Composition comprising glucosylated steviol glycosides

Abstract

The present application relates to a composition comprising glucosylated steviol glycosides and an organic acid, and to beverages comprising same.

Description

Composition comprising a glucose-transferred steviol glycoside

[0001] [Cross-reference with related applications]

[0002] The present application claims the benefit of priority based on Korean Patent Application No. 10-2025-0012718 filed on January 31, 2025, Korean Patent Application No. 10-2025-0119488 filed on August 26, 2025, and Korean Patent Application No. 10-2025-0212678 filed on December 29, 2025, and all contents disclosed in said Korean patent application documents are incorporated herein as part of the specification.

[0003] The present application relates to a composition comprising a glucose-transferred steviol glycoside and a beverage comprising the same.

[0004]

[0005] Consumer interest in maintaining health has steadily increased due to the need for a sustainable lifestyle. Above all, dietary habits are closely related to health, and recently, consumers have been paying close attention to improving their eating habits, such as reducing sodium or sugar intake. In particular, excessive sugar intake has become a more critical factor than ever before in dietary habits due to concerns regarding rapid spikes in blood sugar, increased blood triglycerides, and obesity.

[0006] In 2015, the WHO recommended lowering daily sugar intake to 25g due to concerns about diseases such as obesity, and accordingly, governments and companies in developed countries are taking action.

[0007] Currently, synthetic high-intensity sweeteners (such as aspartame and sucralose) are primarily used as alternative sweeteners to replace sugar. However, as concerns regarding the safety of synthetic sweeteners continue to be raised, there is a growing demand for healthier alternative sweeteners. Various natural sweetener materials used as alternatives often exhibit unsatisfactory sweetness quality, such as bitterness, off-flavors, or astringent tastes, leaving the challenge of resolving this issue unresolved.

[0008] Among these, stevia, a natural high-intensity sweetener considered to be highly popular among consumers, refers to a sweetener derived from the leaves of the plant *Stevia rebaudiana Bertoni*. Steviol glycosides, known as the main sweetening components, have a sweetness approximately 200 to 400 times that of sugar, and in particular, rebaudioside A, rebaudioside D, and rebaudioside M are mainly used as sweeteners in food and beverages. However, rebaudioside A has a distinctive bitter taste, and rebaudioside D and rebaudioside M have low solubility and high prices, which limits their application.

[0009] There is a need for measures to solve the problem of such steviol glycosides.

[0010]

[0011] [Prior Art Literature]

[0012] U.S. Patent Publication US 12053005 B2

[0013] Korean Registered Patent Publication KR 10-2421798

[0014] U.S. Patent Publication US 2024-0090551 A1

[0015]

[0016] The present application aims to improve the sensory properties of food by utilizing glucose-transferred stevia glycosides and organic acids, and to provide a composition and a beverage comprising said glucose-transferred stevia glycosides and organic acids.

[0017]

[0018] One aspect of the present application provides a composition comprising a glucose-transferred steviol glycoside and an organic acid, wherein the steviol glycoside comprises at least one selected from the group consisting of stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside O, rebaudioside N, rebaudioside I, rebaudioside M, rubusoside, steviolbioside, and dulcoside A, and the glucose-transferred steviol glycoside comprises one or more glucose atoms added to the glucose connected to the 19th carbon of the steviol glycoside via an α-1,6 bond.

[0019] Another aspect of the present application provides a beverage comprising the above composition.

[0020]

[0021] The present application will be described in detail below.

[0022]

[0023] The term "steviol glycoside" in this application refers to a natural sweetener having a form in which glucose, rhamnose, xylose, etc. are bonded to the 13th and 19th-OH groups of steviol of the following chemical formula 1.

[0024] [Chemical Formula 1]

[0025]

[0026] In Chemical Formula 1, hydrogen (H) may be bonded to R1, or one to three glucose molecules may be bonded via α- / β-bonds, and R2 may have one glucose molecule, xylose, or rhamnose molecule bonded via α- / β-bonds, and zero to two glucose molecules bonded via α- / β-bonds, but is not limited thereto.

[0027] The above-mentioned steviol glycoside may be at least one selected from the group consisting of stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside O, rebaudioside N, rebaudioside I, rebaudioside M, lubusoside, steviolbioside, and dulcoside A, but is not limited thereto.

[0028] The above steviol glycoside has the advantage of being low in calories compared to sugar and having a sweetness level about 200-300 times that of sugar, but it has the disadvantage of being accompanied by a unique astringent or bitter taste, so efforts have been made to improve the sweetness.

[0029] α- / β-glycosidic bonds are distinguished by the anomeric position and the relative stereochemistry (R- or S-type) of the stereocenter furthest from the 1st carbon of the monosaccharide. Generally, α-glycosidic bonds are formed when two carbons have the same stereochemistry, whereas β-glycosidic bonds occur when two carbons have different stereochemistry.

[0030] The term "glucose-transferred steviol glycoside" in this application may be a form in which one or more glucose molecules are added to the 19-OH position of a steviol glycoside via an α-bond, and more specifically, may be a form in which one or more glucose molecules are added to the glucose connected to the 19th carbon of the steviol glycoside via an α-1,6 bond, but is not limited thereto. Specifically, the glucose-transferred steviol glycoside may be in a form in which 1 to 11 molecules of glucose are added to the glucose connected to the 19th carbon of ribaudioside A via an α-1,6 bond, and more specifically, the number of added glucose molecules may be 1 to 11 molecules, 1 to 10 molecules, 1 to 9 molecules, 1 to 8 molecules, 1 to 7 molecules, 1 to 6 molecules, 1 to 5 molecules, 1 to 4 molecules, 2 to 11 molecules, 3 to 11 molecules, 4 to 11 molecules, 5 to 11 molecules, 6 to 11 molecules, 7 to 11 molecules, 8 to 11 molecules, 9 to 11 molecules, 10 to 11 molecules, or 11 molecules, but is not limited thereto.

