Figure 4 is a graph showing the change in alcohol concentration (%) during the fermentation process of the strain to giant grape grapes according to a preferred embodiment of the present invention

Figure 5 is a graph showing the change in total acidity (g / L tartaric acid) during the fermentation process of the strain to giant grape grapes according to a preferred embodiment of the present invention

Figure 6 is a graph showing the change in hydrogen ion concentration (pH) during the fermentation process of the strain to giant grape grapes according to a preferred embodiment of the present invention

Figure 7 is a graph showing the content of phenolic components (mg / L) after fermentation of strains to giant grapes according to a preferred embodiment of the present invention

The present invention is a new fermented strain Saccharomyces cerebase ( Saccharomyces isolated from grape grapes) cerevisiae ) IJ850 and the wine production method using the same and the wine produced by the method, more specifically, the fermentation ability of the yeast used to make wine from grape grapes relatively saccharo than the commercial strains Saccharomyces cerevisiae IJ850 (KACC 93054P), and a wine manufacturing method using the same and wine produced by the method.

 Fermentation is a phenomenon in which microorganisms decompose organic acids of relatively large molecules, and the energy generated by them is used as a driving force for living organisms. This action is a phenomenon that can be seen in all the flora and fauna, the fungus is particularly strong in the action there is a case to use it in human life. All these fungi are called fermented bacteria (yeast). Fermentation bacteria produce various substances in the body, including vitamins, nucleic acids, minerals, hormones and fatty acids. In addition, humans produce eight essential amino acids that can't be synthesized in the body, so eating a lot of fermented foods containing yeast is healthy.

Among fermented fungi, the ability to break down sugar to produce alcohol has been used for brewing since ancient times. Brewer's yeast ( Saccharomyces cerevisiae ) was recorded in 1883 as a species that causes beer fermentation. The yeast used to brew the sake was described in 1897 as another species called Saccharomycs sake , but recent studies have shown it to be homologous to brewer's yeast. Commonly used baker's yeast is also known to belong to this species. In particular, wine yeast making Saccharomyces cerevisiae ( Saccharomyces cerlipsiae ), Saccharomyces ellipsoideus ( Saccharomyces ellipsoideus ) is mainly used Saccharomyces ellipsoidus . These are attached to the fruit of the grapes.

In recent years, with the growing interest in wine with well-being, Korea's production is growing very fast by 20 ~ 30% every year, but it still depends on imports, and its imports are gradually increasing. This is because the cultivation of brewing grapes is not suitable due to the climatic conditions of our country, so there are many difficulties in wine production. In Korea, many studies on wine production are constantly being made, and as the most familiar edible grapes in Korea, many varieties of Vitis labruscana B and Vitis L. are produced. However, the demand for domestically produced wine is not only incomplete compared to imported wine, but also needs to be improved in quality.

Conventionally, the quality of wine is considered to be different only by wine juice, which is a raw material, and active research has not been conducted on the research and improvement of the manufacturing process. However, as it has recently been found that wine yeast plays an important role in the formation of the taste of wine and the properties of the wine produced by the wine fermentation strains used vary greatly, the means for breeding of wine yeast, for example, search, separation Interest in genetic engineering techniques, such as mutation, conjugation, transformation, and cell fusion. This is also the result of research of the prior art.

Korean Patent Publication No. 1995-0004007 also discloses a novel brewing yeast using a cell fusion method and a method of preparing wine using the same. This relates to fusion cells SW1 and SW6 prepared by cell fusion of wine yeast Saccharomyces cerbize 1032 and 1064 and sake chemistry yeast Saccharomyces cerbize 1137.

Korean Patent Publication No. 1991-0004364 discloses ethanol at a high temperature of 35 ° C. or higher using Saccharomyces cerevisiae JY 10035, a new microorganism isolated from UV-transferred yeast, which is the most commonly used for producing fermented alcohols made from starch. The method of preparation is shown. In addition, Korean Patent Publication No. 1999-0038446 discloses Saccharomyces cerevisiae YJK 20, a strain having excellent alcohol fermenting power from pulp and stalks of spoiled persimmons.

