KR102088696B1 - Saccharomyces cerevisae BA34 strain for manufacturing the wine using various berries and not producing biogenic amine and uses thereof - Google Patents

Abstract

The present invention relates to a non-producing Saccharomyces cerevisiae BA34 strain and its use for the production of berry wines, in the present invention, Saccharomyces cerevisiae isolated from berry fruits cerevisiae ) It was confirmed that the BA34 strain is resistant to alcohol, sugar, and sulfurous acid, has alcohol-producing ability, and β-glucosidase secretion ability, and does not produce biogenic amines and ureases. Therefore, it is expected that the Saccharomyces cerevisiae BA34 strain of the present invention can provide a safe wine free of biogenic amines, and thus can be usefully used industrially.

Description

Biogenic amine for the production of berry wines Saccharomyces cerevisiae BA34 strain and its use {Saccharomyces cerevisae BA34 strain for manufacturing the wine using various berries and not producing biogenic amine and uses thereof}

The present invention relates to a non-producing Saccharomyces cerevisiae BA34 strain and its use for the production of berry wines.

In Korea, fermentation technology and aging technology used in the manufacture of food have been developed since ancient times, and accordingly, traditional alcoholic beverages using fermentation technology have been continuously developed. Traditional liquors are largely divided into traditional distilled liquor and traditional fermented liquor according to the manufacturing method. In particular, traditional fermented liquor has a unique taste and aroma according to the added ingredients, and each has unique functionality.

There fermentation of traditionalism include saccharide as MY access (Saccharomyces) genus Aspergillus (Aspergillus) genus Bacillus (Bacillus) genus Lactobacillus bacteria (Lactobacillus) in such strains have been used, in recent years, with fermented material such fermentation strain Health functional fermented liquor is being produced which emphasizes the efficacy and characteristics of byproducts produced during the fermentation process.

Gamma-amino butyric acid (GABA), one of the products produced by fermentation microorganisms during fermentation, is a non-proteinaceous amino acid that is distributed in nature and mainly inhibits the central nervous system, such as the brain or spinal cord. It is known as an inhibitory neurotransmitter. Due to various pharmacological activities such as activating the blood flow of the brain and increasing the oxygen supply to the brain to promote the metabolic function of brain cells, reducing triglycerides in the blood and improving liver function, headaches and ringing caused by cerebral arterial idleness , It has been applied to therapeutic agents such as decreased motivation. In addition, it has been found to be effective in the prevention and improvement of adult diseases such as hypertension, increasing interest as a functional food material, and recently, beverages containing a large amount of gamma-aminobutyric acid have been released.

On the other hand, biogenic amine (biogenic amine, BA) is responsible for physiological functions such as growth control, inflammation control, and neurotransmission in the human body, but when the biogenic amine exceeding the decomposition limit of the human body is consumed through food, there are harmful symptoms. appear. Therefore, in fermentation, it is very important to select a fermentation strain that does not produce biogenic amine during the fermentation process.

A variety of microorganisms are involved in the fermentation of berry-like alcohol, such as wine, and are made through various actions between microorganisms while repeating complex chemical steps. Many overseas countries use Saccharomyces cerevisiae as a starter strain for the production of wine to improve the quality of wine, and for the selection of yeast that has excellent alcohol fermentation ability and helps to improve the quality of wine production. There is a lot of effort.

On the other hand, Korean Registered Patent No.1599271 discloses 'Saccharomyces cerevisiae BA42 strain that produces alcohol separated from berry fruits and does not produce biogenic amines and uses thereof', and Korean Registered Patent No. 1599270 In 'Saccharomyces cerevisiae BA33 strain that produces alcohol separated from Audi and does not produce biogenic amine and its use' is disclosed, as in the present invention, 'Biogenic amine for berry wine production No non-producing Saccharomyces cerevisiae BA34 strain and uses thereof have been found.

The present invention has been derived by the above-mentioned demands, in the present invention, it is resistant to alcohol, sugar, and sulfurous acid from berry fruits, has alcohol-producing ability, and β-glucosidase secretion ability, and biogenic amine. And Saccharomyces cerevisiae that does not produce urease (Saccharomyces cerevisiae) BA34 strain was isolated.