[0031] The glucose-transferred steviol glycoside may be prepared by modifying it into a glucose-transferred steviol glycoside in the presence of a crude enzyme solution having glucose-transferring activity, a glucose-transferring enzyme, a microorganism expressing the same, or a culture of said microorganism, and a glucose donor, but is not limited to any method that a person skilled in the art can prepare by transferring glucose to a steviol glycoside. Specifically, the glucose-transferred steviol glycoside of the present application may be prepared by a step of reacting sugar with a steviol glycoside in the presence of a Lactobacillus mali microorganism or a culture thereof, and said Lactobacillus mali may include Lactobacillus maliKCCM13503P(LactobacillusmaliCJST242), DSM20444, ATCC 27054, or ATCC 27304, but is not limited thereto.

[0032] The sweetness of the glucose-transferred steviol glycoside of the present application may be about 60 times or more than that of sugar, specifically 66.8 times.

[0033]

[0034] One aspect of the present application provides a composition comprising the glucose-transferred stevia glycoside and an organic acid.

[0035] In various beverages, the content of the glucose-transferred steviol glycoside of the composition of the present application may be 0.001 to 1 part by weight based on 100 parts by weight of the beverage, and any one lower limit selected from the group consisting of 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, and 0.11 parts by weight, and 0.2, 0.3. It may be included in an amount within a range selected from the group consisting of any one upper limit selected from the group consisting of 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, and 1 part by weight, for example, the content of the glucose-transferred steviol glycoside may be 0.005 to 1 part by weight, 0.01 to 0.9 parts by weight, 0.02 to 0.9 parts by weight, 0.03 to 0.8 parts by weight, 0.04 to 0.8 parts by weight, 0.05 to 0.7 parts by weight, 0.06 to 0.6 parts by weight, 0.07 to 0.5 parts by weight, 0.08 to 0.4 parts by weight, 0.09 to 0.3 parts by weight, 0.1 to 0.2 parts by weight, or 0.11 to 0.2 parts by weight, based on 100 parts by weight of the beverage.

[0036] The above organic acid refers to a substance that is organic and acidic. This is a concept contrasted with inorganic acids, and organic acids are generally weak acids that do not dissolve well in water. Specifically, the above organic acid may be at least one selected from the group consisting of phosphoric acid, citric acid, malic acid, tartaric acid, lactic acid, formic acid, ascorbic acid, fumaric acid, gluconic acid, succinic acid, maleic acid, and adipic acid, but is not limited thereto.

[0037] In one embodiment, the organic acid may be phosphoric acid, citric acid, malic acid, or tartaric acid, but is not limited thereto.

[0038] In one embodiment, the organic acid may be phosphoric acid or citric acid, but is not limited thereto.

[0039] The composition of the present invention may be a beverage composition and may be a composition for use in making a beverage.

[0040] In various beverages, the content of the organic acid of the composition of the present application may be 0.0001 to 0.5 parts by weight based on 100 parts by weight of the beverage, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, and 0.001 parts by weight, and an upper limit selected from the group consisting of 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, and 0.5 parts by weight; for example, the content of the organic acid may be 0.0002 to 0.45 parts by weight, 0.0003 parts by weight, based on 100 parts by weight of the beverage. It may be up to 0.4 parts by weight, 0.0004 to 0.35 parts by weight, 0.0005 to 0.3 parts by weight, 0.0006 to 0.3 parts by weight, 0.0007 to 0.25 parts by weight, 0.0008 to 0.2 parts by weight, 0.0009 to 0.15 parts by weight, or 0.001 to 0.1 parts by weight.

[0041] When the above organic acid is phosphoric acid, the content of the above phosphoric acid may be 0.0005 to 0.45 parts by weight, 0.0007 to 0.4 parts by weight, 0.001 to 0.35 parts by weight, 0.001 to 0.3 parts by weight, 0.0013 to 0.3 parts by weight, 0.0015 to 0.2 parts by weight, 0.0016 to 0.17 parts by weight, 0.0017 to 0.15 parts by weight, 0.002 to 0.1 parts by weight, 0.0022 to 0.0396 parts by weight, 0.0022 to 0.025 parts by weight, 0.005 to 0.025 parts by weight, or 0.0019 to 0.0935 parts by weight, based on 100 parts by weight of the beverage.

[0042] When the above organic acid is citric acid, the content of the citric acid is, based on 100 parts by weight of the beverage, 0.0002 to 0.45 parts by weight, 0.0003 to 0.4 parts by weight, 0.0004 to 0.35 parts by weight, 0.0005 to 0.3 parts by weight, 0.0006 to 0.3 parts by weight, 0.0007 to 0.25 parts by weight, 0.0008 to 0.2 parts by weight, 0.0009 to 0.15 parts by weight, 0.001 to 0.3 parts by weight, 0.001 to 0.1 parts by weight, 0.0019 to 0.17 parts by weight, 0.005 to 0.025 parts by weight, 0.032 to 0.17 parts by weight, and 0.015 to 0.062 parts by weight. It may be 0.0019 to 0.3 parts by weight or 0.0019 to 0.09 parts by weight.

[0043]

[0044] In addition to the above, the composition of the present application may further include at least one additive selected from the group consisting of flavorings, nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectin, fruit pulp, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonating agents, and acidifiers. The nutrients may include vitamins or minerals. The content of such additives may be selected in the range of 0.001 to 90 parts by weight based on 100 parts by weight of the beverage of the present application.

[0045] The above composition may further include a fragrance.

[0046] The above flavoring may be a substance that imparts flavor to food. The above flavoring may include natural flavorings and artificial flavorings, and may be used without limitation as long as it is applicable to food. For example, the flavorings of the present application include cola flavor, cider flavor, beer flavor, ginger ale flavor, energy drink base flavor, gummy candy flavor, gum flavor, milk flavor, yogurt flavor, yogurt flavor, vanilla flavor, chocolate flavor, cocoa flavor, honey flavor, caramel flavor, cheese flavor, floral flavor, syrup flavor, grain flavor (barley, corn, brown rice, black rice, misugaru, oatmeal, walnut, etc.), nut flavor (almond, hazelnut, cashew, pistachio, etc.), fruit flavor (orange, lemon, lime, grapefruit, yuzu, calamansi, mandarin, pineapple, mango, banana, peach, apricot, plum, apple, pear, strawberry, raspberry, blueberry, blackberry, cherry, grape, cranberry, kiwi, melon, watermelon, coconut, passion fruit, guava, dragon fruit, lychee, quince, etc.), herb / plant flavor (basil, mint (peppermint, It may be a substance that produces spearmint), rosemary, thyme, lavender, chamomile, hibiscus, jasmine, lemongrass, honeybush, ginger, kudzu root, ginseng, green tea, black tea, mate, chlorella, aloe, etc., but is not limited thereto.

