KR20210073175A - Enzymatically treated stevia sweetener prepared from single steviol glycosides of particular ratio and manufacturing method of alcoholic liquors using the same - Google Patents

Abstract

The present invention relates to an enzymatically-treated stevia sweetener prepared by using single steviol glycosides of a specific ratio and a method of preparing liquor using the same, and more specifically, to an enzymatically-treated stevia sweetener, which is obtained by glycosylating a stevia glycoside composition in which dulcoside A, stevioside, rebaudioside C and rebaudioside A are contained in a specific ratio, and a method of preparing diluted soju prepared by adding the enzymatically-treated stevia sweetener so as to have improved sweetness and bitterness, harmonious and soft tastes and a clean tailing flavor. According to the present invention, when the enzymatically-treated stevia sweetener is added to prepare diluted soju, not only the overall preference but also preferences in terms of detailed property aspects of soju such as softness, cleanliness, and properties of soju itself can be improved. In addition, it is possible to prepare diluted soju having uniform quality without the influence of a sweetness which can subtly vary according to a variety or a crop situation of stevia leaves.

Description

Enzymatically treated stevia sweetener prepared from single steviol glycosides of particular ratio and manufacturing method of alcoholic liquors using the same

The present invention relates to an enzyme-treated stevia sweetener prepared from a single steviol glycoside in a specific ratio, and a method for preparing alcoholic beverages using the same. More specifically, glycosylation of a steviol glycoside composition comprising four steviol glycoside single components, dulcoside A, stevioside, rebaudioside C and rebaudioside A, isolated from stevia leaf extract in a specific ratio The present invention relates to an enzyme-treated stevia sweetener obtained by doing so, and a method for preparing a harmonious diluted soju with smooth, clean, sweet and bitter taste by adding the same.

Conventional soju was mostly diluted soju with alcohol content of 20% by volume (vol%), but recently, alcohol products with an alcohol content of 16.5 to 17.0% by volume are positioned as the main products due to the low level of alcohol.

In the case of highly diluted soju, the sweetness and bitterness of alcohol from alcohol harmonize and masking the taste is the most important in product development. It has been using natural sweeteners such as enzyme-treated stevia, and has been manufacturing diluted soju with products that constantly improve sweetness.

The steviol glycoside and enzyme-treated stevia are generally characterized by sensory elements such as bitter taste, after-sweetness, high sweetness, and astringent flavor, and stevioside, a steviol glycoside produced by extraction from natural stevia leaves. ST) has strong sweetness but also strong bitterness, which is why it is unfavorable for sensory evaluation. In the case of rebaudioside A (RA), the sweetness is good and the bitterness is lower than ST, but it is expensive when sold. In addition, steviol glycosides including rebaudioside C (RC) and dulcoside A (dulcoside A, DA) are less than 5% by weight of the total steviol glycosides in the natural stevia extract, which makes it difficult to extract with a single extract. Manufactured at a high price due to time-consuming manufacturing process and equipment investment, etc., the steviol glycoside that is most used in food and beverages is not commercialized except for RA.

On the other hand, there is a problem in that the composition of steviol glycosides is different depending on the crop or variety of stevia, so there is a problem in showing subtle differences in sweetness and taste like soju in diluted soju.

Korean Patent Publication No. 10-2019-0094151 Japanese Patent Laid-Open No. 2018-121534

Under this background, the present inventors produced shochu, which is organoleptically improved compared to commercially available soju products containing steviol glycoside, and at the same time, glycosylated a composition containing four types of single steviol glycoside in a specific ratio as a sweetener for shochu production. By using the prepared enzyme-treated stevia sweetener, it was confirmed that dilution-type soju of uniform quality can be produced without affecting the sweetness, which may be different depending on the variety or crop of stevia leaves, thereby completing the present invention.

An object of the present invention is to glycosylate a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A; It is to provide a method for producing an enzyme-treated stevia sweetener comprising a.

Another object of the present invention is 0 to 3% by weight of dulcoside A; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; and 35 to 45% by weight of rebaudioside A; To provide an enzyme-treated stevia sweetener prepared by glycosylation of the included steviol glycoside composition.

Another object of the present invention is to obtain an enzyme-treated stevia sweetener by a) glycosylating a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A; b) adding the enzyme-treated stevia sweetener obtained in step a) to alcohol having an alcohol concentration of 50 vol%; And c) after purifying and filtering the alcohol of step b), the step of fermenting; to provide a method for producing dilute soju, including a.