In view of the fact that the domestic grape cultivation varieties are limited to edible grapes, the inventors of the present invention also have to study the fermented strains and manufacturing process prior to improving the quality of domestic wine.

Geobong grapes were named in 1937 in Japan by Mr. Oi and the centennial grape variety of the Savior quadrant, which is a tetracot variety of Campbell's Early. It is a tetraploid mutant breeded by breeding and growing in Korea. Currently, Anseong province in Gyeonggi-do is the main producing region of Geobong grapes. Tetraploid refers to the doubling of chromosomes of common organisms through chemical treatment. In other words, there are 38 chromosomes in Genuine grapes or 40 chromosomes in Genus Muscotia, but there are 76 or 80 chromosomes in tetraploid grapes. The characteristic of these four-ply plants is that fruits, leaves, stems, etc. are larger than normal two-ply plants.

In the Anseong area, where grapes are grown under strict quality control called 'perfect grapes', most of the production of raw and processed wines is being demonstrated by some farms or union units. The grape farming cooperative located in Seoun-myeon, Anseong produces wine distillation with complete wine processing facilities, but it is difficult to produce high quality and standardized products by combining scientific theory and technology. This is impossible and sales are not working properly. Thus, the inventors of the present invention tried to develop a strain suitable for the grape varieties varieties by producing wine using the wild strain isolated from the grape grapes as part of the improvement of the quality of domestic wine and analyzing its fermentation characteristics. There is no known fermented strain suitable for making wine from grapes. Since most of the fermentation strains used in domestic wine use commercial strains developed in foreign countries, it is considered that the development of fermented strains suitable for the Korean edible grape varieties Geobong grapes is very important.

Accordingly, the present invention has been made to solve the above problems, and a first object of the present invention is to provide a new fermentation strain having a high fermentation capacity suitable for giant grape varieties and excellent alcohol production ability and a low risk of bacterial growth. .

In addition, a second object of the present invention is to provide a new fermentation strain that can reduce the fermentation cost and fermentation failure rate, and contribute to the quality improvement of grape wine grapes.

In addition, a third object of the present invention is to provide a new fermented strain that can replace a foreign commercial strain that is widely used as a fermentation strain when making wine from grape grapes.

In addition, a fourth object of the present invention is to provide a wine production method using a new fermented strain isolated from giant grapes and wine produced by the method.

The new fermented strain Saccharomyces cerevisiae jae 850 (IJ850) and the wine production method using the same and the wine produced by the method is isolated from the giant grapes according to the present invention for achieving the above technical problem, the fermentation rate is fast alcohol producing ability is excellent, and various bacteria the risk that using a grape in a new fermentation strain Saccharomyces isolated from grapes in saccharide as MRS celebrity non-zero (Saccharomyces cerevisiae) IJ850 and, wine fermentation strains MRS celebrity non-zero (Saccharomyces cerevisiae) IJ850, characterized in that the low Wine production method characterized in that using wine and Saccharomyces cerevisiae ( Saccharomyces cerevisiae) IJ850 (microbial accession number: KACC 93054P, depository institution: Institute of Agricultural Biotechnology, date of deposit: May 30, 2007) It consists of.

Saccharomyces cerevisiae IJ850 (KACC 93054P), a new fermented strain isolated from giant grapes according to the present invention, is isolated from giant grapes and is suitable for giant grapes having a very high alcohol production rate and low risk of germs when fermented. As a fermentation strain, fermentation cost and fermentation failure rate are reduced by using this, which can contribute to the quantitative and qualitative improvement of domestic wine production.

The procedures for IJ850 isolation, identification and physicochemical analysis are as follows.