[0047] In the above composition, the content ratio of the glucose-transferred steviol glycoside to the organic acid may be 0.001 : 0.5 to 1 : 0.0001 or 0.001 : 0.5 to 0.5 : 0.0001, for example, 0.012 : 0.2 to 0.3 : 0.001, 0.08 : 0.1 to 0.03 : 0.0015 or 0.1197 : 0.001 to 0.1197 : 0.3.

[0048] In addition, the above composition may be used in the form of a liquid composition or a mixture of a liquid and a gel.

[0049] The above composition may be used for the manufacture of RTD (Ready to Drink) beverage products and may be a beverage composition, and the composition may also be used in the form of a semi-finished product or a syrup or powder during the process.

[0050] The composition of the present application can serve as a sweetener to impart sweetness to food. By including the glucose-transferred steviol glycoside and an organic acid, the bitterness of the glucose-transferred steviol glycoside and the sweetness preference can be improved, and the excellent sweetness quality can reduce off-flavors or off-odors. Furthermore, by utilizing the composition of the present application containing the glucose-transferred steviol glycoside and an organic acid, the problem of off-flavors or off-odors inherent in the steviol glycoside sweetener itself can be improved, and the sourness intensity of the organic acid can be lowered to improve taste quality. Consequently, food containing the composition of the present application can have excellent taste quality with high flavor preference and overall preference.

[0051]

[0052] In addition, the composition of the present application may be a sweetener composition for beverages.

[0053] In addition to glucose-transferred steviol glycosides, the above composition may further include a nutritional or non-nutritional sweetener. The sweetener may include at least one sweetener selected from the group consisting of non-nutritional sweeteners, allulose, fructose, glucose, mannose, arabinose, galactose, xylose, rhamnose, ribose, fucose, sucrose, maltose, lactose, maltitol, xylitol, erythritol, sorbitol, mannitol, palatnitol, maltotriitol, maltotetraitol, and D-tagatose. In various beverages, the content of the sweetener may be 1 to 90 parts by weight based on 100 parts by weight of the beverage, but is not limited thereto.

[0054] The composition of the present application may be added directly to a beverage as a sweetener or used in combination with other beverage ingredients, and may be used appropriately according to conventional methods. The composition of the present application may contain various flavoring agents or natural carbohydrates, etc., as additional ingredients.

[0055]

[0056] Another aspect of the present application provides a beverage comprising the above composition.

[0057] The content of the glucose-transferred steviol glycoside may be 0.001 to 1 part by weight based on 100 parts by weight of the beverage, and any one lower limit selected from the group consisting of 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, and 0.11 parts by weight, and 0.2, 0.3. It may be included in an amount within a range selected from the group consisting of any one upper limit selected from the group consisting of 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, and 1 part by weight, for example, the content of the glucose-transferred steviol glycoside may be 0.005 to 1 part by weight, 0.01 to 0.9 parts by weight, 0.02 to 0.9 parts by weight, 0.03 to 0.8 parts by weight, 0.04 to 0.8 parts by weight, 0.05 to 0.7 parts by weight, 0.06 to 0.6 parts by weight, 0.07 to 0.5 parts by weight, 0.08 to 0.4 parts by weight, 0.09 to 0.3 parts by weight, 0.1 to 0.2 parts by weight, or 0.11 to 0.2 parts by weight, based on 100 parts by weight of the beverage. The content of the organic acid may be 0.0001 to 0.5 parts by weight based on 100 parts by weight of the beverage, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, and 0.001 parts by weight, and an upper limit selected from the group consisting of 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, and 0.5 parts by weight; for example, the content of the organic acid may be 0.0002 to 0.45 based on 100 parts by weight of the beverage. parts by weight, 0.0003 to 0.4 parts by weight, 0.0004 to 0.35 parts by weight, 0.0005 to 0.3 parts by weight, 0.0006 to 0.3 parts by weight, 0.0007 to 0.It may be 25 parts by weight, 0.0008 to 0.2 parts by weight, 0.0009 to 0.15 parts by weight, or 0.001 to 0.1 parts by weight. In one embodiment, the organic acid may be phosphoric acid or citric acid.

[0058] When the above organic acid is phosphoric acid, the content of the above phosphoric acid may be 0.0005 to 0.45 parts by weight, 0.0007 to 0.4 parts by weight, 0.001 to 0.35 parts by weight, 0.001 to 0.3 parts by weight, 0.0013 to 0.3 parts by weight, 0.0015 to 0.2 parts by weight, 0.0016 to 0.17 parts by weight, 0.0017 to 0.15 parts by weight, 0.002 to 0.1 parts by weight, 0.0022 to 0.0396 parts by weight, 0.0022 to 0.025 parts by weight, 0.005 to 0.025 parts by weight, or 0.0019 to 0.0935 parts by weight, based on 100 parts by weight of the beverage.

[0059] When the above organic acid is citric acid, the content of the citric acid is, based on 100 parts by weight of the beverage, 0.0002 to 0.45 parts by weight, 0.0003 to 0.4 parts by weight, 0.0004 to 0.35 parts by weight, 0.0005 to 0.3 parts by weight, 0.0006 to 0.3 parts by weight, 0.0007 to 0.25 parts by weight, 0.0008 to 0.2 parts by weight, 0.0009 to 0.15 parts by weight, 0.001 to 0.3 parts by weight, 0.001 to 0.1 parts by weight, 0.0019 to 0.17 parts by weight, 0.005 to 0.025 parts by weight, 0.032 to 0.17 parts by weight, and 0.015 to 0.062 parts by weight. It may be 0.0019 to 0.3 parts by weight or 0.0019 to 0.09 parts by weight.

[0060] If the beverage of the present application contains the glucose-transferred steviol glycoside and / or organic acid in the above-mentioned content range, the sensory quality of the beverage can be improved, specifically, the sourness intensity of the beverage is lowered even when an acidifier such as an organic acid is added, the flavor preference of the beverage can be increased, and the overall preference can be increased.