Another object of the present invention is to provide a diluted soju containing 0.01 to 0.04% by weight of the enzyme-treated stevia sweetener.

The object of the present invention is not limited to the object mentioned above. The objects of the present invention will become more apparent from the following description, and will be realized by means and combinations thereof described in the claims.

In order to achieve the above object, the following solutions are provided.

In one aspect of the present invention, a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A is glycosylated. It provides a method for producing an enzyme-treated stevia sweetener, comprising the step.

In one aspect of the present invention, the steviol glycoside composition is 0 to 3% by weight of dulcoside A based on the total weight of the composition; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; And rebaudioside A is included in 35 to 45% by weight, it provides a method for producing an enzyme-treated stevia sweetener.

Another aspect of the present invention is 0 to 3% by weight of dulcoside A; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; and 35 to 45% by weight of rebaudioside A; Provided is an enzyme-treated stevia sweetener prepared by glycosylation of the included steviol glycoside composition.

In another aspect of the present invention, the enzyme-treated stevia sweetener includes steviol glycoside G1, stevioside, rebaudioside C, rebaudioside A, and steviol glycoside G2. .

Another aspect of the present invention is a method comprising: a) glycosylating a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A to obtain an enzyme-treated stevia sweetener; b) adding the enzyme-treated stevia sweetener obtained in step a) to alcohol having an alcohol concentration of 50 vol%; And c) after purifying and filtering the alcohol of step b), it provides a method for preparing dilute soju, including a.

In another aspect of the present invention, the steviol glycoside composition of step a) contains 0 to 3% by weight of dulcoside A based on the total weight of the composition; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; And rebaudioside A is included in 35 to 45% by weight, it provides a method for producing dilution type soju.

In another aspect of the present invention, the enzyme-treated stevia sweetener in step b) provides a method for preparing diluted soju, which is added in an amount of 0.01 to 0.04% by weight based on the total weight of the soju.

In another aspect of the present invention, the number of step c) is adjusted to be 15 to 17 vol% of alcohol concentration, it provides a method for producing dilute soju.

In another aspect of the present invention, the diluted soju, based on the total weight of soju, steviol glycoside G1 is 0.0002 to 0.0010% by weight; 0.0025 to 0.0060% by weight of stevioside; 0.0001 to 0.0005% by weight of rebaudioside C; 0.0055 to 0.02% by weight of rebaudioside A; and 0.003 to 0.006 wt% of steviol glycoside G2; It provides a method for preparing diluted soju, including.

Another aspect of the present invention provides a diluted soju containing 0.01 to 0.04% by weight of the enzyme-treated stevia sweetener.

Glycosylation of a steviol glycoside composition comprising a single component of steviol glycoside, dulcoside A, stevioside, rebaudioside C, and rebaudioside A, isolated from a stevia leaf extract according to an aspect of the present invention in a specific ratio When the enzyme-treated stevia sweetener obtained by the above method is added during the preparation of diluted soju, it is possible to improve not only overall preference, but also in terms of detailed properties of soju, such as softness, cleanliness, and the quality of soju.

In addition, the enzyme-treated stevia sweetener prepared with a single steviol glycoside in a specific ratio according to one aspect of the present invention is capable of producing diluted soju of uniform quality without affecting the sweetening quality that may be subtly different depending on the variety or crop of stevia leaves. It has the effect of making it possible.

On the other hand, in the prior art, there is a problem that it is difficult to manage the subtle difference of high sweetener, which is the main additive, for each lot during the manufacture of soju, so that the difference identification has to depend only on the sensory evaluation result. When an enzyme-treated stevia sweetener prepared from a single steviol glycoside is applied to the production of soju, the flavor of soju products can be stably managed by adding a standardized sweetener, and quality control is also easy.

The effects of the present invention are not limited to the above-mentioned effects. It should be understood that the effects of the present invention include all effects that can be inferred from the following description.

1 is a process diagram schematically illustrating the manufacturing process of diluted soju using an enzyme-treated stevia sweetener prepared with a single steviol glycoside in a specific ratio according to the present invention.

The above objects, other objects, features and advantages of the present invention will be easily understood through the following preferred embodiments in conjunction with the accompanying drawings. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the embodiments introduced herein are provided so that the disclosed content may be thorough and complete, and the spirit of the present invention may be sufficiently conveyed to those skilled in the art.