 The pulverized giant grape grapes were sampled once a day for 3 days through the fermentation process at 30 ° C., and 100ul was selected as a selective medium plate (pH 4.5, yeast extract 1%, peptone 2%). , Spread by dextrose (2%, Agar 1.5%) and confirmed by 3 days. Dominant spawns are selected that show over 80% of each sample after 48 hours of incubation.

Next, DNA of Dominant spawn was extracted and PCR homologous reaction of D1 / D2 region was performed to compare homology of 26SrDNA fragments. S accharomyces cerevisiae .

Then, IJ850 is inoculated into the culture medium of the selective medium, followed by centrifugation after 48 hours of incubation, and cell culture, and used as a starter for fermentation of IJ850.

Then, IJ850 and commercial strains are inoculated into each of the shredded giant grape grapes, and samples are taken from each fermentation vessel every day during fermentation for 16 days from the start of fermentation. After completion of fermentation, bentonite is added to undergo primary and secondary sedimentation. After 3 weeks, feed through the filtration process.

Hereinafter, with reference to the accompanying drawings will be described in detail a preferred embodiment of the present invention.

1 is a process flow chart illustrating a screening process of a novel fermented strain Saccharomyces cerevisiae IJ850 (KACC 93054P) according to a preferred embodiment of the present invention.

Figure 2 is a graph showing the NCBI blast results confirmed the species name for the purpose of identifying a new fermentation strain according to a preferred embodiment of the present invention.

Figure 3 is a graph showing the change in sugar (° Brix) during the fermentation process of the strain to giant grape grapes according to a preferred embodiment of the present invention.

Figure 4 is a graph showing the change in alcohol (%) during the fermentation process of the strain to giant grape grapes according to a preferred embodiment of the present invention.

Figure 5 is a graph showing the change in total acidity (g / L tartaric acid) during the strain inoculation fermentation process to giant grapes according to a preferred embodiment of the present invention.

Figure 6 is a graph showing the change in hydrogen ion concentration (pH) during the fermentation process of the strain to the giant grape grapes according to a preferred embodiment of the present invention.

Figure 7 is a graph showing the content of phenolic components (mg / L) after fermentation of strains to giant grapes according to a preferred embodiment of the present invention.

The grape used in the above experiment is a giant grape harvested in Anseong, Gyeonggi-do, and the sugar added for wine fermentation is glucose (non-hydrogenated glucose, Q1). In addition, the fermented strains used were IJ850 strain (KACC 93054P) isolated from giant grapes and seven commercial strains used for wine fermentation. These strains are shown in Table 1 below.

Strains Trade name species RSMR RED STAR Montrachet Red S. cerevisiae RSPC RED STAR Premier Cuvve S. bayanus CBY Coopery Brewing Yeast S. cerevisiae LBB LEVOCLL BB (Italy) S. bayanus LQB-EG Muntons Gold old english bitter LQB EG S. cerevisiae, ect. EC1118 LALVIN EC-1118 S. cerevisiae CY Cultivation yeast S. cerevisiae, ect. IJ850 Isolated from Anseong Geubong wild yeast S. cerevisiae

And new fermented strain Saccharomyces ( Saccharomyces) cerevisiae ) IJ850 is currently deposited (KACC 93054P) at the Korea Agricultural Microbial Resources Center (225 Seodun-dong, Gwon-gu, Suwon-si, Gyeonggi-do).

Example 1 Screening of Strains

Experimental Example 1 Isolation and Identification of Alcohol Fermentation Strains

At 30 ° C fermentation conditions, samples were taken once a day for three days using the crushed giant grapes, and 100ul was selected as a selective medium plate (pH 4.5, yeast extract 1%, peptone). 2%, Dextrose 2%, Agar 1.5%) was spread (spreading). After 48 hours of incubation for each sample, the dominant spawn was selected. DNA was extracted from the cells of the selected dominant spawn IJ850 and then subjected to PCR-Polymerase Chain Reaction of the D1 / D2 region of chromosomal DNA. In addition, NCBI blast comparison of 26SrDNA fragments revealed that the species of IJ850 was saccharomyces cerevisiae. I could confirm it. The results are shown in the table shown in FIG.

