KR101541518B1 - Saccharomyces cerevisiae GBY1 Yeast for Fermentation of Wine with Berries Fruits and Manufacturing Method of Berries Fruits Wine Using Fermentation - Google Patents

Abstract

The present invention relates to a method for producing a berries fruit wine using native fermentation yeast. More specifically, the fruit is thawed and crushed, saccharides are added to adjust the sugar content, and yeast ( Saccharomyces a fermentation step of inoculating and fermenting the microorganism deposited with the microorganism preservation center of S. cerevisiae GBY1, KCCM11417P, 2013. 5.22), fermenting the fermentation after completion of the fermentation, ≪ / RTI >
The berries of the present invention include brambles, olives and blueberries.

Description

[0001] Description [0002] Saccharomyces cerevisiae GBY1 Yeast for Fermentation of Wine with Berries Fruits and Manufacturing Method [0003] Berries Fruits Wine Using Fermentation [

The present invention relates to a yeast for fermentation of berries and a fermentation method of a fruit wine using the yeast. More specifically, the fruit is thawed and crushed, saccharides are added to adjust the sugar content, and yeast ( Saccharomyces cerevisiae GBY1, Korean Microorganism Preservation Center Accession No .: KCCM11417P, 2013. 5.22 deposit) and fermenting after fermentation is completed, followed by juicing and freezing filtration, followed by aging in a cold dark place.

In the present invention, berries fruit fruits include Rubus, Audi and Blueberries.

Rubus occidentalis ) is a deciduous shrub of Rosaceae and is grown in the southern and central parts of Korea. It ripens in mid-June to July-August, ripens in round and red color and then matures into reddish-brown color with sweetness, sourness and unique fragrance It contains phenol compounds such as kaemferol, sanguinin H-5, 3-Ο-β-D-glucuronide, quercetin, ferulic acid, coumaric acid, caffeic acid, rutin , luteolin, etc. have been reported. Studies on bokbunja have been reported on tannin and flavonoids compounds from leaves and stems. Bokbunja is used for edible and medicinal purposes. Fruits are mainly used in food, beverage and alcoholic products. In Oriental medicine and medicinal berry, fruit protects the liver, clears the eyes, darkens the hair and treats infertility. It is recorded. There are various studies on the components and physiological activity of the bokbunja fruit. The functionalities of the bokbunja fruit known to date include antioxidant, antibacterial, antiallergic, anticancer, antiinflammatory, hormone control, skin whitening, obesity, Blood sugar, blood pressure, cholesterol).

Mulberry (Morus bombycis Koidz) is a mulberry fruit, similar to a strawberry-like liquid. Its nucleus hangs only on the rocks. It is initially blue, but gradually reddish. When ripened, it changes from purple to dark purple. The pigment belongs to the cyanidin family as an anthocyanin component. Audi has been widely recognized for its effectiveness in health and medicine, and in Korea and China, it has been treated as a very precious drink because it is called mulberry wine. Audi has a sour taste, so it is made of fine suture. Audis helps blood circulation and activates metabolism, which is good for hypotension, poor circulation and insomnia. The juice of the Odi makes about 80% of the juice, which contains more than 10% of the sugar, which is strong in sweetness.

The blueberry is called a bilberry and has anthocyanin color, blue sugar, sweet and sour sugar, viscous pectin, and a scent of fragrance. Blueberries contain 4.5 grams of dietary fiber per 100 grams and are rich in calcium, iron, and manganese. Blueberries have long been used as food by Indians in North America, and fruit and leaf extracts have been used in the treatment of scurvy, diabetes, and urinary disorders. Blueberries contain anthocyanins to keep your eyes healthy and contain a lot of antioxidants and dietary fiber, which helps to maintain health and youth. Candy, gum, jam, and drinks are produced worldwide as blueberry related products. In recent years, functional foods and medicines have been actively promoted, focusing on the functionality of eyes. The blueberries are mainly produced in the USA, Canada, Japan, Germany, New Zealand, etc. Recently, however, the fruits have been cultivated in Korea and the development of related products has been steadily developed.