In the present specification, terms such as “comprise” or “have” are intended to designate that a feature, number, step, operation, component, part, or combination thereof described in the specification exists, but one or more other features It is to be understood that it does not preclude the possibility of the presence or addition of numbers, steps, operations, components, parts, or combinations thereof. Also, when a part of a layer, film, region, plate, etc. is said to be "on" another part, this includes not only the case where it is "directly on" another part, but also the case where another part is in the middle. Conversely, when a part of a layer, film, region, plate, etc. is said to be “under” another part, it includes not only cases where it is “directly under” another part, but also cases where another part is in the middle.

In this specification, when a range is described for a variable, the variable will be understood to include all values within the stated range including the stated endpoints of the range. For example, a range of “5 to 10” includes the values of 5, 6, 7, 8, 9, and 10, as well as any subranges such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, etc. It will be understood to include any value between integers that are appropriate for the scope of the recited range, such as 5.5, 6.5, 7.5, 5.5 to 8.5 and 6.5 to 9, and the like. Also for example, a range of "10% to 30%" includes values such as 10%, 11%, 12%, 13%, and all integers up to and including 30%, as well as 10% to 15%, 12% to It will be understood to include any subrange, such as 18%, 20% to 30%, etc., as well as any value between reasonable integers within the scope of the recited range, such as 10.5%, 15.5%, 25.5%, and the like.

Hereinafter, the present invention will be specifically described.

The present invention relates to an enzyme-treated stevia sweetener prepared from a single steviol glycoside in a specific ratio, and a method for preparing alcoholic beverages using the same. More specifically, an enzyme-treated stevia sweetener obtained by glycosylation of a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A in a specific ratio and improved by adding the same It relates to a method for manufacturing diluted soju with sweet and bitter qualities, harmonious, soft, and clean aftertaste.

In one aspect of the present invention, a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A is glycosylated. It provides a method for producing an enzyme-treated stevia sweetener, comprising the step.

In the present invention, the steviol glycoside composition is not limited thereto, but based on the total weight of the composition, 0 to 3% by weight of dulcoside A, a single component of steviol glycoside isolated from stevia leaf extract; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; and 35 to 45% by weight of rebaudioside A; may be included. preferably 0 to 2% by weight of dulcoside A; stevioside in an amount of 48 to 52% by weight; 2 to 5% by weight of rebaudioside C; and 38 to 42% by weight of rebaudioside A; may be included. more preferably 0% or 1% by weight of dulcoside A; Stevioside 50% by weight; 2% or 5% by weight of rebaudioside C; and 40% by weight of rebaudioside A; may be included.

In one embodiment of the present invention, the steviol glycoside composition is dulcoside as the main single component of the steviol glycoside isolated from a stevia extract obtained by hot water extraction of dry leaf powder of Stevia rebaudiana Bertoni. A (dulcoside A, DA), stevioside (ST), rebaudioside C (rebaudioside C, RC) and rebaudioside A (rebaudioside A, RA) are included, and other steviol glycosides are single As ingredients steviobioside (STB), rubusoside (RuB), rebaudioside B (RB), rebaudioside D (rebaudioside C, RD), and rebaudioside F (rebaudioside F, RF) and the like were also confirmed to be included (see Examples 1-4).

In the present invention, the glycosylation (glycosylation) is a reaction of enzymatic transfer from a compound having a glycosyl group to another compound, but is not limited thereto, and can be performed using cyclodextrin glucanotransferase (CGTase). have.

In one embodiment of the present invention, a steviol glycoside comprising a single component of steviol glycoside isolated from stevia leaf extract, dulcoside A, stevioside, rebaudioside C, and four types of rebaudioside A in a specific ratio After mixing the composition with dextrin in a weight ratio of 1:3 and dissolving it in water three times the total weight of the raw material, cyclodextrin glucanotransferase (CGTase) is added to the enzymatic reaction, and the enzyme standardized as a sweetener for soju Treated stevia was obtained (see Examples 1-5).

Another aspect of the present invention is 0 to 3% by weight of dulcoside A; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; and 35 to 45% by weight of rebaudioside A; Provided is an enzyme-treated stevia sweetener prepared by glycosylation of the included steviol glycoside composition.

In the present invention, the enzyme-treated stevia sweetener is not limited thereto, but may include steviol glycoside G1, stevioside, rebaudioside C, rebaudioside A, and steviol glycoside G2.

The steviol glycoside composition and glycosylation are as described above.

In one embodiment of the present invention, after glycosylation of the steviol glycoside composition combined in a specific ratio, HPLC analysis results show that dulcoside A (DA), stevioside (ST), rebaudioside C (RC), rebau Dioside A (RA) and other components (STB, RuB, RB, RD, RF, etc.) are steviol glycoside G1 (G1) and steviol glycoside G2 (G2) and It was confirmed that the same steviol glycoside was newly generated and lost in the case of DA (see Examples 1-5).

