KR20230087738A - Novel Wickerhamomyces anomalus KJ-1 strain excellent in sugar tolerance, alcohol tolerance and fragrance production ability and uses thereof - Google Patents

Abstract

The present invention relates to a novel wickerhamomyces anomalus KJ-1 strain (accession number KACC 93373P) excellent in sugar tolerance, alcohol tolerance and fragrance production ability and uses thereof. When manufacturing alcoholic beverages using the Wickerhamyces anomalus KJ-1 strain (accession number KACC 93373P), the aroma components are enhanced without loss of aroma components, making it possible to manufacture the alcoholic beverages with excellent sensory properties, and the strain can grow without being killed under high-concentration sugar or alcohol conditions, and has antioxidant activity and antihypertensive activity.

Description

Novel Wickerhamomyces anomalus KJ-1 strain excellent in sugar tolerance, alcohol tolerance and fragrance production ability and uses thereof

The present invention relates to a novel Wickerhamomyces anomalus KJ-1 strain having excellent sugar tolerance, alcohol tolerance and fragrance production ability, and uses thereof.

Liquor is any beverage that contains more than 1% alcohol. Alcohol is naturally occurring and has developed into various forms of individuality according to regional, ethnic, climate, climate and cultural differences. Since there was no modern fermentation technology before the existence of molds and yeasts was discovered, fermented liquor was prepared by native molds and yeasts present in yeast. However, there was a problem in that it was difficult to maintain excellent and constant fermentation ability when using native yeasts. In order to solve this problem, fermentation was performed by isolating and selecting yeasts having excellent characteristics among native yeasts, thereby maintaining uniform fermentation ability and producing excellent fermented liquor.

In addition, alcohol fermentation of sugar (starch) is essential for the production of alcohol, and for this purpose, yeast suitable for the characteristics of each alcoholic beverage is used. Beer has cohesiveness for low-temperature fermentation and reuse, whiskey has high-temperature fermentation and alcohol production, and spirits and vodka have high alcohol production while minimizing components other than alcohol as fermentation characteristics. Whiskey, brandy, rum, gin, vodka, tequila, and soju are all alcoholic beverages made with carefully selected yeast, and research to find and industrialize the optimal yeast for each alcoholic beverage's characteristics is continuously being conducted.

On the other hand, in the manufacture of alcoholic beverages, raw materials are used to undergo an alcohol fermentation process. During the fermentation process, various by-products are produced depending on the type of raw materials, fermentation conditions, strains used, and the like. Depending on the type of these by-products, fermented products and alcoholic beverages using them have unique flavors. Bitter taste is alcohol, sour taste is lactic acid, succinic acid and trace amounts of acetic acid, sweet taste is sugars mainly composed of glucose and various amino acids, and fragrance is Higher alcohol acids and esters, and colors are expressed by flavin, deferichrome, and melanin.

Among them, an important factor influencing the quality of alcoholic beverages is fragrance, and most of the components that affect the fragrance of alcoholic beverages are determined by fermentation strains. Therefore, research is being conducted to discover fermented strains with excellent flavor for the production of alcoholic beverages with excellent flavor. However, in order to manufacture a variety of high-quality alcoholic beverages, it is necessary to secure various fermentation strains, and more improvements and development of new strains are required.

In addition, among the types of alcoholic beverages, distilled spirits are made by distillation to obtain alcohol with a higher alcohol content than fermented spirits, and include whiskey, brandy, rum, gin, vodka, tequila, and distilled soju. contain Distilled soju, one of Korea’s representative distilled spirits, is manufactured by mixing, fermenting, and distilling starch-containing raw materials and water, and the raw materials for distilled soju are grains such as rice, barley, and corn containing a lot of starch, or potatoes and sweet potatoes. documents, etc.

The vacuum distillation method is most commonly used for the production of such distilled spirits. The vacuum distillation method generally consists of a still, a single concentrator, and a single collection tank to carry out the distillation process, and in a single collection tank, the first, main, and downstream streams are produced. each are collected. In this existing device, even if the vinegar is removed, flavor components that adversely affect liquor quality remain in the distillation tube, inside the connecting pipe, and in the collection tank. In the structure with this structure, it is impossible to control the fractionation of the first and second streams, resulting in distillation products with poor main quality.