Accordingly, the present invention was completed by confirming that the Saccharomyces cerevisiae BA34 strain of the present invention can be variously used in fermented food related industries such as starter strains for wine production.

In order to solve the above problems, the present invention is resistant to alcohol, sugar and sulfurous acid, has alcohol-producing ability and β-glucosidase secretion ability, and does not produce biogenic amines and ureases. Saccharomyces cerevisae BA34 strain (KCCM12243P) is provided.

In addition, the present invention provides a composition for producing alcohol containing the strain, a culture solution thereof, a concentrate solution of the culture solution, or a dried product thereof as an active ingredient.

In addition, the present invention provides a starter composition for preparing fermented food comprising the strain, a culture solution thereof, a concentrate solution of the culture solution, or a dried product thereof as an active ingredient.

In the present invention, the Saccharomyces cerevisiae BA34 strain isolated from berry fruits is resistant to alcohol, sugars and sulfites, has alcohol production capacity and β-glucosidase secretion ability, It was confirmed that no biogenic amine and urease were produced. Therefore, it is expected that the Saccharomyces cerevisiae BA34 strain of the present invention can provide a safe wine free of biogenic amines, and thus can be usefully used industrially.

Figure 1 shows a schematic diagram of the 18S rRNA gene base of Saccharomyces cerevisae BA34 strain (KCCM12243P) isolated from the present invention.
Figure 2 is a result of analyzing the resistance to different concentrations of ethanol, sugar and sulfurous acid of the Saccharomyces cerevisae BA34 strain (KCCM12243P) isolated from the present invention. After incubation at 30 ° C, 200 rpm, and 35 hr in YPD medium containing different concentrations of ethanol, sugar, and sulfurous acid, resistance of Saccharomyces cerevisiae was exhibited by dry cell weight (g / L). (A) Saccharomyces cerevisiae resistance to different ethanol concentrations (0%, 8%, 14%, and 20%), (B) Different glucose concentrations (3%, 30%, 40%, and 50%) Saccharomyces cerevisiae resistance to), (C) Saccharomyces cerevisiae resistance to different sulfite concentrations (0 ppm, 100 ppm, 200 ppm, and 300 ppm). This is the average value of the experiments performed with 3 replicates.

In order to achieve the object of the present invention, the present invention is resistant to alcohol, sugar and sulfurous acid, has alcohol-producing ability and β-glucosidase secretion ability, and does not produce biogenic amines and ureases. Provided is the Saccharomyces cerevisae BA34 strain (KCCM12243P).

Saccharomyces cerevisiae BA34 strain according to the present invention produces alcohol from strains separated from various berry fruits and extracts such as Audi, Murru, Bokbunja, Blueberry grown and harvested in Sunchang-gun, Jeollabuk-do, Audi wine, Bokbunja When producing wine and blueberry wine, it does not produce biogenic amine, and is resistant to alcohol, sugar, and sulfurous acid, so it is selected as the most suitable strain as a fermentation strain for berry wine production.

As a result of sequencing of the 18S rRNA region of the selected BA34 strain of the present invention, Saccharomyces cerevisiae cerevisiae ).

The strain Saccharomyces cerevisiae BA34 of the present invention was deposited with the Korea Microbial Conservation Center on March 28, 2018 (Accession No .: KCCM12243P).

In a strain according to an embodiment of the present invention, the biogenic amine is histamine, putrescine, cadaverine, spermidine, spermine, tryptamine , Hisiseamine, 2-phenylethylamine, tyramine, serotonin, L-norepinephrine or dopamine, preferably histamine , Tyramine, putrescine or cadaverine, but is not limited thereto.

In addition, the present invention provides a composition for producing alcohol containing the strain or a culture medium thereof as an active ingredient.

In the composition according to an embodiment of the present invention, the alcohol may be alcohol fermented grains or fruits, most preferably wine fermented berries, but is not limited thereto.

The method for culturing the strain of the present invention may be cultured according to a method commonly used in the art, and is not limited to a special method.

When using the concentrate obtained from the step of culturing the strain of the present invention or the culture of the strain or the concentrate of the culture medium of the strain as an additive, the culture medium of the strain or the strain or the culture medium of the strain is added as it is, or Other additives may be used together, and may be suitably used according to conventional methods. The mixing amount of the active ingredients may be appropriately determined according to the purpose of use.