[0061] The beverages of the present application include, but are not limited to, general beverages, health beverages, and medical (or patient) beverages.

[0062] Specifically, the beverage may be a carbonated or non-carbonated beverage and may include, for example, soft drinks, fountain drinks, frozen ready drinks (carbonated or non-carbonated), coffee drinks, tea drinks, brewed drinks other than coffee or tea, dairy drinks, flavored water, functionally enhanced beverages such as fortified water, juices such as fruit juice (including concentrated juices that can be diluted and drunk immediately), fruit juice-flavored drinks, sports drinks, smoothies, caffeine-containing energy drinks, or alcoholic products. In certain embodiments, the composition may be a carbonated beverage, for example, a carbonated cola-flavored beverage. The cola-flavored carbonated beverage is characterized by containing, in addition to the combination of rebaudioside disclosed herein, carbonated water, sweeteners, kola nut extract and / or other flavorings, caramel coloring, phosphoric acid, and optionally other ingredients.

[0063] Examples of the above juice sources may include plums, plums, figs, pineapples, peaches, bananas, apples, pears, guavas, apricots, watermelons, coconuts, olives, kiwis, quinces, sea buckthorn, passion fruit, rowan berries, pomegranates, persimmons, mangoes, rhubarb, papayas, lychees, lemons, oranges, limes, tangerines (citrus), mandarin oranges, tangelos, pomelos, grapefruits, Barbados cherries (acerola cherries), bearberries, blackberries, blueberries, boysenberries, cherries, choke cherries, cloudberries, cranberries, wild grapes, dates, dewberries, elderberries, grapes, gooseberries, huckleberries, loganberries, olaliberries, mulberries, raisins, plainsberries, prairie berries, raspberries, Saskatoon berries, salmonberries, sea buckthorn berries, sloe berries, strawberries, thimbleberries, thornberries, wineberries, lingonberries, etc., and at least specific Numerous additional and alternative juices suitable for use in the embodiments will be apparent to those skilled in the art, taking into account the advantages of this disclosure. The specific amount of juice useful for imparting flavor characteristics to a beverage product depends on the selected juice, the desired flavor impression, and the form of the juice ingredients. Taking into account the advantages of this specification, those skilled in the art will be able to easily determine the amount of any specific juice(s) used to achieve the desired flavor impression.

[0064] In one embodiment, the beverage may be cola.

[0065] In one embodiment, the beverage may be a lemon-lime carbonated drink or lemonade.

[0066] In one embodiment, the beverage may be a fruit and vegetable beverage, specifically an apple beverage or a citrus beverage.

[0067] The beverage may include at least one additive selected from the group consisting of flavorings, nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectin, fruit pulp, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents, and acidifiers. Based on 100 parts by weight of the beverage, the content of the additive may be 1 to 90 parts by weight.

[0068] Specifically, the above beverage may further include flavorings.

[0069] The above flavoring may be a substance that imparts flavor to food. The above flavoring may include natural flavorings and artificial flavorings, and may be used without limitation as long as it is applicable to food. For example, cola scent, soda scent, beer scent, ginger ale scent, energy drink base scent, gummy candy scent, gum scent, milk scent, yogurt scent, yogurt scent, vanilla scent, chocolate scent, cocoa scent, honey scent, caramel scent, cheese scent, floral scent, syrup scent, grain scent (barley, corn, brown rice, black rice, misugaru, oatmeal, walnut, etc.), nut scent (almond, hazelnut, cashew, pistachio, etc.), fruit scent (orange, lemon, lime, citrus, grapefruit, yuzu, calamansi, mandarin, pineapple, mango, banana, peach, apricot, plum, apple, pear, strawberry, raspberry, blueberry, blackberry, cherry, grape, cranberry, kiwi, melon, watermelon, coconut, passion fruit, guava, dragon fruit, lychee, quince, etc.), herb / plant scent (basil, mint (peppermint, spearmint), It may be a substance that produces or causes fruit and vegetable scents, such as rosemary, thyme, lavender, chamomile, hibiscus, jasmine, lemongrass, honeybush, ginger, kudzu root, ginseng, green tea, black tea, mate, chlorella, aloe, etc.), but is not limited thereto.

[0070] The pH of the beverage of the present application may be from pH 2.0 to 5.0, and specifically, the pH of the beverage may be a range selected from the group consisting of any one lower limit selected from the group consisting of pH 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, and 2.7, and any one upper limit selected from the group consisting of pH 3.5, 4.0, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, and 5.0, for example, the pH of the beverage may be pH 2.1 to 4.9, pH 2.2 to 4.8, pH 2.3 to 4.7, pH 2.4 to 4.6, pH 2.5 to 4.5, pH 2.6 to 4.4, pH 2.8 to 4.0, or pH 2.7 to 3.5. Since the composition of the application contains an organic acid, a beverage food containing said composition may exhibit acidity; in this case, if the pH of said beverage is within the above numerical range, the sourness intensity of the beverage may be reduced, and flavor preference and overall preference may be improved.

[0071] When the organic acid is phosphoric acid, the pH of the beverage may be from pH 2.0 to 5.0, and specifically, the pH of the beverage may be a range selected from the group consisting of a lower limit selected from the group consisting of pH 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, and 2.7, and an upper limit selected from the group consisting of pH 3.5, 3.7, 3.9, 4.0, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, and 5.0, for example, when the organic acid is phosphoric acid, the pH of the beverage may be from pH 2.1 to 4.9, pH 2.2 to 4.8, pH 2.3 to 4.7, pH 2.4 to 4.6, pH 2.5 to 4.5, pH 2.6 to 3.9, The pH may be 2.64 to 3.83 or 2.7 to 3.5, but is not limited thereto.