In addition, in the conventional cold-cold distillation, high-temperature steam is applied for a long time to increase the distillation ratio, resulting in a continuous increase in the temperature of the fermentation broth, resulting in the generation of heated odor, taste, and off-flavor, and a large amount of high-boiling components are collected, resulting in a distillate product of poor main quality. In order to offset this, an additional long-term aging process or an artificial aging process such as oak barrel aging is required, so it is highly likely that it will take a long time from distillation to production.

In addition, looking at the expression characteristics of important fragrance components in distilled spirits, most of the substances produced during the alcohol fermentation process in distilled spirits made of starch are ethyl alcohol or a small amount of amino acids and fatty acids in starch raw materials formed by the action of yeast It consists of fragrance ingredients. Fragrance components include scents that are sensually preferred and odors that cause unpleasant odors. In the case of preferred fragrance components, there are many low boiling point components. In the case of acetaldehyde, which is a component, it has a strong irritating odor, causes discomfort, and also causes a hangover. In addition, distilled spirits have a problem in that a large amount of volatile flavor components are lost and reduced during the distillation process.

Therefore, it is necessary to discover new fermentation strains with excellent productivity of fragrance components in order to produce alcoholic beverages that are not only excellent in consumer preference but also have good fragrance.

In addition, during alcohol fermentation, producing high-concentration alcohol in the fermentation tank is important not only to improve alcohol productivity, but also to improve the economics of the distillation step for recovering alcohol. In order to produce high-concentration alcohol, yeast must not be killed in the produced alcohol, and death by osmotic pressure should not occur in the added high-concentration fermentation substrate. Therefore, as characteristics of yeast strains for producing high-concentration alcohol, high sugar tolerance and alcohol tolerance are absolute characteristics.

Alcohol resistance can be increased by changing the composition of the fermenter's medium and fermentation conditions, and studies have been conducted to increase alcohol resistance by adding unsaturated fatty acids, proteins, soybean powder, specific vitamins and metal ions to the actual fermenter. come.

However, these attempts have limitations due to economic feasibility and difficulties in application on a large industrial scale. After all, it is advantageous to select and develop alcohol-resistant strains based on the mycological characteristics of yeast itself. In addition, high-concentration sugar is required to produce high-concentration alcohol, and in this case, the death of the strain occurs due to the increase in osmotic pressure. A strain having both excellent sugar tolerance and alcohol tolerance is required. That is, it is necessary to isolate a novel yeast strain having excellent viability in the presence of high concentrations of sugar and alcohol.

Under this background, the inventors of the present inventors, while conducting intensive research to increase the fragrance component of liquor and discover a new strain with excellent sugar tolerance and alcohol resistance, Wickerhamomyces Anomalus KJ-1 strain showed sugar tolerance and alcohol tolerance. The present invention was completed by confirming that this is excellent, and when a liquor is prepared using this, the scent component is enhanced without loss of the scent component to produce a liquor having excellent sensory characteristics.

An object of the present invention is to provide a novel Wickerhamomyces anomalus strain having excellent sugar tolerance, alcohol resistance and fragrance producing ability, and a use thereof.

In order to achieve the above object, the present invention provides a Wickerhamomyces Anomalus KJ-1 strain having excellent sugar tolerance, alcohol resistance and fragrance production ability deposited under accession number KACC 93373P.

In addition, the present invention is an alcoholic beverage manufacturing method comprising the step of culturing Wicker Hamomyces Anomalus KJ-1 strain deposited with Accession No. KACC 93373P in a fermentation medium containing 10 to 50% by weight of glucose to provide.

In addition, the present invention provides a liquor prepared by the above production method.

Wickerhamomyces Anomalus KJ-1 strain of the present invention has excellent sugar tolerance, alcohol resistance and fragrance production ability, so it can be used for producing high-quality alcoholic beverages with improved sensory characteristics, under high concentration sugar conditions or high concentration alcohol conditions. It is possible to grow without dying in

Figure 1 shows the sugar tolerance according to the cell growth rate (%) for each yeast strain.
Figure 2 shows alcohol resistance according to cell growth rate (%) for each yeast strain.
Figure 3 shows the analysis of the main components of the yeast culture solution.
4 shows the distance between yeast culture groups.
5 shows the ABTS radical scavenging ability for each yeast strain.
Figure 6 shows the ACE inhibitory ability of each yeast strain.

Hereinafter, the present invention will be described in detail.