In addition, the present invention provides a starter composition for preparing fermented food comprising the strain, a culture solution thereof, a concentrate solution of the culture solution, or a dried product thereof as an active ingredient.

In the present invention, a starter for preparing fermented food means a preparation or composition containing microorganisms involved in fermentation for the production of fermented food. It is used to provide microorganisms that can grow as a dominant species or microorganisms that can grow in fermented foods by adding them during the production of fermented foods. When the food is prepared using the food fermentation starter, the quality of the food is constantly controlled by the microorganisms contained in the food fermentation starter, or a specific purpose, for example, does not cause or reduce odor in food. You can achieve your purpose. In the present invention, by using the Saccharomyces cerevisiae BA34 strain or a culture medium thereof as a starter strain, alcohol and β-glucosidase (β-glucosidase) are produced, and a fermented food that does not produce biogenic amine and urease Can be produced.

In the starter composition for preparing fermented food according to one embodiment of the present invention, the fermented food may be fermented wine fermented with grains or fruits, wine fermented with berry, kimchi, jang or fermented milk, preferably wine fermented with berry It may be, but is not limited thereto.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

Materials and methods

Yeast separation and culture conditions

In order to separate various yeasts, various berry fruits and extracts such as audi, fruit, bokbunja, and blueberries grown and harvested in Sunchang-gun, Jeollabuk-do were purchased and used. After homogenizing 10 g of berry fruits and extracts, diluting them in decimal with 100 ml of sterile physiological saline, YPD (100% yeast extract 1%, peptone 2%, glucose) containing 100 μg / ml chloramphenicol to suppress bacterial growth. 3%) Plated on a solid medium and incubated at 30 ° C for 2 days. To separate pure yeast colonies, single colonies were isolated and incubated for 2 days at 30 ° C. in YPD liquid medium. Among the yeast strains, biogenic amine non-producing yeast suitable for berry wine fermentation was selected, mixed with YPD liquid medium containing 20% glycerol and stored at 4 ° C. Cell activation was performed by incubating for 30 days at 30 ° C in a YPD liquid medium.

Separation of alcoholic fermentable yeast

The gas generating ability was confirmed through the Durham test. For cultivation of the selected strains, yeast was cultured by stirring in YPD liquid medium at 30 ° C. for 24 hours. 2% of the culture was inoculated into a Duram tube containing 5 ml of YPD liquid medium containing 20% glucose. After that, all the Duram test tubes were incubated for 48 hours at 30 ° C without agitation, and after 48 hours, the gas was first checked by checking whether gas was generated, and the primary selection yeasts were inoculated in YPD containing 20% glucose, and the culture solution was cultured after 48 hours of incubation After centrifugation, the culture supernatant was taken and filtered using a 0.45 μm syringe filter, and then alcohol generation ability was analyzed using HPLC according to the conditions according to Table 1.

HPLC analysis conditions used to detect biogenic amines, glucose and ethanol HPLC analysis conditions used for biogenic amine detection Instrument Agilent 1200 series
(Agilent Technologies, Santa Clara, CA, USA) Column CapcellPak C18 Column Detector DAD detector (254 nm) Mobile phase A: 0.1% formic acid in H ₂ O B: 0.1% formic acid in ACN Gradient condition A: B = 45:55, 0 ~ 10 min A: B = 35:65, 10 ~ 15 min A: B = 20:80, 15 ~ 20 min A: B = 10:90, 20 ~ 30 min A: B = 10:90, 40 min over Flow rate 1.0 ml / min Temperature 40 ℃ Injection volume 20 μL HPLC analysis conditions used for glucose and ethanol detection Instrument Agilent 1200 series
(Agilent Technologies, Santa Clara, CA, USA) Column Aminex HPX 87H column, 300 Х 7.8 mm
(Bio-Rad, Hercules, CA, USA) Detector RID detector Eluant 5 mM Sulfuric acid in H ₂ O Flow rate 0.6 ml / min Temperature 65 ℃ Injection volume 10 μL