Korean Patent Laid-Open Publication No. 2005-0066429 discloses a method for preparing a bokbunja, comprising the steps of: a) crushing bokbunja and then adding sugar to adjust the sugar content of the bokbunja to 25-35brix; b) 10 to 50 ppm of sulfurous acid and dried yeast for wine are added to the brittle fracture with controlled sugar content in step a), and fermentation is carried out at 20-30 ° C for 7-8 days; c) filtering the fermentation product obtained in step b); d) adding alcohol and water to the fermentation broth obtained in step c) to form a final alcohol concentration of 14-19% (v / v); And e) adding 1 to 5% (w / v) or 10 to 30% (v / v) of concentrated juice to the syrup produced in step d). Korean Patent Registration No. 664905 (method of producing grape berry wine) is a method of (1) harvesting ripe brambles in a state where the harvest time of grapes and brambles do not match, (2) defrosting the brambles at the grape harvesting time, crushing them with a crusher, and crushing the harvested grapes; (3) controlling the mixing of the crushed fruit juice, the sugar content and the acidity; (4) To add 0.02-0.04 g / l of pectinolytic enzyme to the juice of the above (3), 0.09-0.2 g / l of potassium metabisulfite is added to the juice to sterilize the juice step; (5) Inoculating yeast to a part of the juice prepared in the step (4) to produce a beverage; (6) adding alcohol to the mixed juice obtained in (4) by the step (5); (7) purifying the fermented beverage according to the above (6) by adding clarifying agent of 0.2 to 1 g / l of bentonite and 0.01 to 0.3 g / l of gelatin to clarify; (8) A method in which the fermentation product of (7) is filtered after filtration. Korean Patent Registration No. 759267 (Method of manufacturing bokbunjugu) is a process for extracting a pigment and a physiologically active substance contained in bokbunja fruit by treating frozen bokbunja fruit with hot water at 80 ~ 90 ℃. b) adding sugar to the product obtained in step a) to adjust sugar content and adding yeast to ferment; c) filtering the fermentation product obtained in step b); d) post-fermenting the fermentation broth obtained in step c); e) aging the fermentation broth obtained in step d); And f) adding the oak slices to the brambles obtained in step e). Korean Patent Registration No. 10-650193 (a method of manufacturing high quality bokbunja) is a process of treating pectin degrading enzyme in a bokbunja juice crushing liquid and adding a sulfite to a bokbunja juice to inhibit the propagation of harmful bacteria, ( S. cerevisiae KCCM 11352 and 12224) obtained by purely isolating yeast from a traditional Chinese cabbage, marjoram or meju with good flavor and taste, Preparing a starter by inoculation, and inoculating and culturing the starter into the pretreated bokbunja and juice to produce a bokbunja. In addition, there are Korean Patent Registration Nos. 10-0628336 and 10-0759267 as manufacturing methods of bokbunjangju.

However, these prior arts have different technical constructions from those of the present invention.

The currently distributed fruit is mixed with sugar, fruit juice is mixed with sugar, and the fruit is matured, or sugar is added to the fruit of the fruit, and the mixture is fermented for 27 days. Then, the fruit is added to the fruit juice and fermented for 4060 days. Therefore, a method of extracting various components contained in fruit can be referred to as alcohol-enhanced fermentation rather than fermentation. As described above, the alcohol-fortified buppies are leached for a long period of time, resulting in a large amount of tannin eluted from the seeds, resulting in a strong sour taste, lowering the color of bokbunja due to incomplete fermentation management and incomplete fermentation. In addition, it is difficult to standardize the quality, and the taste is not advanced, and the market is gradually disappearing. Odin and blueberries are provided only as fruits and juices, and products are not diversified.

Among berry fruit berries, especially brewers, it is urgent to improve the quality of brewer's brewers in the form of fermented brewers rather than alcohol-fortified brewers, under excessive competition to supply them at lower price than quality, as producers and factories are nationwide scarce. It is also important to select strains to provide a uniform and unique taste to consumers. Therefore, the present invention relates to a fermentation yeast ( Saccharomyces) which excelles ethanol fermentation of berries fruit (bokbunja, audi, blueberry) cerevisiae GBY1, Korean Microorganism Conservation Center Accession No .: KCCM11417P) was isolated and the fermentation of berries fruit was carried out.