Another aspect of the present invention is a method comprising: a) glycosylating a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A to obtain an enzyme-treated stevia sweetener; b) adding the enzyme-treated stevia sweetener obtained in step a) to alcohol having an alcohol concentration of 50 vol%; And c) after purifying and filtering the alcohol of step b), it provides a method for preparing dilute soju, including a.

The manufacturing method of the diluted soju is described in more detail step by step as follows.

Step a) is a step of preparing an enzyme-treated stevia sweetener. Specifically, by glycosylation of a steviol glycoside composition comprising a single component of steviol glycoside isolated from stevia leaf extract, dulcoside A, stevioside, rebaudioside C, and four types of rebaudioside A in a specific ratio This is a step to obtain enzyme-treated stevia sweetener.

Although not limited thereto, the steviol glycoside composition may contain 0 to 3% by weight of dulcoside A based on the total weight of the composition; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; and 35 to 45% by weight of rebaudioside A; may be included. preferably 0 to 2% by weight of dulcoside A; stevioside in an amount of 48 to 52% by weight; 2 to 5% by weight of rebaudioside C; and 38 to 42% by weight of rebaudioside A; may be included. more preferably 0% or 1% by weight of dulcoside A; Stevioside 50% by weight; 2% or 5% by weight of rebaudioside C; and 40% by weight of rebaudioside A; may be included.

Step b) is a step of adding the enzyme-treated stevia sweetener obtained in step a) to alcohol. Specifically, it is a step of adding the enzyme-treated stevia sweetener to an alcohol having an alcohol concentration of 50 vol% diluted by mixing an alcohol having an alcohol concentration of 95 vol% with suitable water based on drinking water.

In the present invention, suitable water for drinking water standards is not limited thereto, but may be groundwater, drinking spring water, natural water, reverse osmosis water (RO water), alkaline water, and the like.

The alcohol is not limited thereto, but may be neutral alcohol or grain alcohol. The alcohol refers to an alcohol prepared by fermenting starchy raw materials (grains, documents, etc.) and then refining the flavor derived from the raw material through continuous distillation so that only alcohol, which is ethanol, remains.

In order to deodorize the alcohol, but not limited thereto, 0.01 to 0.2 wt%, 0.03 to 0.1 wt%, or 0.05 wt% of activated carbon may be added based on the total weight of the alcohol.

The deodorization is not limited thereto, but may be performed by stirring for 1 to 5 hours, 2 to 4 hours, or 3 hours after the addition of activated carbon to alcohol.

Activated carbon is separated from the deodorized alcohol through ultra-fine filtration, and finally alcohol diluted with an alcohol concentration of 50 vol% can be obtained.

The ultra-fine filtration is not limited thereto, but may be performed using a filtration membrane having a pore size of 0.2 to 13 μm, 0.5 to 10 μm, 1 to 8 μm, or 5 μm.

The enzyme-treated stevia sweetener in step b) is not limited thereto, but may be added in an amount of 0.01 to 0.04% by weight, 0.01 to 0.03% by weight, or 0.02% by weight based on the total weight of soju.

Meanwhile, although not limited thereto, in addition to the enzyme-treated stevia sweetener, one or more additives selected from the group of sugar, glucose, fructose, citric acid, amino acids, sorbitol, erythritol, tomatine, and inorganic salts according to the preference of the soju to be finally prepared may be additionally added. In addition, functional food extracts may be added.

Step c) is a step of purifying and filtering the diluted alcohol (alcohol concentration of 50 vol%) to which the enzyme-treated stevia sweetener is added in step b), and then adjusting it to the target final alcohol concentration, that is, performing it.

The diluted alcohol to which the enzyme-treated stevia sweetener is added is not limited thereto, but may be purified and filtered using an ultra-fine filtration membrane.

The filtration membrane is not limited thereto, but may have a pore size of 0.2 to 13 μm, 0.5 to 10 μm, 1 to 8 μm, or 5 μm.

Although not limited thereto, the dilute alcohol to which the enzyme-treated stevia sweetener is added is subjected to an additional process capable of deodorizing and decolorizing with cotton and limestone, activated carbon, diatomaceous earth, ion exchange resin, special carbon, etc. before purification and filtration. can Accordingly, it is possible to improve the flavor of the diluted soju finally obtained by filtration with an ultra-fine filtration membrane.

The number is not limited thereto, but may be adjusted so that the alcohol concentration is 15 to 17 vol%, or 16.5 vol%.