[0072] When the organic acid is citric acid, the pH of the beverage may be from pH 2.0 to 5.0, and specifically, the pH of the beverage may be a range selected from the group consisting of a lower limit selected from the group consisting of pH 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, and 2.8, and an upper limit selected from the group consisting of pH 3.3, 3.5, 3.7, 4.0, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, and 5.0; for example, when the organic acid is citric acid, the pH of the beverage may be pH 2.1 to 4.9, pH 2.2 to 4.8, pH 2.3 to 4.7, pH 2.4 to 4.6, pH 2.5 to 4.5, pH 2.5 It may be up to 4.4, pH 2.51 to 4.35, pH 2.8 to 4.0, or pH 2.9 to 3.3, but is not limited thereto.

[0073]

[0074] The glucose-transferred steviol glycoside of the present application has improved bitterness and sweetness palatability, and has excellent sweetness quality with minimal off-flavors or odors. By utilizing a composition containing such a glucose-transferred steviol glycoside and an organic acid, the off-flavors or odors of the steviol glycoside sweetener can be improved, and the sourness intensity of the organic acid can be lowered to improve taste quality; thus, food containing the composition of the present application has excellent taste quality with high flavor palatability and overall palatability.

[0075] In addition, it was confirmed that the glucose-transferred steviol glycoside used in this application has high stability when combined with organic acids. Through the combination of this glucose-transferred steviol glycoside with various organic acids, bitterness, astringency, and sourness can be improved, and excellent sweetness can be achieved.

[0076]

[0077] Figure 1 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside A by the Lactobacillus mali CJST242 strain.

[0078] Figure 2 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside C by the Lactobacillus mali CJST242 strain.

[0079] Figure 3 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside F by the Lactobacillus mali CJST242 strain.

[0080] Figure 4 is an HPLC chromatogram of the result of the glycosylation reaction of stevioside by the Lactobacillus mali CJST242 strain.

[0081] Figure 5 is an HPLC chromatogram of the result of the glycosylation reaction of dulcosides by the Lactobacillus mali CJST242 strain.

[0082] Figure 6 is an HPLC chromatogram of the glycosylation reaction results of rubusoside by the Lactobacillus mali CJST242 strain.

[0083] Figure 7 shows the chemical structure of glucose-transferred ribaudioside A in which 1 to 4 glucose molecules are added to the glucose connected to the 19th carbon of ribaudioside A through an α-(1,6) bond.

[0084]

[0085] The present application will be explained in more detail below through examples.

[0086] However, these examples are provided only to aid in understanding the present application and do not limit the scope of the present application to these examples in any way.

[0087]

[0088] Preparation Example 1: Preparation of glucose-transferred steviol glycosides

[0089] The preparation of glucose-transferred steviol glycosides was carried out according to the method described in Korean Patent Publication No. 10-2021-0114899.

[0090] Lactobacillus mali (Lactobacillus mali KCCM13503P (Lactobacillus mali CJST242)) microorganisms were cultured at 30°C for 24 hours in a nutrient medium containing sugar (Baekseol white sugar with a purity of 99% or higher) as a carbon source, yeast extract and corn steep liquor as nitrogen sources, and amino acids. The above Lactobacillus mali CJST242 strain is a strain deposited with the Korean Culture Center of Microorganisms (KCCM) on August 5, 2024, under accession number KCCM 13503P. The culture medium was centrifuged at 8000 rpm for 10 minutes to separate the cells and the supernatant, and only the supernatant was collected. The crude enzyme solution was reacted with sugar to confirm the sugar hydrolysis activity, and the crude enzyme solution was used to prepare the following glucose-transferred steviol glycosides.

[0091] After dissolving the steviol glycoside and sugar in a 0.05 M acetate buffer solution, the crude enzyme solution of the Lactobacillus mali strain prepared in Preparation Example 1-1 was added, and the mixture was reacted at 40°C for 24 hours. After the reaction, the mixture was inactivated at 100°C, and the production of glucose-transferred steviol glycosides was confirmed by HPLC. The steviol glycosides used were stevioside, rubusoside, dulcoside A, and ribaudioside A / C / F, and HPLC was used to confirm whether glucose-transferred stevioside, glucose-transferred rubusoside, glucose-transferred dulcoside A, and glucose-transferred ribaudioside A / C / F were produced (refer to Figures 1 to 6). Glucose-transferred ribaudioside A, in which 1 to 11 glucose molecules are added to the glucose connected to the 19th carbon of the above ribaudioside A via an α-(1,6) bond, was used in the experiment as the glucose-transferred steviol glycoside of the present application.

[0092] NMR analysis was performed to confirm the structure of glucose-transferred ribaudioside A, in which 1 to 4 glucose molecules are added to the glucose molecule connected to the 19th carbon of the ribaudioside A via an α-(1,6) bond.

[0093] Specifically, the reaction solution in which the glycosylation reaction was completed was inactivated at 100°C to inactivate the enzyme, and impurities were removed using a 0.45 μm filter. Subsequently, the reaction solution from which impurities had been removed was loaded onto a column packed with an adsorption resin (LXS-869, Sunresin), and after elution using 70% ethanol, the eluent was passed through an anion exchange resin (LXS-865, Sunresin) and vacuum concentrated. To separate each substance in which 1 to 4 glucose molecules were transferred to ribaudioside A from the concentrated eluent, the eluent was loaded onto a chromatography column. Reb A-G1, Reb A-G2, Reb A-G3, and Reb A-G4 were fractionated using a column packed with C18 resin (ODS-AQ-HG, YMC) and an FPLC system (AKTA avant), and then evaporated in a 105°C dry oven to prepare solid samples of ribaudioside A with 1 to 4 glucose transfers. Approximately 10 mg of each test sample was dissolved in 600 μL of D2O, filtered, and transferred to a High Field NMR sample tube. To analyze the binding structure of each isolated glucose-transferred ribaudioside A 1 H / 13 It was confirmed by C NMR, homonuclear correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), heteronuclear single-quantum coherence (HSQC), and heteronuclear multiple-bond correlation (HMBC), and 2D rotating frame Nuclear Overhauser Effect Spectroscopy (ROESY).

[0094] The results are listed in Tables 1 to 4 below.

[0095] As a result of confirming the structures of substances in which one to four glucose units are transferred to ribaudioside A (RebA), the substance in which one glucose unit is transferred was identified as RebA-G1, defined as (13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy] entkaur-16-en-19-oic acid 6-O-α-D-glucopyranosyl-β-D-glucopyranosyl ester), which was identified as having a structure in which α-D-glucopyranose is α-(1,6) bonded to the 6th position of glucopyranose bonded to the 19th carbon based on the structure of ribaudioside A (RebA), a stevioside derivative. (Hereinafter, the α-(1,6) bonded α-D-glucopyranose is referred to as "sugar E").