Then, in order to use as a starter for fermentation of IJ850 (KACC 93054P), IJ850 was inoculated into 500 ml of selective medium culture and centrifuged at 12,000 rpm for 48 hours, followed by cell culture.

Experimental Example 2 Fermentation and Physicochemical Analysis Using IJ850

First, 180 kg of giant grapes were washed twice with clean water, the stems were removed, and then the shells were removed. Then, 20 kilograms of grape kernels were collected in a 20 liter fermenter and then crushed. Then, in order to ensure that the sugar content of the grape juice is 28 ° Brix, 18 ° Brix grape juice must be filled with 10 ° Brix short, 18 ° Brix has a specific gravity of 1.072 and sugar is 195 g / L. Since 28 ° Brix has a specific gravity of 1.117 and a sugar content of 314 g / L, 314-195 = 119, that is, 119 grams of glucose was added per liter.

Then, to prevent browning and contamination, potassium metabisulfite (K ₂ S ₂ O ₅₎ was initially added at a concentration of 50 ppm, that is, 50 milligrams of sulfite salt per liter of grape juice. Then, it was left at room temperature for 1 hour. Then, 48 hours after the inoculation of 500 milliliters of IJ850 strain, the starter and 7 commercial strains cultured with the cells were inoculated into fermenters.

During the fermentation and maturation for 16 days from the start of fermentation of the seven starter commercial strains, samples were taken daily from each fermentation vessel and subjected to physicochemical analysis.

1. Changes in sugar (° Brix) during wine fermentation

The sugar content of wine during fermentation was measured using a hand refractometer (ATAGO, N-1α, Japan) at room temperature. The results are shown in the graph shown in FIG.

All strains started to ferment after raising the sugar content from 17 ° Brix to 28 ° Brix. As the sugar content decreased rapidly from 8 to 15 ° Brix until 8 days of fermentation, it was found that the wines of 8 strains were actively fermented and gradually decreased after 13 days of fermentation. On the second day of fermentation, the sugar content of IJ850 was the lowest as 23.8 ° Brix. In addition, the results of the fermentation rate up to 4 days, the strain RSMR, CBY, LBB and IJ850 wine of 11 ° Brix compared to other sugars range from 19 to 22 ° Brix range was faster fermentation rate. Through this, IJ850 consumed glucose faster than other strains, demonstrating that the initial growth capacity is fast and the fermentation force is strong.

2. Changes in alcohol content (%) during wine fermentation

Alcohol content was measured using a spirit meter after adjusting the amount to 100 milliliters at 15 ℃ after the first distillation. The results are shown in the graph shown in FIG.

In proportion to the amount of sugar changed, fermentation was actively carried out up to 8 days of fermentation and after 13 days, fermentation was completed. 11.6% of strains RSMR, CBY, LBB and 12% of IJ850 in the first 6 days of fermentation, relative to strains RSPC (7.2%), LQB-EQ (7%), EC1118 (8.4%) and CY (6%) High alcohol content. Also, when the alcohol content at the end of fermentation was 15.6% of strain RSMR, 14.8% of CBY, 15.6% of LBB, and 16.8% of IJ850, strain IJ850 produced faster final alcohol and produced faster than other commercial strains. It can be seen that the alcohol content is also the highest.

3. Changes in g / L tartaric acid during wine fermentation

Total acidity was measured as a comparative amount of tartaric acid. Five drops of a phenolphthalein solution (1% in methanol) was added to a 10 milliliter sample and titrated with 0.1 N sodium hydroxide (NaOH) using a burette on a stirrer. The amount of sodium hydroxide in which the sample remained pink for about 15 seconds was measured. The results are shown in the graph shown in FIG.