By going through the above water process, it is possible to obtain a diluted soju with a final alcohol concentration of 15 to 17 vol%, or 16.5 vol%.

The obtained diluted soju is not limited thereto, but based on the total weight of soju, steviol glycoside G1 is 0.0002 to 0.0010% by weight; 0.0025 to 0.0060% by weight of stevioside; 0.0001 to 0.0005% by weight of rebaudioside C; 0.0055 to 0.02% by weight of rebaudioside A; and 0.003 to 0.006 wt% of steviol glycoside G2; may be included.

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example 1: Preparation of enzyme-treated stevia sweetener with a specific ratio of single steviol glycoside

Example 1-1: Stevia leaf extract containing crude steviol glycoside was obtained

A stevia leaf extract containing 95% by weight or more of crude steviol glycosides was obtained. Specifically, demineralized water was injected in the form of steam to the dried leaf powder of Stevia rebaudiana Bertoni, and heated at 70° C. for 2 hours for hot water extraction. The precipitate contained in the stevia leaf extract obtained as described above was removed through filtration and washed with water. The washed stevia leaf extract was passed through a cation exchange resin to obtain a stevia leaf extract concentrate containing crude steviol glycosides containing sweetening components such as stevioside and rebaudioside.

Example 1-2: Pure Total Steviol Glycoside obtained by Purification of Stevia Leaf Extract

Coloring substances and impurities from the stevia leaf extract concentrate containing the crude steviol glycoside obtained in Example 1-1 were removed to obtain a high-purity total steviol glycoside. Specifically, aluminum sulfate (Al ₂ (SO ₄ ) ₃ ), activated carbon, lime milk (Ca(OH) ₂ ), and sodium hydroxide (NaOH) were added to the stevia leaf extract to precipitate and agglomerate a significant amount of impurities and colored substances. was passed through an ultrafiltration membrane to remove impurities of low molecular weight, and a steviol glycoside concentrate containing a steviol glycoside and a large amount of chlorophyll was obtained.

In order to remove a large amount of chlorophyll contained in the steviol glycoside concentrate, the steviol glycoside concentrate is mixed with an organic solvent such as benzene, butyl alcohol, isopropyl alcohol, and n-hexane to remove a large amount of chlorophyll. A glycoside was obtained.

Example 1-3: Isolation of single component steviol glycoside

The high-purity total steviol glycoside obtained in Example 1-2 was separated into a single component steviol glycoside using an adsorption resin. Specifically, the total steviol glycoside concentrate was passed through a column filled with an adsorption resin to adsorb the steviol glycoside. After adsorption, it was sufficiently washed and eluted with methanol to obtain an eluent containing a steviol glycoside. After passing the eluent obtained above using a column filled with an ion exchange resin, activated carbon was added to the flow-through solution and stirred. The stirred solution was filtered, concentrated and dried to obtain a mixture of steviol glycosides containing sweetening components such as stevioside and rebaudioside.

Ethanol, methanol, and water were mixed with the obtained steviol glycoside mixture, heated to reflux for 10 minutes, and cooled to 22° C. for 16 hours. Then, it was washed with a mixture of ethanol and methanol, and dried under reduced pressure in a vacuum oven at 50° C. for 18 hours to obtain a final purified total steviol glycoside.

In order to analyze the single component of the obtained total steviol glycosides, the total steviol glycosides were dissolved to 5% by weight based on the total weight of water to prepare 5 Brix. A single component steviol glycoside extract was obtained by adsorbing the total steviol glycoside prepared as described above to a column filled with 200 ml of adsorption resin (XAD-7), and eluting with 200 ml of ethanol (70% by volume). obtained. At this time, the flow-through solution eluted from the column is passed by adjusting at a rate of 1 ml/min, and it is sampled at 10 ml intervals and analyzed by HPLC to confirm retention time (RT) and peak area (area [%]). did.

Specifically, the eluted single component steviol glycoside extract is dulcoside A (dulcoside A, DA), stevioside (ST), rebaudioside C (rebaudioside C, RC) and rebaudioside A, respectively. (rebaudioside A, RA) was analyzed to be the major single component. In addition, as a single component of steviol glycoside, steviobioside (STB), rubusoside (rubusoside, RuB), rebaudioside B (rebaudioside B, RB), rebaudioside D (rebaudioside C, RD) , and rebaudioside F (RF).