[0096] In addition, the substance with two glucose transfers was identified as RebA-G2a, (13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy] entkaur-16-en-19-oic acid 6-O-α-D-glucopyranosyl-(1→6)-O-α-D-glucopyranosyl-β-D-glucopyranosyl ester), which has a structure in which α-D-glucopyranose is α-(1,6) linked to the 6th position of sugar E in RebA-G1. (Hereafter, the α-D-glucopyranose α-(1,6) linked to sugar E is referred to as "sugar F").

[0097] In addition, the substance with three transferred glucose units was identified as RebA-G3a, (13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy] entkaur-16-en-19-oic acid 6-O-α-D-glucopyranosyl-(1→6)-O-α-D-glucopyranosyl-(1→6)-O-α-D-glucopyranosyl-β-D-glucopyranosyl ester), which is a structure in which α-D-glucopyranose is α-(1,6) linked to the 6th position of sugar F in the RebA-G2a structure.

[0098] In addition, the substance with four transferred glucose units was identified as RebA-G4a, which has the structure of 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy] ent-kaur-16-en-19-oic acid 6-O-α-D-glucopyranosyl-(1→6)-O-α-D-glucopyranosyl-(1→6)-O-α-D-glucopyranosyl-(1→6)-O-α-D-glucopyranosyl-β-D-glucopyranosyl ester.

[0099] No.dH mult. (J in Hz)dC multHMBC CorrelationCOSY corr.Key ROESY corr.10.80 br m1.81 br m40.0CH239.21.39, 1.75, 1.810.80, 1.39, 1.750.95, 1.1221.39 br m1.75 br m18.6CH20.80, 1.04, 1.75, 1.810.80, 1.04, 1.39, 2.061.8131.04 br t (13.0)2.06 br m ovlp37.3CH227.9, 178.91.39, 1.75, 2.061.04, 1.39w, 2.06w1.12, 1.181.18443.8C51.12 br d (11.0)56.6CH15.1, 21.4, 27.9, 39.2, 43.8, 178.91.76, 1.810.80, 0.95, 1.04, 1.18, 1.39 (or 1.38), 1.8161.76 br m1.81 br m21.4CH21.12, 1.381.123.33, 3.4371.38 br m1.51 br m40.7CH253.1, 56.61.51, 1.761.38,1.183.33, 3.43841.9C90.95 br d (8.0)53.1CH15.1, 20.1, 36.4, 39.2, 40.7, 41.9, 44.0, 46.9w1.560.80, 1.12, 1.38, 1.51, 1.56, 2.001039.2C111.56 br m1.77 br m20.1CH20.95, 1.77, 1.901.56, 1.900.95, 1.902.10121.48 br m1.90 br m36.4CH287.4, 153.81.901.48, 1.56, 1.770.95, 2.10, 3.20, 3.33(or 3.34), 3.43(or 3.44), 3.660.83, 1.44, 2.10w 4.701387.4C141.44 br m 2.10 br m44.0CH236.4, 41.9, 53.1, 87.441.9, 46.9, 87.4w2.101.441.90, 3.33, 3.43, 4.700.83, 1.48, 3.20, 4.70152.00 br d (17.5)2.13 br d (17.5)46.9CH244.053.1, 87.4w, 153.82.13, 4.87, 5.062.00, 4.87, 5.060.95, 3.33(or 3.34), 3.43(or 3.44), 3.66, 4.851.44, 3.33(or 3.34), 3.43(or 3.44), 3.66, 4.8516153.4C174.87 br s5.06 br s104.5CH246.9, 87.446.9, 87.42.00, 2.132.00, 2.132.00, 2.13, 3.47, 3.66, 4.701.48w, 3.21, 3.33, 3.59, 3.66, 3.81, 4.70, 4.80181.18 s27.9CH318.6w, 37.3, 43.8, 56.6, 178.91.04, 1.12, 1.81, 2.0619178.9C200.83 s15.1CH339.2, 40.0, 53.1, 56.61.12, 1.75, 1.81, 1.90, 2.10, 3.203.33, 3.43, 3.59, 3.661'5.40 br d (8.0)94.1CH75.3, 76.3, 178.93.421.81w, 2.06w, 3.42, 3.49, 3.682'3.42 br m71.9CH76.3, 94.13.30~3.49 / 3.64~3.843'3.49 br m*76.3aCH69.0, 71.95.404'3.48 br m*69.0CH65.2, 75.35'3.68e br m*75.3bCH5.406'3.67e br m*3.89 br dd (11.0, 4.0)65.2CH297.897.83.893.673.49, 4.851''4.70 ovlp solv.95.9CH75.2, 78.7, 85.0, 87.43.661.44, 1.90, 2.10, 3.33, 3.822''3.66 br m78.7CH85.0, 95.9, 102.14.703''3.82 br m85.0CH68.6, 78.7, 95.9w, 102.23.433.33, 4.704''3.43 br m68.6CH60.8, 85.03.30~3.49 / 3.64~3.845''3.33f br m75.2bCH60.83.43, 3.64, 3.79w3.82, 4.706''3.64g br m*3.79 br m*60.8cCH23.33, 3.793.641'''4.80 br d (8.0)102.1CH76.4, 78.73.203.32, 3.39, 3.66, 5.062'''3.20 br t (8.0)74.2CH76.1, 102.13.39, 4.803'''3.39 br m76.1dCH70.2, 74.2, 102.1w3.20, 3.214.804'''3.21 br t (8.5)70.2CH61.5, 76.43.32, 3.393.59, 3.815'''3.32f br m76.4aCH3.21, 3.594.806'''3.59 br m3.81 br m*61.4CH276.43.32, 3.813.591''''4.72 br d (8.5)102.2CH73.4, 75.8, 85.03.303.33, 3.44, 3.822''''3.30 br t (8.5)73.4CH75.8, 102.23.44, 4.723''''3.44h br m75.8dCH69.5, 102.2w3.30~3.49 / 3.64~3.844''''3.34f br m69.5CH5''''3.33h br m75.7dCH3.30~3.49 / 3.64~3.846''''3.64 br m*3.84 br m60.6cCH269.53.843.43, 3.641'''''4.85 br d (4.0)97.8CH65.2, 71.7, 73.13.463.46, 3.67, 3.892'''''3.46 br m71.5CH97.8,3.65, 4.853'''''3.65g br m*73.1CH69.3, 71.53.36, 3.464'''''3.36 br m69.3CH60.4, 71.7, 73.13.61, 3.655'''''3.61 br m71.7CH3.36, 3.68, 3.816'''''3.68e br m3.81 br m*60.4cCH23.61.