Biogenic Amine  Check creation

Investigation as to whether or not to generate biogenic amines of the selective isolates, after inoculating each isolate into the YPD liquid medium, and pre-incubating for 24 hours at 150 rpm in a shaking incubator at 30 ° C, 1 ml of the culture medium is a precursor amino acid of tyramine, histamine, putresin, and cadaverine. Tyrosine, histidine, arginine, and lysine were inoculated into 9 ml of a YPD liquid medium containing 0.1% and incubated for 48 hours in a 30 ° C shaking incubator 150 rpm. The culture medium was centrifuged at 13,000 rpm for 30 minutes to remove the cells, and the wine produced using the culture supernatant and berries from which the cells were removed was used as a sample for biogenic amine analysis. 0.5 ml of sample solution and standard solution were respectively taken, and then 0.25 ml 1,7-diaminoheptane (Sigma-aldrich) and 0.25 ml saturated Na ₂ CO ₃ The solution (Sigma-aldrich), 1% acetone (Sigma-aldrich), and 0.4 ml monosyl chloride (Sigma-aldrich) were mixed and then derivatized at 45 ° C. for 1 hour. After adding 0.25 ml of 10% proline (Sigma-aldrich) to the derivatized sample, remove the residual amount of monosyl chloride, add 2.5 ml of ethyl ether (Samchun, Seoul, Korea), shake for 3 minutes, and then separate the supernatant. It was taken, evaporated, and the remaining residue was dissolved in 0.5 ml acetonitrile, filtered through a 0.45 μm syringe filter (Sartorius), and used for analysis. Instrumental analysis conditions for biogenic amine analysis are shown in Table 1.

Urease Confirmation of creation

The generation of the urease, a harmful enzyme, was confirmed using a urea rapid test kit (MB cell, Korea). The urea rapid test kit was inoculated with the selected strain, and petroleum was added to it, and petroleum was incubated at 37 ° C. for 24 hours in a state where oxygen was blocked, and color change was observed at intervals of 4 hours to confirm whether urease was produced.

β- Glucosidase  Enzyme activity

To analyze β-glucosidase enzyme activity, lactobacilli MRS agar plates prepared by adding 1.0% esculin and 0.5% ferric ammonium citrate were inoculated with isolates. After incubation at 37 ° C for 24 hours, β-glucosidase enzyme activity was investigated depending on the presence or absence of a black transparent ring formed around the colony.

Identification of yeast and creation of phylogenetic tree

Finally, yeast BA34 suitable for fermentation of berry wine was selected from the selected strains, and DNAs were extracted to identify the selected strains, and the 18S rRNA gene was analyzed and requested by Cosmogene tech. The nucleotide sequence was obtained through the ExTaxone-e server ( http://www.eztaxon.org ), and the nucleotide sequence of the standard strain was obtained, and the MEGA 6.0 program was used to compare the nucleotide sequences to create a systematic diagram. Neighbor-joining algorithm was used for the tree diagram, and the robustness of the tree diagram created by bootstrapping at 1,000 iterations was confirmed.

Alcohol resistant, Sugar resistance , Sulfite resistance investigation

To investigate the alcohol resistance of Saccharomyces cerevisiae BA34, 0, 8, 14, and 20% anhydrous ethanol were added immediately after inoculating yeast into the YPD liquid medium. Dry cell mass was investigated by incubation at 30 ° C for 72 hours. Glucose tolerance was investigated by incubating yeast in a YPD liquid medium containing 3, 30, 40, and 50% glucose at 30 ° C. for 48 hours and measuring the amount of dry cells. The sulfurous acid resistance was measured by incubating for 48 hours at 30 ° C in a YPD liquid medium to which potassium metabisulfite of 100, 200 and 300 ppm was added to measure the amount of dry cells.

Berries  Used wine  Produce

Berry was inoculated with yeast selected from YPD liquid medium to produce wine, and cultured at 150 rpm in a 30 ° C shaking incubator for 24 hours, and the culture was centrifuged at 13,000 rpm for 20 minutes. The separated cells were washed three times in sterile distilled water and used as a starter for berry wine production. The berry fermentation broth was used by sterilizing the sugar content to 12 ° Brix by adding white sugar to the stock solution filtered through filter paper by pulverizing Audi, bokbunja, and blueberry fruits, respectively. Prior to this, 2% (v / v) of the cultured yeast was inoculated, and a fermentation broth cultured with shaking at 150 rpm in a 30 ° C shaking incubator for 48 hours was used as a mother. The production of berry wine is adjusted to 22 ° Brix by adding white sugar, and then inoculated with 2% (v / v) of the mother in sterile fruit juice, fermented for 5 days at 25 ~ 30 ℃, and aged through a 7-day aging period. Did.