Then, the eluted steviol glycoside extract was concentrated as it is to a single composition of steviol glycoside, or impurities were removed with a cation exchange resin, an anion exchange resin, activated carbon, etc. if necessary. In addition, the sweetener was re-adsorbed on the adsorption resin, eluted with ethanol (70% by volume), and the eluent was concentrated and dried, and the extract was concentrated by using an extract or other conventional purification means such as turbidity.

Example 1-4: Establishment of the composition of a single steviol glycoside in a specific ratio

Steviol as the optimal sweetener for soju by selecting a specific ratio of steviol glycosides as a single ingredient in order to minimize the effect of sweetening quality, which may vary depending on the stevia leaf variety or crop, and to manufacture a soju product with easy quality control. The glycoside composition was established.

Specifically, a single component steviol glycoside isolated in Examples 1-3, specifically dulcoside A (DA), stevioside (ST), rebaudioside C (RC), rebaudioside A (RA) ) and other ingredients (steviobioside (STB), rubusoside (RuB), rebaudioside B (RB), rebaudioside D (RD), and rebaudioside F (RF)) are listed in the table below. A steviol glycoside composition was constructed by combining them in a ratio of 1 (based on weight %).

test sample DA ST RC RA other ingredients T-1 0 50 2 40 8 T-2 0 60 2 30 8 T-3 One 50 5 40 4 T-4 3 60 5 30 2

Example 1-5: Preparation of enzyme-treated stevia sweetener by glycosylation of a single steviol glycoside in a specific ratio

Each of the single steviol glycoside compositions (T-1 to T-4 samples) in a specific ratio selected in Examples 1-4 were mixed with dextrin in a weight ratio of 1:3 and dissolved in water three times the total weight of the raw material, 0.06 wt% of cyclodextrin glucanotransferase (CGTase) was added based on the total weight, followed by enzymatic reaction in a thermostat at 65°C for 15 hours. Thereafter, it was cooled at room temperature (15 to 25° C.), and the enzyme action was inactivated for 2 hours in a thermostat at 95° C. The inactivated enzyme reaction solution was filtered and separated, concentrated and dried to finally glycosylate a single steviol glycoside in a specific ratio to obtain enzyme-treated stevia as a sweetener for soju.

The components of the enzyme-treated stevia sweetener finally obtained by glycosylation of a single steviol glycoside in the specified ratio were confirmed through HPLC analysis (conditions; UV 210 nm, column μ-Bondapak C18 (3.9×30 cm, I.D)). The results are shown in Table 2 below. On the other hand, the retention time (RT) of each analyte component was shown in Table 3 below.

Specifically, as a result of HPLC analysis, steviol glycoside in which dulcoside A (DA), stevioside (ST), rebaudioside C (RC), rebaudioside A (RA) and other components are combined in a specific ratio is an enzyme Through the treatment, steviol glycosides such as steviol glycoside G1 (G1) and steviol glycoside G2 (G2) are newly generated by the addition of glucose, that is, glycosylation, and it was found that in the case of DA, it was lost.

test sample G1 DA ST RC RA G2 other ingredients T-1 2 0 16 One 55 21 5 T-2 2 0 25 One 30 26 16 T-3 4 0 17 2 55 19 3 T-4 4 0 27 2 32 25 10 Commercially available products containing steviol glycosides 12 One 33 2 40 10 2

* Unit: Area% (based on 100%)

* Other ingredients: Stevioside (STB), rubusoside (RuB), rebaudioside B (RB), rebaudioside D (RD), and steviol glycosides such as rebaudioside F (RF) , dextrin, water, etc.

Enzyme-treated stevia single ingredient RT (min) Steviol glycoside G1 (G1) 7.4 Dulcoside A (DA) 7.8 Stevioside (ST) 9.0 Rebaudioside C (RC) 10.4 Rebaudioside A (RA) 14.0 Steviol glycoside G2 (G2) 16.8

* Retention time (RT): Time to measure Area value during HPLC analysis

* RT for other ingredients: time other than 7-17 minutes

Example 2: Preparation of diluted soju using enzyme-treated stevia sweetener prepared with a single steviol glycoside in a specific ratio

A diluted soju was prepared by adding the enzyme-treated stevia sweetener finally obtained in Examples 1-5.