The present invention relates to a method for fermenting fruit by thawing and crushing fruit, adding saccharide to adjust sugar content, fermenting yeast ( Saccharomyces cerevisiae GBY1, Deposit number KCCM 11417P, deposited at Korea Microorganism Conservation Center, 2013. 5.22 deposit) Followed by freezing filtration and aging in a cold dark place.

The present invention can provide a high quality berries fruit wine because it is excellent in ethanol-producing ability and is used for isolating yeast that harmonizes taste and aroma of berries fruit wine with berries.

Figure 1 is a colony of the strain of the present invention ( S. cerevisiae GBY1, KCCM11417P).
Figures 2a and 2b are the nucleotide sequences of molecular biology identification.
Fig. 3 shows the growth curve of the strain of the present invention.
4 shows the alcohol resistance of the strain of the present invention.
Fig. 5 shows the resistance of the strain of the present invention.
6 shows the results of bacterium alcohol fermentation using GBY1 strain of the present invention.
Fig. 7 shows the result of the fermentation of the aldehyde using GBY1 strain of the present invention.
Fig. 8 shows the result of blueberry alcohol fermentation using GBY1 strain of the present invention.

The present invention adjusts the sugar content to thaw the fruit and disrupted by the addition of sugars and yeast (Saccharomyces cerevisiae GBY1, Korean Microorganism Preservation Center Accession No .: KCCM11417P, 2013. 5.22 deposit) and fermenting after fermentation is completed, followed by juicing and freezing filtration, followed by aging in a cold dark place.

1) Strain isolation

Saccharomyces cerevisiae isolated from Kochongsan Audi fermentation broth and strain GBY1 ( Saccharomyces cerevisiae GBY1, Korean Microorganism Preservation Center, Accession No .: KCCM11417P) was cultured in YPD medium at 25 ° C for 48 hours. When the colonies of FIG. 1 were streaked and observed, the colonies were round in shape and spherical in microscopic observation. The strain was relatively large and ivory, smooth on the surface, convex on top, and round in shape.

Biochemical patterns (- + ------ + - + - + - + + - + -) digitized using API 20 C AUX (yeast identification kit), GOOD IDENTIFICATION , But the strains did not show Saccharomyces cerevisiae 1,% ID 97.9, T index 0.86, opposite biochemical characteristics (MDG), catalase test (positive reaction) and hemolysis test. In the biochemical pattern, '+' means a positive reaction, '-' means a negative reaction, and the first '-' of the above symbols is a result of a control group.

The nucleotide sequence of 18S rRNA analysis was as shown in Fig. 2, using ITS1 (CC GTA GGT GAA CCT GCG G) and ITS4 (TCC TCC GCT TAT TGA TAT GC) primers for molecular biology identification. The growth curve was as shown in Fig. 3 when the absorbance at 600 nm was measured at intervals of 6 hours.

2) Alcohol tolerance

The culture broth was inoculated 1% (v / v) in 50 mL of the culture medium using YPD medium containing 0, 5, 10 and 15% of the initial alcohol, and cultured at 37 ° C and 160 rpm for 24, 48 and 72 hours. The absorbance was measured and shown in Fig. Overall, inhibition of yeast growth was found in the medium containing alcohol. In the 5% alcohol-containing medium, about 13% of the growth inhibition was not observed, and the growth inhibition in the alcohol-containing medium of 10% or more was observed to be insufficient. Therefore, the strain GBY1 appears to be resistant to alcohol. The results are shown in Fig.

3) My herbicide

The culture broth was inoculated 1% (v / v) in 50 mL of the culture medium using YPD medium containing 0, 10, 20 and 30% initial glucose concentration and cultured at 37 ° C and 160 rpm for 24, 48 and 72 hours, And the results are shown in Fig. Overall, the growth of strains tended to increase in the glucose addition section. 30% glucose was added to the medium. The results are shown in Fig.