Example  One. Berries  Separation of yeast from fruit

In the present invention, yeast isolates were selected from berry fruits and extracts for separation of yeast strains, and the isolates were stored at -20 ° C for the next study and used in the next study.

Example  2. Gas and alcohol Generating ability  Selection of yeast to have

The experiment was conducted on the selective isolates, and gas formation in the Duram test tube was observed after incubation for 48 hours in a 30 ° C stationary incubator. Gas formation was investigated using all culture supernatant, and strains forming gas were first selected. However, according to the report, since most yeasts play a role of producing alcohol without an identifiable gas-forming ability, additionally, HPLC analysis was conducted to determine whether alcohols were produced, and as a result, 7 yeasts having an alcohol generating ability of 5% or more were finally obtained. Tea was selected (Table 2).

Ethanol content of strains selected from YPD liquid medium containing 24% glucose Strain Ethanol content (%) Commercial yeast (fruit wine) 10.28 ± 0.01 Commercial yeast (Gokju) 10.52 ± 0.05 BBA 03 8.78 ± 0.01 BBA 19 5.87 ± 0.01 BBA 23 9.65 ± 0.03 BA34 12.63 ± 0.05 BBA 46 8.63 ± 0.03 BBA 88 5.06 ± 0.01 BBA 91 6.06 ± 0.03

Example  3. Biogenic Amine Non-generation  Yeast screening

For the yeast that was selected as the first step, it was investigated whether or not biogenic amine was produced in the liquid medium containing the biogenic amine precursor, and secondly, after making wine using berries, the contents of histamine, tyramine, putrescine and cadaverine were investigated. Did. As a control, a juice solution of berries was used in consideration of the possibility of detection of biogenic amine present in the berries of itself, and berry wine was prepared to confirm whether or not to produce bionic amines by selective yeast. As a result of Vasantha Rupasinghe and Clegg, it was confirmed that histamine was also present in the fruit itself, and the results are shown in Table 3.Biogenic amine was detected by some strains as well as in the berry juice itself, but mostly quantitative In the case of BA34, since all four biogenic amines were not detected in wines produced using berries, BA34 was selected as a suitable strain for wine production using berries.

Results were measured 3 times and indicated as standard deviation

^a YPD liquid medium containing only the juice itself or the precursor as ^a control

^b ND-not detected

^c NQ-not quantified

Example  4. Berries Wine  Alcohol content analysis

The results of analyzing the alcohol content by preparing berry wines using seven strains of BBA03, BBA19, BBA23, BA34, BBA46, BBA88, and BBA91 identified as Saccharomyces cerevisiae are shown in Table 4. Ethanol and blueberries, ethanol was not detected in the control group without inoculation of the strain into the bokbunja stock solution, and in the case of Audi wine, the commercial yeast (grain) inoculated with the control was the highest with 18.82%, and among the selected strains, the BBA23 strain The wine produced by inoculation was the highest with an alcohol content of 17.01%. In blueberry wine, 6 strains except BBA03 selected were higher than 2 commercial yeasts inoculated with control. In the case of double-molecule wine, BA34 was the highest among the selected strains including the control at 18.68%. In the case of blueberry wine, unlike Audi and bokbunja, both showed an alcohol content of 5% or more, but due to the high acidity of the raw material blueberry, the yeast strain itself has difficulty in growing, thereby reducing sugar consumption of the selected yeast. The production of alcohol by is thought to be poor.

Ethanol content in wines made from various berries using selected strains Strain Ethanol content (%) Audi Blueberries Bokbunja Control N.D. N.D. N.D. Commercial yeast (fruit wine) 18.33 ± 0.05 5.68 ± 0.03 17.54 ± 0.02 Commercial yeast (Gokju) 18.82 ± 0.03 5.39 ± 0.01 15.55 ± 0.01 BBA 03 15.55 ± 0.01 4.79 ± 0.01 15.01 ± 0.02 BBA 19 16.37 ± 0.01 6.96 ± 0.05 13.82 ± 0.05 BBA 23 17.01 ± 0.03 7.43 ± 0.05 17.49 ± 0.01 BA 34 16.59 ± 0.01 7.94 ± 0.05 18.68 ± 0.01 BBA 46 16.79 ± 0.01 8.62 ± 0.01 15.27 ± 0.02 BBA 88 15.25 ± 0.01 7.08 ± 0.01 13.03 ± 0.05 BBA 91 15.48 ± 0.05 7.21 ± 0.03 13.72 ± 0.05