Specifically, it was diluted to 50% by volume of alcohol by mixing 95% by volume of alcohol with suitable water based on drinking water (alcohol:water = 52.6:47.4 (v%:v%)). Activated carbon in an amount of 0.05 wt % based on the total weight was added to the diluted alcohol, stirred for 3 hours, stopped for 30 minutes, and activated carbon and diluted alcohol were separated through ultra-fine filtration. In this case, ultra-fine filtration was performed using an ultra-fine filtration membrane having a pore size of 5.0 μm. Based on the total weight of the diluted alcohol of 50% by volume of the filtered alcohol, 0.02% by weight of the enzyme-treated stevia sweetener finally obtained in Examples 1-5, 0.1% by weight of fructose and 0.05% by weight of erythritol as additional sweetening additives was added. After that, the final alcohol concentration was adjusted to 16.5% by volume (that is, the alcohol content was adjusted to 16.5), that is, after a water process, filtration was performed with an ultra-fine filtration membrane having a 5.0 μm pore size to prepare diluted soju.

The diluted soju composition is shown in Table 4 below.

Furtherance Test sample (unit: % by weight) T-1 T-2 T-3 T-4 Commercially available products containing steviol glycosides spirits
(34.00 wt%) 50% by volume
diluted spirits 34.0000 34.0000 34.0000 34.0000 34.0000 water
(65.83 wt%) water 65.8300 65.8300 65.8300 65.8300 65.8300 Enzyme-treated Stevia Sweetener
(0.02
weight%) G1 0.0004 0.0004 0.0008 0.0008 0.0024 DA 0.0000 0.0000 0.0000 0.0000 0.0002 ST 0.0032 0.0050 0.0034 0.0054 0.0066 RC 0.0002 0.0002 0.0004 0.0004 0.0004 RA 0.0110 0.0060 0.0110 0.0064 0.0080 G2 0.0042 0.0052 0.0038 0.0050 0.0020 other ingredients 0.0010 0.0032 0.0006 0.0020 0.0004 sweetness
additive
(0.15
weight%) fruit sugar 0.1000 0.1000 0.1000 0.1000 0.1000 erythritol 0.0500 0.0500 0.0500 0.0500 0.0500 total amount 100.0000 100.0000 100.0000 100.0000 100.0000

* Other ingredients: Stevioside (STB), rubusoside (RuB), rebaudioside B (RB), rebaudioside D (RD), and steviol glycosides such as rebaudioside F (RF) , dextrin, water, etc.

Example 3: Sensory evaluation of diluted soju prepared using enzyme-treated stevia sweetener

Sensory evaluation of the diluted soju prepared in Example 2 was performed.

Meanwhile, the sensory evaluation characteristics of a single steviol glycoside according to the present invention are shown in Table 5 below. In this case, the degree of sweetness is based on sugar.

Steviol glycoside single component sweetness Sweetness/Bitterness Dulcoside A (DA) 40-60 The taste is weak and the bitterness is very strong. Stevioside (ST) 100-200 It has a sweet taste and a strong bitter aftertaste. Rebaudioside C (RC) 40-60 Weak taste and bitter taste Rebaudioside A (RA) 250-300 Strong taste, excellent sweetness in the aftertaste, and weak bitterness in the aftertaste

Specifically, for the T-1 to T-4 soju samples in Table 4 and commercially available soju products containing steviol glycosides, a 9-point scale method (1: very poor, 3: poor, 5: average, 7: good) , 9: very good) was evaluated for overall acceptability. It was conducted on 46 trained inspectors, and the overall preference, softness, cleanliness, and the taste of soju were evaluated. The sensory evaluation results are shown in Table 6 below. At this time, statistical analysis of the evaluation results verified a significant difference as a p-value using ANOVA analysis.

test sample overall symbolism Soft cleanliness taste of shochu T-1 6.0 6.3 6.2 6.0 T-2 5.8 5.6 5.0 5.7 T-3 6.1 6.5 6.4 6.1 T-4 5.9 5.3 5.1 5.4 Commercially available products containing steviol glycosides 4.3 4.4 4.7 4.8 p-Value ¹⁾ 0.009 0.017 0.023 0.049

¹⁾ Significant difference when N=46, p-Value≤0.05

As a result of sensory evaluation, as shown in Table 6 above, it was prepared by adding the enzyme-treated stevia sweetener according to the present invention prepared by glycosylation of a single steviol glycoside in a specific ratio compared to commercially available products containing steviol glycoside. It was found that all of the diluted soju samples T-1 to T-4 were remarkably excellent in overall acceptability. In addition, it showed a high degree of preference in terms of the overall preference as well as the detailed properties of soju, such as softness, cleanliness, and the quality of soju, and showed a statistically significant difference. In particular, in the case of the diluted soju samples of T-1 and T-3, it was confirmed that the overall acceptability and the detailed properties of the soju were superior compared to the diluted soju of T-2 and T-4.