4) Alcohol fermentation

After sacrificing berries berry, oak and blueberry fruit, sugar is added to 20% of the fruit, yeast is inoculated at 1 × 10 ⁹ CFU / mL per 1 kg of fruit, fermented at 25 ° C, The change was confirmed, and the results are shown in Fig.

<Example 1> Bokbunja alcohol fermentation using GBY1

When 20% sugar of raw material was added to the berry fruit, the sugar content was 30.7 ° Bx. Adding enzyme viscozyme 600ppm, and decreased for the strain of the present invention to 10 ⁹ CFU / mL / fruit 1kg added to 25 ℃ fermentation, the 25.5 ° Bx reached the first day when the check quality change interval 1 il produced alcohol 2.73% . The sugar content was steadily decreased, and the alcohol was formed and the alcohol was formed at about 15.3% on the sixth day. When the same fermentation was carried out with the commercial strain Fermivin, alcohol was produced starting from the initial sugar content of 29.1 ° Bx. The fermentation rate until the fourth day was faster than that of GBY1 (based on the amount of alcohol produced), and the fermentation of fermivin was steadily continued until the last 16% (see FIG. 6) of alcohol was produced.

Fermentation of bokbunja during fermentation time Number of days of fermentation Alcohol(%) Brix Acid (g / L) pH L A B 0 0 30.7 9.02 3.83 13.98 46.04 24.44 One 2.73 25.5 11.11 3.69 6.12 34.73 10.55 2 8.76 18.6 12.34 3.68 7.06 37.03 12.18 3 12.01 15 12.31 3.78 7.01 36.89 12.08 4 13.94 12.9 12.86 3.79 7.1 36.91 12.24 5 14.87 11.9 13.13 3.78 6.85 36.51 11.81 6 15.3 11.6 13.22 3.77 6.76 36.12 11.66 7 15.33 11.5 13.18 3.8 6.54 35.78 11.28 8 15.29 11.5 13.09 3.84 6.57 35.89 11.32 9 15.24 11.4 12.67 3.85 6.58 35.84 11.34 Juice 15.2 11.4 12.54 3.89 6.92 36.47 11.94 percolation 14.94 11.2 12.48 3.88 6.61 35.64 11.39

Fermentation of Bacillus subtilis during the fermentation time of commercial strain ( S. cerevisiae Fermivin) Number of days of fermentation Alcohol(%) Brix Acid (g / L) pH L A B 0 0 29.1 8.67 3.58 14.83 46.7 25.56 One 0.52 28 9.57 3.54 12.54 44.06 21.62 2 4.33 23.3 11.64 3.45 11.99 43.34 20.67 3 10.7 16.4 11.84 3.52 11.67 43.17 20.12 4 13.58 13.3 12.16 3.55 11.67 43.16 20.11 5 15.17 11.6 11.85 3.63 11.4 42.81 19.65 6 15.71 10.9 11.66 3.61 12.39 44.15 21.36 7 16.07 10.9 12.04 3.61 11.4 42.88 19.65 8 16.12 10.8 11.94 3.62 11.28 42.77 19.45 9 16.1 10.8 11.96 3.65 11.19 42.64 19.29 Juice 16.09 10.8 12.29 3.65 11.18 42.72 19.28 percolation 16.04 10.8 11.98 3.66 10.94 42.3 18.86

GBY1, a strain isolated in the present invention, was found to have a fermenting ability similar to that of the commercial strain Fermivin, suggesting that GBY1 could be used for the macromolecular fermentation.

<Example 2> ODY alcohol fermentation using GBY1

20% sugar of raw material was added to the oat fruit, 600 ppm of enzyme viscozyme was added, 10 ⁹ CFU / mL of fruit / 1 kg of fruit was added, and the quality change was confirmed at 25 캜 fermentation at intervals of 1 day. When 20% of the fruits were fed, the sugar content was 33.1 ° Bx. When fermented at 25 ℃, the sugar content decreased to 29.8 ° Bx and the alcohol content was 1.49% on the first day. The sugar content was steadily decreased, and alcohol was produced and about 16.22% of alcohol was produced on the fifth day. When fermentation was completed, the final alcohol content was 17.88% and the sugar content was decreased to 11.4 ° Bx. When the same fermentation was carried out with the commercial strain Fermivin, the initial sugar content started from 30.2 ° Bx and the final alcohol content was 15.18% and the sugar content decreased to 11.1 ° Bx. The results are shown in Tables 3 and 2-2 below.