[0100] w Weak signals.

[0101] * The chemical shift of each signal may not be exact value due to signal overlapping

[0102] a,b,c,d,e,f,g,hThe assignment of signals at same alphabet column may be exchangeable.

[0103]

[0104] No.dH mult. (J in Hz)dC multHMBC CorrelationCOSY corr.Key ROESY corr.10.80 br m1.82 br m39.9CH21.39, 1.73, 1.820.800.96, 1.05, 1.131.3921.39 br m1.73 br m18.6CH20.80, 1.05, 1.730.80, 1.391.052.05w31.05 br m2.05 br m37.2CH2179.11.39, 1.73, 2.051.050.80, 1.18, 1.39w1.18, 1.39443.8C51.13 br d (11.5)56.5CH15.1, 21.4, 39.1, 43.8, 179.11.76, 1.800.80, 0.96, 1.05, 1.18, 1.39, 1.8061.76 br m1.80 br m21.4CH21.13, 1.39, 1.801.13, 1.39, 1.761.491.1871.39 br m1.49 br m40.7CH256.51.491.390.96, 1.13, 1.800.96, 1.76841.9C90.96 br d (7.5)53.0CH15.1, 20.0, 36.4, 39.1, 39.9, 41.9, 44.1, 46.81.550.80, 1.13, 1.39, 1.48, 1.55, 2.011039.1C111.55 br m1.75 br m20.0CH241.9, 87.50.96, 1.75, 1.891.55, 1.890.96, 1.892.09121.48 br m1.89 br m36.4CH287.51.55, 1.891.48, 1.750.96, 2.090.83, 2.09, 4.701387.5C141.42 br m 2.09 br m44.1CH236.4, 41.9, 53.0, 87.546.8, 87.5w2.091.421.89, 4.700.83, 1.48, 1.75, 1.89, 3.20, 4.70152.01 br d (17.5)2.12 br d (17.5)46.8CH2153.52.12, 4.87, 5.042.01, 4.87, 5.040.96, 3.35, 3.44, 3.64, 4.871.42, 1.48, 3.35, 3.44, 3.64, 4.8716153.5C174.87 br s 5.05 br s104.3CH246.8, 87.546.8, 87.52.01, 2.122.01, 2.122.01, 2.101.48w, 3.33(or 3.32), 3.64, 3.80, 4.70, 4.80181.18 s27.9CH337.2, 43.8, 56.5, 179.11.05, 1.13, 1.80, 2.0519179.1C200.83 s15.1CH339.1, 39.9, 53.0, 56.51.75, 1.82, 1.89, 2.09, 3.20, 3.33, 3.43, 3.611'5.41 br d (8.5)94.1CH75.3, 76.3, 179.13.421.80w, 2.05w, 3.49, 3.702'3.42 br m71.9CH76.3, 94.15.413'3.49 br m*76.3aCH69.0, 75.35.414'3.48 br m*69.0CH76.35'3.70 br m75.3bCH5.416'3.70 br m3.89 br m65.3CH269.0, 97.73.883.683.48, 4.861''4.70 ovlp D2O signal95.9CH75.2, 78.7, 87.53.661.42, 1.89, 2.09, 3.33, 3.822''3.66 br m78.7CH84.9, 95.9, 102.13.82, 4.704.803''3.82 br m84.9CH68.6, 78.7, 102.23.43, 3.663.33, 4.704''3.43 br m68.6CH60.8 75.2, 78.7, 84.93.33, 3.825''3.33 br m75.2bCH3.43, 3.643.82, 4.706''3.64 br m*3.80 br m*60.8cCH23.33, 3.803.641'''4.80 br d (8.0)102.1CH75.8, 76.4, 78.73.203.33, 3.38, 3.662'''3.20 t (8.0)74.2CH75.8, 102.13.38, 4.803'''3.38 br m75.8CH70.2, 74.23.20, 3.224.804'''3.22 t (8.5)70.2CH61.3, 75.8, 76.43.33, 3.383.61, 3.815'''3.33 br m76.4aCH61.3, 70.23.22, 3.614.806'''3.61 br m*3.81 br m*61.3CH23.813.33, 3.611''''4.71 ovlp D2O signal102.2CH73.4, 75.8, 84.93.303.33, 3.44, 3.822''''3.30 br t (8.5)73.4CH75.8, 102.23.44, 4.713''''3.44 br m75.8dCH102.23.304''''3.35 br m*69.5eCH5''''3.33 br m*76.1dCH60.63.656''''3.65 br m*3.84 br m*60.6cCH23.33, 3.823.651'''''4.86 br s97.7CH65.3, 70.2, 71.4(or 71.5), 73.4(or 73.1)3.483.48, 3.70, 3.892'''''3.48 br m71.4fCH73.44.863'''''3.64 br m*73.4gCH69.3(or 69.4), 71.5(or 71.4)4'''''3.43 br m*69.4eCH65.33.805'''''3.80 br m*70.2CH3.436'''''3.63 br m*3.89 br m65.3CH297.797.71''''''4.87 br s97.7CH65.3, 71.8, 71.4(or 71.5), 73.4(or 73.1)3.473.47, 3.63, 3.892''''''3.47 br m71.5fCH4.873''''''3.64 br m*73.1gCH69.3(or 69.4), 71.5(or 71.4)4''''''3.36 br m*69.3eCH60.43.63(or 3.64)5''''''3.63 br m*71.8CH6''''''3.68 br m3.76 br m60.4CH2.