Summarizing the above series of results, including dulcoside A (DA), stevioside (ST), rebaudioside C (RC), and rebaudioside A (RA) in a specific ratio as a single component of steviol glycoside When the enzyme-treated stevia sweetener obtained by glycosylation of the steviol glycoside composition (see Table 7) is added in an amount of 0.01 to 0.04% by weight based on the total weight when preparing diluted soju (see Table 8), It was found that in terms of detailed properties of shochu such as cleanliness and shochu-like properties, it was found to exhibit a significantly higher degree of preference compared to commercially available soju products containing steviol glycosides.

On the other hand, the enzyme-treated stevia sweetener with improved sweetness through HPLC analysis contains steviol glycoside G1 (G1) and steviol glycoside G2 (G2) newly generated by glycosylation, and Dulco It was confirmed that the side A had disappeared. That is, the enzyme-treated stevia sweetener finally obtained for application to the production of soju includes steviol glycoside G1 (G1), stevioside (ST), rebaudioside C (RC), rebaudioside A (RA), and It was confirmed that a single component of steviol glycoside G2 (G2) was included.

Steviol glycoside single component and its content included in the steviol glycoside composition Dulcoside A
(DA) Stevioside
(ST) Rebaudioside C
(RC) Rebaudioside A
(RA) content
(weight%) 0~3 45~55 1-5 35-45

Steviol glycoside single component and its content in diluted soju steviol
Glycoside G1
(G1) Stevio
side
(ST) rebaudio
side C
(RC) rebaudio
side A
(RA) steviol
Glycoside G2
(G2) content
(weight%) 0.0002~0.0010 0.0025~0.0060 0.0001~0.0005 0.0055~0.02 0.003-0.006

That is, from the above results, it can be seen that the enzyme-treated stevia sweetener obtained by enzymatically treating a steviol glycoside composition containing a single component of steviol glycoside in a specific ratio can be usefully used in the preparation of dilute soju with improved sensory evaluation. there was. In addition, it was found that the enzyme-treated stevia sweetener prepared with a single steviol glycoside in a specific ratio makes it possible to produce diluted soju of uniform quality without affecting the sweetening quality that may be different depending on the variety or crop of stevia leaves. .

Claims (10)

An enzyme comprising the step of glycosylating a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A Processed Stevia Sweetener Manufacturing Method.
According to claim 1,
The steviol glycoside composition is based on the total weight of the composition,
0 to 3% by weight of dulcoside A;
45 to 55% by weight of stevioside;
1 to 5% by weight of rebaudioside C; and
A method for producing an enzyme-treated stevia sweetener, comprising 35 to 45% by weight of rebaudioside A.
0 to 3 wt% of dulcoside A; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; and 35 to 45% by weight of rebaudioside A; An enzyme-treated stevia sweetener prepared by glycosylation of the included steviol glycoside composition.
4. The method of claim 3,
The enzyme-treated stevia sweetener includes steviol glycoside G1, stevioside, rebaudioside C, rebaudioside A, and steviol glycoside G2.
a) glycosylation of a steviol glycoside composition comprising dulcoside A, stevioside, rebaudioside C and rebaudioside A to obtain an enzyme-treated stevia sweetener;
b) adding the enzyme-treated stevia sweetener obtained in step a) to alcohol having an alcohol concentration of 50 vol%; and
c) purifying and filtering the alcohol of step b), followed by the steps of:
A method for producing diluted soju comprising a.
6. The method of claim 5,
The steviol glycoside composition of step a) contains 0 to 3% by weight of dulcoside A based on the total weight of the composition; 45 to 55% by weight of stevioside; 1 to 5% by weight of rebaudioside C; and 35 to 45% by weight of rebaudioside A.
6. The method of claim 5,
The method for preparing diluted soju, wherein the enzyme-treated stevia sweetener of step b) is added in an amount of 0.01 to 0.04% by weight based on the total weight of the soju.
6. The method of claim 5,
The number of steps c) is adjusted so that the alcohol concentration is 15 to 17 vol%, the dilution method of producing soju.
The method of claim 5, wherein the diluted soju, based on the total weight of soju, steviol glycoside G1 0.0002 to 0.0010% by weight; 0.0025 to 0.0060% by weight of stevioside; 0.0001 to 0.0005% by weight of rebaudioside C; 0.0055 to 0.02% by weight of rebaudioside A; and 0.003 to 0.006 wt% of steviol glycoside G2; The included, diluted soju manufacturing method.
A diluted soju comprising 0.01 to 0.04% by weight of the sweetener of claim 3.

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