Audi fermentation over time of fermentation Number of days of fermentation Alcohol(%) Brix Acid (g / L) pH L A B 0 0 33.1 2.69 5.06 12.75 41.88 20.88 One 1.49 29.8 4.29 4.7 8.86 38.51 15.27 2 8.89 21.5 6.52 4.23 14.72 46.32 25.38 3 12.67 17.4 6.78 4.29 15.34 46.94 26.36 4 14.85 15.3 6.77 4.41 16.18 47.66 27.47 5 16.22 13.7 6.63 4.42 16.63 48.14 28.19 6 17.24 12.8 6.73 4.42 16.12 47.46 27.35 7 17.78 12.2 6.66 4.47 16.44 47.66 27.75 8 18.07 11.9 6.54 4.52 17.35 48.31 28.91 9 18.17 11.7 6.25 4.55 19.76 49.97 31.85 Juice 18.09 11.7 6.11 4.56 20.91 50.47 32.92 percolation 17.88 11.4 5.98 4.57 22.12 51.04 34.14

The fermentation time of commercial strains ( S. cerevisiae Fermivin) Number of days of fermentation Alcohol(%) Brix Acid (g / L) pH L A B 0 0 30.2 2.05 5.16 11.29 40.08 18.84 One 0.5 28.1 2.55 5.02 9.83 39 16.82 2 0.65 27.5 5.3 4.31 13.59 44.1 23.35 3 2.17 25.6 6.68 4.18 15.11 45.9 25.87 4 8.39 18.9 7.56 4.17 17.92 48.73 29.73 5 11.7 15.5 7.67 4.22 19.31 49.83 31.06 6 13.57 13.6 7.65 4.23 19.56 49.99 31.32 7 14.67 12.4 7.68 4.25 20.07 50.27 31.83 8 15.24 11.7 7.64 4.26 20.85 50.74 32.39 9 15.54 11.4 7.48 4.28 21.72 51.24 33.05 11 15.74 11.3 7.49 4.28 22.21 51.19 32.94 Juice 15.64 11.2 7.19 4.3 23.13 51.6 33.31 percolation 15.18 11.1 7.01 4.28 25.07 52.12 33.91

GBY1, a strain isolated in the present invention, was found to produce higher alcohol for a short time because it was more effective in fermentation than the commercial strain Fermivin.

<Example 3> Blueberry alcohol fermentation using GBY1

16% sugar of raw material was added to blueberry fruit, 600 ppm of enzyme viscozyme was added, 10 ⁹ CFU / mL of fruit / 1 kg of fruit was added, and the quality change was checked at 25 ° C intervals at intervals of 1 to 2 days. -2.

When 16% of the fruits were fed, the sugar content was 26.7 ° Bx. When fermented at 25 ℃, the sugar content decreased to 21.1 ° Bx and the alcohol content was 4.90% on the first day. The sugar content was steadily decreased, and alcohol was produced and about 12.56% of alcohol was produced on the sixth day. When fermentation was completed, the final alcohol was 15.45% and the sugar content was decreased to 10.1Bx. When fermented with the commercial strain Fermivin, the initial sugar content started at 36.5 ° Bx and the final alcohol content was 16.22%, and the sugar content decreased to 11.1 ° Bx. Blueberry fermentation using Fermivin was fermented for about 40 days. The fermentation rate of S. cerevisiae GBY1 was about twice as fast as that of GBY1 fermentation on the 6th day, similar to the alcohol production of 12.43% on fermentation on 14th day of fermentation .