In general, the total acidity (g / L tartaric acid) is inversely related to the sugar content. At the beginning of fermentation, the total acidity was 3.52, but it increased greatly from 6.62 to 8.58 during the first two days after fermentation, and steadily increased from 7.4 to 8.4 until 6 days later. Could see. IJ850 also proved to be a moderate sour strain by showing a moderate graph of all strains not too high or too low for acidity.

4. Changes in pH during the fermentation of wine

Hydrogen ion concentration (pH) was measured using a hydrogen ion concentration meter (pH meter) (ORION, 420A, USA). The results are shown in the graph shown in FIG.

Wine is preferably 3.3 at pH 3.2. If the pH is 3.6 or higher, there is a high possibility of the occurrence of various germs during storage. On the contrary, if the pH is 3.2 or lower, it is too acidic. At the beginning of fermentation, all wines had a pH of 3.49, but on the second day of fermentation, all strains except EC1118 and CY had dropped to below pH 3.3. If the high pH at the beginning of fermentation, there is a risk of the occurrence of various bacteria, but IJ850 showed a low pH of 3.24 and showed a medium position after the completion of fermentation.

5. Phenol Content after Fermentation

Total phenolic content was used by Swain and Hillis. That is, after reacting with a Folin-Ciocalteu reagent (Sigma-Aldrich, USA), it was measured at 750 nm using a spectrophotometer (Mecasys, OPTIZEN 2120UV, Korea). The results are shown in the graph shown in FIG.

The total content of phenolic components ranged from 11 to 16.87 mg / L. The phenol component, which is a functional compound, showed an intermediate position of 13.74 mg / L compared with other strains, and thus, it was confirmed that the strain was suitable for the production of functional substances.

6. Determination of Color of Wine after Fermentation

  After fermentation, the color of the wine was measured five times using a color color meter (Color Techno System Corp., JC801, Japan), and then the average value L * (brightness lightness), a * (redness redness), Chromaticity is indicated by the b * (yellowness) value. The results are shown in Table 2 below.

Strain L * a * b * RSMR 99.5 -0.3 2.1 RSPC 99.4 -0.2 2.3 CBY 99.5 -0.5 2.5 LBB 99.4 -0.3 2.5 LQB-EG 99.4 -0.5 2.6 EC1118 99.4 -0.2 2.4 CY 99.3 -0.4 3.1 IJ850 99.7 -0.5 2.1

The degree of yellow color (b *) did not show a big difference compared to other strains, so there is no possibility of producing a cloudy or brown-colored wine. IJ850 (KACC 93054P) is -0.5 on the basis of the redness level of 0 to compensate for the shortcomings of wines made with light red grapes compared to other strains (-0.2 to -0.5). I can say that.

As in the present invention, the fermented strain Saccharomyces cerevisiae IJ850 (KACC 93054P) is a fermentation rate is high when the wine is made from grape grapes as a fast fermentation rate accordingly has excellent alcohol production ability and low risk of germs It is a very suitable strain for wine production using grapes.

Those skilled in the art should understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. The scope of the invention is indicated by the following claims rather than the above description, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the invention. do.

As described above, according to the present invention, by using the new fermented strain Saccharomyces cerevisiae IJ850 (KACC 93054P) isolated from the edible varieties of Korean edible varieties, wine can be produced pure domestically It works.

In addition, fermentation cost and fermentation failure rate are reduced by using Saccharomyces cerevisiae IJ850 (KACC 93054P) strain, which has a high fermentation rate and high alcohol production ability and low risk of germs. It works.

In addition, by reducing the import of foreign commercial strains that depend on the fermentation strain of domestic wine production, there is an effect that can contribute to the domestic wine industry economy.

Claims (3)

Saccharomyces cerevisiae IJ850 (KACC 93054P), a new fermentation strain with excellent alcohol production ability isolated from grape grapes Wine production method using Saccharomyces cerevisiae IJ850 (KACC 93054P) as a wine fermentation strain of grapes grapes Saccharomyces Isolated from Giant Grapes cerevisiae ) Wines made using IJ850 (KACC 93054P)

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