The present invention provides a novel Wickerhamomyces Anomalus KJ-1 strain with excellent sugar tolerance, alcohol resistance and fragrance production ability deposited under accession number KACC 93373P.

The strain may be isolated from traditional yeast, but is not limited thereto.

The strain has excellent glucose tolerance that can grow under the condition that the glucose concentration in the medium is 10 to 50% by weight. The strain has excellent alcohol tolerance that can grow under conditions of an alcohol concentration of 5 to 15% (v/v).

In a specific embodiment of the present invention, anhydrous alcohol was added to YPD (2% glucose, 0.5% yeast extract, 1% bacto-peptone: Sigma-Aldrich, St. Louis, MO, USA) medium at 0, 5, 10, 15 , 20% (v / v) was added, respectively, and after inoculation of the strain, the absorbance at 660 nm was measured after incubation at 25 ° C. for 24 hours.

As a result, it was confirmed that the Wickerhamomyces Anomalus KJ-1 strain of the present invention maintains activity even at a high alcohol concentration of 20% and is a strain with excellent alcohol resistance, and maintains activity even at a high glucose concentration of 50%, thereby reducing sugar tolerance. It was also confirmed that the strain was also excellent.

It can be seen that the Wickerhamomyces Anomalus KJ-1 strain of the present invention is capable of producing alcoholic beverages having better characteristics and sensory taste than conventional strains.

It was confirmed that the Wickerhamomyces Anomalus KJ-1 strain of the present invention has excellent antioxidant activity.

The antioxidant activity of the present invention means that oxygen that enters the body through human breathing has beneficial effects such as making energy necessary for the human body, but in this process, free radicals, which are extra oxygen that is not good for the body, are generated. Oxygen attacks normal cells in the body and acts as a cause of aging or various diseases. Removing this active oxygen is a way to prevent oxidation (aging) of cells, and suppressing such oxidation of cells is antioxidant activity.

In the present invention, in order to confirm the antioxidant activity, the ABTS radical scavenging ability of Wickerhamomyces Anomalus KJ-1 strain was confirmed.

The measurement of the ABTS radical scavenging activity is a universal test method used to measure the antioxidant activity. In the measurement of the antioxidative activity using the ABTS radical, the ABTS free radical generated by the reaction with potassium persulfate is removed by the antioxidant in the extract and is unique to the radical. It is a method using the decolorization of bluish green, which is the color of , to test antioxidant activity, which is one of the defense mechanisms against oxidative stress.

In a specific embodiment of the present invention, as a result of measuring the ABTS radical scavenging ability in order to confirm the antioxidant activity of Wickerhamomyces Anomalus KJ-1 strain, the ABTS radical scavenging ability of Wickerhamomyces Anomalus KJ-1 strain It was confirmed that this was excellent (FIG. 5).

Wickerhamomyces Anomalus KJ-1 strain of the present invention has antihypertensive activity.

The antihypertensive activity may indicate angiotensin converting enzyme (ACE) inhibitory activity.

Since inhibiting the angiotensin converting enzyme (ACE) lowers blood pressure, it will be possible to prevent and treat hypertension.

In a specific embodiment of the present invention, as a result of measuring the ACE inhibitory activity to confirm the antihypertensive activity of Wickerhamomyces anomalus KJ-1 strain, ACE inhibition of Wickerhamomyces anomalus KJ-1 strain It was confirmed that the activity was excellent (FIG. 6).

In addition, the present invention is an alcoholic beverage manufacturing method comprising the step of culturing Wicker Hamomyces Anomalus KJ-1 strain deposited with Accession No. KACC 93373P in a fermentation medium containing 10 to 50% by weight of glucose to provide.

As the raw material, any raw material that can be used for manufacturing alcoholic beverages may be used.

Alcohol (liquor) prepared by preparing a fermentation broth using the Wickerhamomyces anomalus KJ-1 strain of the present invention and distilling and filtering is characterized in that it contains a large amount of excellent flavor components without being lost during the distillation process. .

In addition, the present invention can provide a liquor with increased fragrance components prepared using the Wickerhamomyces Anomalus KJ-1 strain of the present invention.

The liquor may include alcohol flavor, fruit flavor, and sweet flavor, but is not limited thereto.