Example  5. Urease ( Urease ) Confirmation of creation

As a mechanism of cell damage, cell infection is caused by ammonia produced by urease when Helicobacter pylori infection, and phobias of epithelial cells are caused by Vac A. Inflammatory substances secreted from neutrophils and activity Cells are also damaged by oxygenates. Damage to epithelial cells causes atrophic changes in cells, cell proliferation increases as a compensation mechanism for cell damage, and cells that actively proliferate have a greater chance of damaging genes by carcinogens. Therefore, an experiment was conducted to check whether urease was generated, and it was confirmed that urease was not generated in the BBA19, BA34, BBA46, and BBA91 strains (Table 5).

Comparison of urease activity against selected yeast strains Strain Urease activity BBA 03 + BBA 19 - BBA 23 + BA34 - BBA 46 - BBA 88 + BBA 91 -

Example  6. β- Glucosidase  Enzyme activity

β-glucosidase (β-glucosidase) is known to catalyze the formation of non-glycosides and hydrolysis of isoflavone glycosides. As a microorganism that produces β-glucosidase, Aspergillus niger , Trichoderma reesei , Fusarium oxysporum ( Fusarium) molds such as oxysporum ) and Saccharomyces cerevisiae ), Schizosaccharoyces pombe Streptomyces reticuli , Bacillus subtilis , Bacillus polymyxa , Bacillus circulatory ( Bacillu) circulans ), Agrobacterium tumefaciens ), Cellulomonas sp.CS1-1 and strains belonging to various lineages such as Pyrococcus furiosus , an ultra-high temperature bacterium, are known, and for the β-glucosidase they produce Enzymatic properties and genes have also been identified. As a result of confirming the activity of β-glucosidase having bioavailability in the bioconversion process to convert into non-glycoside form for absorption by the soybean isoflavone glycosides, genistin and didazine, which are mainly present in traditional fermented foods, β- for BA34 strain It was confirmed that the activity of glucosidase was excellent (Table 6).

Comparison of β-glucosidase activity against selected yeast strains Strain β-glucosidase activity BBA 03 ++ BBA 19 - BBA 23 + BA34 +++ BBA 46 + BBA 88 ++ BBA 91 +

Example  7. Identification of yeast and creation of phylogenetic tree

For identification of the selected strain BA34, the gene was amplified using ITS1 and ITS4 primers to analyze the 18S rRNA gene sequence. As a result of BLAST in GenBank, the analyzed result was identified as S. cerevisiae , and the nucleotide sequence is shown in SEQ ID NO: 1. In the SeqMatch program, nucleotide sequences were used to perform cross-comparison with highly homologous standard strains. Phylogenetic trees were prepared based on the 18S rRNA gene base sequence (FIG. 1), and finally designated as Saccharomyces cerevisiae BA34. The final selected strain, BA34, is Saccharomyces cerevisiae at the Korean Culture Center of Microorganisms (KCCM). Deposited with KCCM12243P.

Example  8. Alcohol resistance, sugar resistance and sulfurous acid resistance of selected strains

Saccharomyces cerevisiae strains are commonly used because of the production of aromatic scent components and increased sulfite and alcohol resistance. Therefore, alcohol, sugar, and sulfurous acid resistance are essential elements as yeast for wine production due to the addition of high concentrations of glucose, alcohol, and sulfurous acid in the production of grape wine. Therefore, the resistance of alcohol, sugar, and sulfurous acid was examined for the resistance of selected strains to confirm the possibility as yeast for wine production using berries.

To investigate the alcohol resistance of BA34, 0, 8, 14, and 20% anhydrous ethanol were added to the YPD liquid medium, which is a general yeast culture medium, and the dry cell mass was examined to confirm resistance. The amount of dried cells was as shown in Fig. 2A, and in the experimental group with 8% added, it decreased to 2.78 g / L compared to the control (6.63 g / L) and hardly grown in the high concentration added group. This confirmed results similar to those of previous studies that the final biomass concentration was low when the initial glucose and alcohol concentrations were high. However, compared to other literature on alcohol resistance, it did not show a significant difference from other research results because it did not grow well at a concentration of 8% or more.