Blueberry fermentation performance over time of fermentation Number of days of fermentation Alcohol(%) Brix Acid (g / L) pH L A B 0 0 26.7 9.02 3.06 54.27 60.52 2.43 2 4.9 21.1 11.74 2.86 50.11 63.51 6.43 4 9.65 16.3 12.65 2.96 50.56 62.47 6.06 6 12.56 13.1 13.32 2.97 50.52 62.07 6.12 8 14.23 11.3 13.36 3 49.9 61.89 5.71 9 14.73 10.8 13.44 3.04 50.82 60.92 5.24 12 15.32 10.2 13.24 3.08 49.69 60.78 5.13 14 15.38 10.1 13.22 3.11 53.53 57.33 3.7 Juice 15.45 10.1 13.07 3.1 52.95 57.47 4.16

Blueberry fermentation over time of fermentation of commercial strain ( S. cerevisiae Fermivin) Number of days of fermentation Alcohol(%) Brix Acid (g / L) pH L A B 0 0 36.5 6.55 3.21 56.6 55.91 3.33 One 0.6 31.3 7.65 3.12 45.13 63.03 9.78 2 2.44 27.6 9.4 3.03 44.05 64.21 10.84 3 4.31 25.1 9.84 3.09 43.93 63.94 10.83 4 5.48 23.3 10.07 3.14 42.62 64.37 12.12 5 6.3 21.7 10.18 3.15 46.23 62.21 8.26 6 7.35 20.6 10.36 3.16 46.76 61.47 7.7 7 8.25 20.1 10.7 3.1 42.04 63.96 11.26 8 9 19.3 10.91 3.16 42.11 63.8 12.08 9 9.63 18.7 11.01 3.15 45.66 61.68 8.28 10 10.3 17.8 11.23 3.11 43.4 63.16 10.87 11 10.91 17.1 11.25 3.13 42.92 63.16 10.87 12 11.34 16.4 11.29 3.19 45.62 61.32 8.61 13 11.45 15.7 11.05 3.19 46.38 60.76 8.54 14 12.43 15.5 11.32 3.19 42.44 62.67 11.06 15 12.9 15.2 11.46 3.22 43.34 62.21 11.11 16 13.27 14.8 11.58 3.23 43.11 62.15 10.63 17 13.7 14.4 11.57 3.22 43.18 61.95 10.59 18 14.05 14 11.71 3.25 43.29 61.81 10.43 19 14.21 13.6 11.64 3.27 46.62 59.71 8.11 21 14.9 13.1 11.84 3.29 44.34 60.86 10.17 23 15.36 12.5 11.75 3.32 44.83 60.36 9.52 25 15.69 12.1 11.79 3.3 46 59.42 9.27 28 16.04 11.6 11.74 3.3 46.69 58.52 9.11 30 16.22 11.5 11.64 3.28 46.9 58.37 7.91 32 16.29 11.5 11.75 3.29 47.86 57.52 8.16 35 16.39 11.3 11.72 3.33 49.63 55.9 8.13 37 16.38 11.2 11.81 3.31 49.75 55.51 8.53 Juice 16.3 11.2 11.71 3.32 50.4 54.99 8.51 percolation 16.22 11.1 11.67 3.2 52.71 53.25 8.22

GBY1, a strain isolated in the present invention, was found to be excellent for blueberry fermentation.

The strain of the present invention ( Saccharomyces cerevisiae GBY1, KCCM11417P) showed excellent results in ethanol fermentation of berry, vermiculite, and blueberry. In addition, it has been found that the fermentation rate is faster and the yield per unit is higher than that of commercial strains, which is highly likely to be used industrially.

Korea Microorganism Conservation Center (overseas) KCCM11417P 20130522

Claims (5)

Wherein the yeast is Saccharomyces cerevisiae GBY1, Korean Microorganism Conservation Center Accession No: KCCM11417P) isolated from Gochang San and the yeast is fermented with blueberry. delete Fermenting berries by thawing and crushing berries and adding saccharides to adjust sugar content and inoculating yeast ( Saccharomyces cerevisiae GBY1, Korean Microorganism Conservation Center Accession No: KCCM11417P, 2013. 5.22 deposit) of claim 1 and fermenting Wherein the berries are blue berries. The method of claim 1, wherein the berries are blue berries. The method according to claim 3, wherein the saccharified berries are subjected to fermentation by feeding sucrose at 20% to the fruit and by inoculating 1 × 10 ⁹ CFU / mL of yeast per 1 kg of fruit. . delete

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