The alcoholic beverages that can be produced using the Wickerhamomyces Anomalus KJ-1 strain of the present invention include soju, distilled soju, whiskey, brandy, medicinal wine, unsalted rice wine, fermented liquor, beer, Kaoliang liquor, gin, vodka, and It may be one or more selected from the group consisting of wine, but is not limited thereto.

Using the strain of Wickerhamomyces anomalus KJ-1 (Accession No. KACC 93373P) of the present invention, it is possible to prepare alcoholic beverages with excellent sensory characteristics by enhancing fragrance components without loss of fragrance components during production of alcoholic beverages, and high-concentration sugar It is very useful because it can be used to produce alcoholic beverages with enhanced aroma components without being destroyed under conditions or alcohol conditions.

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto.

Example 1. Isolation of novel Wickerhamomyces anomalus KJ-1 strain

As the yeast used in the present invention, Wickerhamomyces anomalus KJ-1 strain, which is a strain that isolates and identifies new resources collected from traditional yeast, and commercially available yeast commonly used in the manufacture of alcoholic beverages were used (Table One).

No. Species Strain KACC no. One Wicherhamomyces anomalus KJ-1 93373P 2 Saccharomyces cerevisiae Lalvin -

Experimental Example 1. Sugar tolerance of Wickerhamomyces Anomalus KJ-1 strain

Glucose tolerant culture medium was prepared by mixing with YPD broth so that the total amount of glucose was 0, 10, 20, 30, 40, or 50%. Inoculate 20 μL each of yeast with the cell concentration adjusted to OD 660 nm = 1 in 180 μL of sugar-tolerant culture medium (YPD broth + glucose), incubate at 25 ° C for 24 hours, and then measure the absorbance of the culture at 660 nm with a spectrophotometer. was measured.

Cell increase was calculated by subtracting the initial absorbance and medium absorbance values from the absorbance value after 24 hours of culture. In addition, the cell growth rate (%) for each concentration was calculated as (calculated cell increase * 100) / 0% cell increase.

The contents of sugar tolerance investigation by yeast strain are shown in Table 2 and FIG. 1. Both Lalvin and KJ-1, control groups, showed a value of 1.0 or more at OD 660nm, which is the increase in glucose up to 30%, and it was found that both strains proliferated the most at 10% (Table 2).

As a result of calculating the growth rate by glucose concentration based on the cell growth rate of 0% glucose treatment, KJ-1 strain showed the best growth rate of 118% compared to the untreated group when treated with 10% glucose, and the growth rate was 42% even when treated with 50% glucose. It was confirmed that the sugar tolerance was high by growing at a low rate without dying (FIG. 1).

Sample 0% 10% 20% 30% 40% 50%. KJ-1 0.97±0.043 1.57±0.023 1.35±0.003 1.00±0.007 0.65±0.014 0.30±0.022 Lalvin 1.07±0.007 1.61±0.006 1.57±0.023 1.31±0.028 0.84±0.064 0.41±0.035 YPD broth 0.05±0.001 0.10±0.002 0.13±0.033 0.12±0.011 0.12±0.003 0.13±0.007

Experimental example 2. Alcohol tolerance of strain Wickerhamomyces anomalus KJ-1

YPD broth was prepared and sterilized under high pressure (121°C, 15 minutes), and when the medium temperature was 40 to 50°C, anhydrous alcohol concentration was added to 0, 5, 10, 15, and 20% (v/v), respectively. . Inoculate 20 μL each of yeast with the cell concentration adjusted to OD 660 nm = 1 in 180 μL of alcohol-added culture medium (YPD + alcohol), seal with parafilm, incubate at 25 ° C for 24 hours, and then measure at 660 nm with a spectrophotometer. The absorbance of the culture medium was measured. Cell increase was calculated by subtracting the initial absorbance and medium absorbance values from the absorbance value after 24 hours of culture.

In addition, cell proliferation rate (%) for each concentration was calculated as (calculated cell increase * 100) / 0% cell increase.

The contents of alcohol resistance investigation by yeast strain are shown in Table 3 and FIG. 2. Both Lalvin, the control group, and KJ-1, the experimental group, showed a value of 1.0 or more at OD 660nm, which is the amount of cell growth up to 5% alcohol concentration, and it was found that both strains proliferated the most at 0% (Table 3). As a result of calculating the growth rate by alcohol concentration based on 0% cell growth, the KJ-1 strain showed the highest growth rate of 99% compared to the untreated group when treated with 5% alcohol, and the growth rate was 14% even when treated with 15%. It was found that it grew at a low rate without alcohol resistance (FIG. 2).