The sugar resistance was as shown in FIG. 2B, and significantly increased to 10.13 g / L in the experimental group with 30% glucose added, compared to the cell mass (6.04 g / L) of YPD, a general yeast culture medium. When more than 4% of sugars suggested by Calado et al. (2003) and Kim were added, the results showed that the cell mass and alcohol production capacity were low. On the other hand, 40% and 50% glucose was added to the experimental group decreased to 9.16 g / ℓ and 8.99 g / ℓ, but it was confirmed that even in the experimental group to which high concentration of glucose was added, it was not affected much by growth. However, as the addition rate of glucose increased, the cell growth rate decreased as a result of the difficulty in circulation of water in the culture medium due to the decrease in biomass production and the high degree of sugar addition, the dry cell mass was also decreased.

Sulfuric acid is a substance that is added for the preservation of food, beverages, and drugs, and is used because of its anti-oxidation, anti-bacterial, and browning-resistant effects. Therefore, it is used to prevent oxidation and selectively inhibit harmful microorganisms in wine production. Therefore, yeast needs to have sulfite resistance for wine production. According to the current domestic food standard, the results of investigating resistance by adding sulfuric acid of 100, 200, 300 ppm in accordance with the allowable specification of 350 mg / ℓ are as shown in FIG. 2C. In comparison, although the tendency of slight decrease was 4.64 g / ℓ, 4.11 g / ℓ, and 4.52 g / ℓ, it was confirmed that it can be used as yeast for berry wine production because it is not greatly affected.

Korea Microbial Conservation Center (Overseas) KCCM12243P 20180328

<110> Microbial Institute for Fermentation Industry <120> Saccharomyces cerevisae BA34 strain for manufacturing the wine          using various berries and not producing biogenic amine and uses          thereof <130> PN18147 <160> 1 <170> KopatentIn 2.0 <210> 1 <211> 813 <212> DNA <213> Saccharomyces cerevisae <400> 1 ggaaggatca ttaaagaaat ttaataattt tgaaaatgga ttttttttgt tttggcaaga 60 gcatgagagc ttttactggg caagaagaca agagatggag agtccagccg ggcctgcgct 120 taagtgcgcg gtcttgctag gcttgtaagt ttctttcttg ctattccaaa cggtgagaga 180 tttctgtgct tttgttatag gacaattaaa accgtttcaa tacaacacac tgtggagttt 240 tcatatcttt gcaacttttt ctttgggcat tcgagcaatc ggggcccaga ggtaacaaac 300 acaaacaatt ttatttattc attaaatttt tgtcaaaaac aagaattttc gtaactggaa 360 attttaaaat attaaaaact ttcaacaacg gatctcttgg ttctcgcatc gatgaagaac 420 gcagcgaaat gcgatacgta atgtgaattg cagaattccg tgaatcatcg aatctttgaa 480 cgcacattgc gccccttggt attccagggg gcatgcctgt ttgagcgtca tttccttctc 540 aaacattctg tttggtagtg agtgatactc tttggagtta acttgaaatt gctggccttt 600 tcattggatg tttttttttc caaagagagg tttctctgcg tgcttgaggt ataatgcaag 660 tacggtcgtt ttaggtttta ccaactgcgg ctaatctttt ttatactgag cgtattggaa 720 cgttatcgat aagaagagag cgtctaggcg aacaatgttc ttaaagtttg acctcaaatc 780 aggtaggagt acccgctgaa cttaagcata tca 813

Claims (5)

delete delete It is resistant to alcohol, sugar and sulfurous acid, has alcohol-producing ability and β-glucosidase secretion ability, histamine, tyramine, putrescine, and cadaverine. Saccharomyces cerevisae BA34 strain (KCCM12243P) that does not produce biogenic amines and ureases of KCCM12243P, a culture medium thereof, a sugar concentration of 40-50% containing the culture medium concentrate or a dried product thereof as an active ingredient Composition for producing alcohol in the. The composition of claim 3, wherein the alcohol is berry wine. delete

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