Sample 0% 5% 10% 15% 20% KJ-1 1.04±0.053 1.03±0.018 0.62±0.036 0.13±0.030 0.03±0.002 Lalvin 1.07±0.028 1.02±0.064 0.97±0.047 0.52±0.117 0.03±0.020 YPD broth 0.06±0.003 0.06±0.002 0.06±0.002 0.06±0.002 0.05±0.000

Experimental example 3. Fragrance producing ability of Wickerhamomyces Anomalus KJ-1 strain

3-1. Sniffing test

Yeast was streaked on each YPD Agar medium, and a sniffing test was performed by comparing the strain cultured at 25 ° C for 3 days with the uncultured YPD Agar medium (control). A total of 3 times were performed for each sample.

The results are shown in Table 4. The sensory acceptability was higher in KJ-1.

Sample 1 ^{st 2nd 3rd} KJ-1 alcohol flavor fruit flavor sweet scent Lalvin alcohol flavor pungent scent yeast flavor YPD agar Sikhye incense chestnut scent Que Que Hanhyang

3-2. Electronic nose analysis

For the electronic nose experiment, the vial and the lid sealed with PTFE/silicone were sterilized under high pressure (121℃, 15min), and then inoculated 200 μL each of yeast with the cell concentration adjusted to OD 660nm = 1 in 3 mL of YPD broth medium and cultured for 4 days. So, the sample was prepared. Experimental conditions are shown in Table 5. Electronose analysis (Heracles NEO, Alpha MOS, Toulouse, France) uses C6-C16 alkanes as standards to identify organic compounds by measuring with two polar columns (MXT-5, MXT-1701) and calculating the Kovat index did Data statistical processing was presented by principal component analysis using Alpha MOS software.

Parameter Value Sample quantity 3.2mL in a 20mL vial Type wine Incubation time 20min Incubation temperature 30℃ Injection volume 1,000 µL Trap temperature 30℃ Carrier gas Hydrogen Detector temperature 260℃

The results of electronic nose analysis of the yeast culture medium are shown in FIG. 3 . The contribution rate of the value of the first principal component (PC1) is 97.666%, and the contribution rate of the value of the second principal component (PC2) is 2.334%. Therefore, it can be seen that only the value of the first main component is sufficient information to classify the fragrance pattern. The value of the first principal component is distributed between -10000000 and 20000000, and KJ-1, Lalvin, and YPD are all located in different quadrants, so the volatile content is different between samples. Families were distinguished by independent scent patterns (explaining power 86.057%). YPD, located in the negative direction of the two principal components, was distinguished from other samples and fragrance patterns, but this did not have a high explanatory power of 2.234%.

Therefore, the difference between KJ-1 and the rest of the samples and volatile components will be large. When the distance between the yeast culture groups was calculated, the results were similar to those in Figure 3. The group distance between KJ-1, Lalvin, and YPD was the farthest, and the group distance between Lalvin and YPD was closest (FIG. 4). The volatile compounds of the KJ-1 culture medium measured by the MTX-5 column are shown in Table 6, and the volatile compounds of the Lalvin culture medium are shown in Table 7.

It was found that the volatile compounds of the KJ-1 culture medium contained a sweet flavor as a whole, and had a fruity flavor at the beginning with RTs of 13.07 to 43.67 (Table 6). This is very similar to the results of the sniffing test, so it is data that objectively supports the sensory characteristics (Table 5). In the case of Lalvin's culture medium, the RT is 14.29~34.24, and it has a strong alcohol scent at the beginning, a fruity scent toward the middle, and a floral scent toward the latter half.

Retention
time Formula Name Relevance
index Sensory descriptors 13.07 C ₃ H ₉ N Trimethylamine 92.83 Amine;Ammoniacal;Fishy;Fruity;Oily;Pungent;Rancid;Sweaty 15.42 C ₃ H ₆ O Propanal 94.27 acetaldehyde;Cocoa;Earthy;Etheral;Nutty;Plastic;Pungent;Solvent 23.06 C ₄ H ₈ O ₂ Methyl propanoate 97.17 Apple; Etheral; Fresh; Fruit; Harsh; Rum; Strawberry; Sweet 26.81 C ₄ H ₁₀ O n-butanol 90.61 Alcoholic;amyl alcohol;Banana;Cheese;Fermented;Fruity;Fusel;Harsh;Medicinal;Oil;Rancid;Strong;Sweet 34.36 C ₅ H ₁₂ O S(-)2-methyl-1-butanol 91.59 Fresh;Fruity;Malty 43.67 C ₆ H ₁₂ O ₂ Butyl acetate 98.61 Banana;Bitter;Etheral;Fruity;Green;Pear;Pineapple;Pleasant;Solvent;Strong;Sweaty;Sweet 50.22 C ₇ H ₁₄ O ₂ isoamyl acetate 99.27 Apple;Banana;Ester;Fresh;Fruity;fruity (sweet);Pear;Pleasant;Solvent;Sweet 58.25 C ₂ H ₆ S ₃ Dimethyl trisulfide 96.68 Alliaceous; Cabbage; Cauliflower; Fishy; Meaty; Onion; onion (cooked); Rotten food; Sulfurous 64.71 C ₅ H ₆ O ₃ 4-hydroxy-5-methyl-3(2H)-furanone 96.56 Balsamic;Candy;Caramelized;Cotton candy;Sweet 77.39 C ₁₀ H ₁₄ O ()-Carvone 92.54 basil;Bitter;Caraway;Fennel;Minty;Peppermint;Speaarmint;Sweet

Retention
time Formula Name Relevance
index Sensory descriptors 14.29 _CH4O Methanol 95.51 Alcoholic; Pungent; Strong 15.38 C ₃ H ₄ O Propenal 96.04 Almond; Cherry; choking 16.43 C ₅ H ₁₂ Pentane 92.92 Alkanes; Gasoline 21.31 C ₄ H ₁₀ O 2-butanol 97.7 Pleasant; Strong; Sweet; Wine 22.42 C ₂ H ₂ C _l2 Ethene, 1,2-dichloro, (Z)- 98.43 Sweet 34.24 C ₅ H ₁₂ O 3-Methyl-1-butanol 95.06 Alcoholic; Balsamic; Bitter; Burnt; Cheese; Fermented; Fruity; Harsh; Malty; onion (ripe); Pungent; unpleasant; Whisky 38.95 C ₅ H ₁₁ C _l 1-Chloropentane 92.77 Green plant; Sweet 50.17 C ₇ H ₁₄ O ₂ isoamyl acetate 99.11 Apple;Banana;Ester;Fresh;Fruity;fruity (sweet);Pear;Pleasant;Solvent;Sweet 58.59 C ₁₀ H ₁₆ sabinene 98.92 Citrus; Fresh; Pepper; Pine; Spicy; Sweet; Turpentine; Woody 59.51 C ₈ H ₁₆ O 3-Octanone 98.18 Butter;Fresh;Fruity;Herbaceous;Lavender;Mild;Mushroom;Resinous;Sweet 60.43 C ₁₀ H ₂₂ Decane 95.59 Alkane;Fruity;Fusel;Sweet 73.21 C ₁₁ H ₂₀ O 2-Methylisoborneol 99.1 Earthy; Musty 73.21 C ₁₀ H ₂₀ O ₂ propionic acid heptyl ester 98.93 Apricot;Floral;Rose 83.63 C ₁₂ H ₂₄ O ₂ Propyl nonanoate 94.83 Fermented; Melon 86.21 C ₁₂ H ₂₂ O ₂ octyl crotonate 97.1 Chemical;Fruity;Harsh;Hay;Mushroom;Winey 95.85 C ₁₅ H ₃₀ O ₂ Nonanoic acid, hexyl ester 93.84 Brandy;Floral;Fruity;Green;Vegetable

Experimental Example 4. Antioxidant activity of Wickerhamomyces Anomalus KJ-1 strain

ABTS radical scavenging activity was measured using the ABTS Antioxidant Kit (Zen-Bio). After dispensing 10 μL samples and standards into 96 well plates, 10 μL of assay buffer was added to each well as a negative control. 20 μL/well myoglobin working solution was dispensed. 100 μL of ABTS solution was added to each well, and placed on a plate shaker at room temperature of 25° C. for 5 minutes. To stop the reaction, 50 μL of stop solution was added to each well and absorbance was measured at 405 nm. For comparison of activity, the positive control group used Trolox solution, and the result value was calculated by converting Trolox to 100%.

Antioxidant activity measurement using ABTS radical is one of the defense mechanisms against oxidative stress by using the fact that the ABTS free radical generated by the reaction with potassium persulfate is removed by the antioxidant substance in the extract and the blue-green color unique to the radical is decolored. It is used as a method for assaying phosphorus antioxidant activity. ABTS radical scavenging activity was measured using the ABTS Antioxidant Kit (Zen Bio), and as a result of converting Trolox, a positive control group, to 100%, Lalvin was 100.6% and KJ-1 was 128.8%, which was 128% more active than commercially available yeast. It can be confirmed that it is high (FIG. 5).

Through this, it shows that the antioxidant activity of Wickerhamomyces Anomalus KJ-1 strain is excellent.

Experimental Example 5. Antihypertensive activity of Wickerhamomyces Anomalus KJ-1 strain

For ACE inhibition screening, it was measured using a kit (Kamiya biomedical company, KT-534). Yeast is pre-cultured on YPD agar medium (25℃, 2~3 days), and 1 colony is inoculated into YPD broth, then cultured at 25℃ for 24~72 hours, and the culture medium with the concentration adjusted to OD 660nm=1 is added to 5 mL YPD broth The medium was inoculated with 2%, cultured for 7 days, and the culture medium was centrifuged, and the supernatant was used in the experiment. After putting 20 μL of the sample, mixing equal amounts of substrate buffer and enzyme working solution, reacting at 37 ° C for 1 hour, adding 200 μL of Indidcator wirking solution and incubating at room temperature for 10 minutes. As a control, distilled water was used instead of the sample, and absorbance at 450 nm was measured using Captopril (1%) as a positive control. ACE inhibitory activity was calculated by the following formula and expressed as a percentage (%).

ACE inhibition (%) = [(A _{blank 1} - A _sample )/(A _{blank 1} - A _blank2 )] X 100

ACE inhibitors inhibit the production of angiotensin-converting enzyme, thereby relaxing blood vessels and lowering blood pressure. As a result of analysis using 1% Captopril as a standard material as a positive control to confirm ACE inhibition, the commercially available yeast Lalvin was 51.2% and KJ-1 was 61.6%, showing 120% higher activity than commercially available yeast. Yes (FIG. 6).

Through this, it shows that the antihypertensive activity of Wickerhamomyces Anomalus KJ-1 strain is excellent.

Agricultural Biotechnology Research Institute KACC93373P 20211012

Claims (12)

Wicker Hamomyces Anomalus KJ-1 strain deposited under Accession No. KACC 93373P.
According to claim 1,
The strain is characterized in that it has excellent glucose tolerance that can grow under conditions where the glucose concentration in the medium is 10 to 50% by weight, Wickerhamomyces Anomalus KJ-1 strain.
According to claim 1,
The strain is Wicker Hamomyces Anomalus KJ-1 strain, characterized in that it has excellent alcohol resistance that can grow under conditions of an alcohol concentration of 5 to 15% (v / v).
According to claim 1,
The strain is Wicker Hamomyces Anomalus KJ-1 strain, characterized in that it has excellent antioxidant activity.
According to claim 4,
Wickerhamomyces anomalus KJ-1 strain, characterized in that the antioxidant activity exhibits ABTS radical scavenging activity.
According to claim 1,
The strain is Wickerhamomyces Anomalus KJ-1 strain, characterized in that the antihypertensive activity is excellent.
According to claim 6,
The antihypertensive activity is Wickerhamomyces Anomalus KJ-1 strain, characterized in that exhibits angiotensin converting enzyme (ACE) inhibitory activity.
A method for producing alcoholic beverages comprising the step of culturing Wicker Hamomyces Anomalus KJ-1 strain deposited under Accession No. KACC 93373P in a fermentation medium containing 10 to 50% by weight of glucose.
According to claim 8,
The alcoholic beverage manufacturing method, characterized in that it comprises an alcohol flavor, fruit flavor and sweet flavor.
Liquor produced by the manufacturing method of claim 8.
According to claim 10,
The liquor is characterized in that the fragrance component is increased, the liquor.
According to claim 10,
The liquor is characterized in that at least one selected from the group consisting of soju, distilled soju, whiskey, brandy, yakju, takju, fermented liquor, beer, kaoliangju, gin, vodka and wine.

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