[0105] No.dH mult. (J in Hz)dC multHMBC CorrelationCOSY corr.Key ROESY corr.10.81 br m1.81 br m39.9CH239.1, 52.91.39, 1.72, 1.810.810.96, 1.14, 1.810.81, 0.82, 1.3921.39 br m1.72 br m18.5CH20.81, 1.06, 1.720.81, 1.06, 1.39, 1.81, 2.040.96, 1.06, 1.812.04w31.06 br m2.04 br m37.1CH2179.11.39, 1.72, 2.041.06, 1.39, 1.720.81, 1.14, 1.39w1.18, 1.39, 1.72443.8C51.14 br d (11.5)56.4CH15.1, 21.4, 27.9, 39.1, 43.8, 179.11.75, 1.800.81, 0.96, 1.06, 1.39, 1.8061.75 br m1.80 br m21.4CH21.14, 1.391.14, 1.391.481.1471.39 br m1.48 br m40.6CH21.48, 1.75, 1.801.390.96, 1.140.96, 1.75841.9C90.96 br d (8.0)52.9CH15.1, 20.0, 36.4, 39.1, 39.9, 40.6, 41.9, 44.1, 46.81.560.81, 1.14, 1.39, 1.48, 1.56, 2.021039.1C111.56 br m1.77 br m20.0CH239.139.1, 41.9, 87.60.96, 1.77, 1.901.56, 1.900.96, 1.902.09121.48 br m1.90 br m36.4CH287.6, 153.61.77, 1.901.48, 1.56, 1.770.96, 2.090.82, 1.42, 4.711387.6C141.42 br m 2.09 br m44.1CH236.4, 41.9, 52.9, 87.646.8, 87.6, 153.62.091.421.90, 4.710.82, 1.48, 1.77, 1.90, 4.71152.02 br d (17.0)2.12 br d (17.0)46.8CH244.1, 52.9153.62.12, 4.87, 5.042.02, 4.87, 5.040.96, 1.42, 1.56, 3.33-3.35, 3.64(or 3.65), 4.871.42, 1.48, 3.33- 3.35, 3.64(or 3.65), 4.8716153.6C174.87 br s 5.04 br s104.3CH246.8, 87.646.8, 87.62.02, 2.122.02, 2.122.02, 2.12, 3.821.48w, 3.33-3.35, 3.64, 3.79, 4.71, 4.80181.18 s27.9CH337.1, 43.8, 56.4, 179.11.06, 1.80, 2.0419179.1C200.82 s15.1CH339.1, 39.9, 52.9, 56.41.14, 1.72, 1.75, 1.81, 1.90, 2.09, 3.20, 3.33, 3.591'5.41 d (8.0)94.1CH75.3, 76.2, 179.13.423.49, 3.702'3.42 br m71.9CH76.2, 94.15.413'3.49 br m*76.2aCH69.05.414'3.49 br m*69.0CH76.25'3.70 br m*75.3bCH3.495.416'3.70 br m3.91 br m65.4cCH275.297.73.903.714.874.871''4.71 ovlp D2O signal95.9CH75.8, 78.7w, 84.9, 87.63.661.42, 1.90, 2.09, 3.33, 3.822''3.66 br m78.7CH84.9, 95.9, 102.14.713''3.82 br m84.9CH68.6, 78.7, 102.23.433.33, 4.714''3.43 br m*68.6CH60.8, 75.2, 78.7, 84.93.33-3.35, 3.825''3.33-3.35 br m*75.2bCH3.43, 3.644.716''3.64 br m,*3.79 br m*60.8dCH23.33-3.35, 3.793.641'''4.80 d (8.0)102.1CH75.8, 76.4, 78.73.203.33, 3.38, 3.66, 5.042'''3.20 t (8.0)74.1CH75.8, 102.13.38, 4.803'''3.38 br m75.8aCH70.2, 74.13.20, 3.224.804'''3.22 t (8.5)70.2eCH61.3, 76.43.33, 3.383.59, 3.815'''3.33 br m76.4CH3.22, 3.594.806'''3.59 br m3.81 br m61.3CH23.33, 3.813.593.221''''4.72 ovlp D2O signal102.2CH73.4, 75.8, 84.93.303.34, 3.44, 3.822''''3.30 br t (8.5)73.4CH75.8, 102.23.44, 4.723''''3.44 br m*75.8aCH102.23.304.724''''3.33-3.35 br m*69.5dCH5''''3.34 br m*76.1aCH60.63.656''''3.65 br m*3.82 br m*60.6dCH23.34, 3.823.651'''''4.87 br m*97.7gCH65.4, 70.1 or 70.2, 71.4 or 71.5, 73.1 or 73.43.483.48, 3.70, 3.912'''''3.48 br m*71.4fCH97.7(or 97.8)4.873'''''3.65 br m*73.4CH4'''''3.45 br m*69.5dCH5'''''3.80 br m*70.1eCH6'''''3.63 br m*3.90 br m65.4cCH297.7(or 97.8)70.1(or 70.2), 97.81''''''4.87 br m*97.7gCH65.4, 70.1 or 70.2, 71.4 or 71.5, 73.1 or 73.43.483.48, 3.63, 3.902''''''3.48 br m*71.4fCH97.7(or 97.8)4.873''''''3.66 br m*73.4CH4''''''3.44 br m*69.4dCH3.805''''''3.80 br m70.2eCH23.436''''''3.63 br m*3.90 br m65.3cCH297.8(or 97.7)70.1(or 70.2), 97.81'''''''4.87 br m*97.8g65.3, 70.1 or 70.2, 71.4 or 71.5, 73.1 or 73.43.493.49, 3.63, 3.902'''''''3.49 br m*71.5fCH97.8(or 97.7)4.873'''''''3.65 br m*73.1CH4'''''''3.36 br m*69.4dCH3.64-3.655'''''''3.64 br m*71.8CH6'''''''3.70 br m3.75 br m60.4CH2.

[0106] No.dH mult. (J in Hz)dC multHMBC CorrelationCOSY corr.Key ROESY corr.10.82 br m1.81 br m39.7CH21.38, 1.810.820.97, 1.16, 1.811.17, 1.38,21.38 br m1.70 br m18.5CH20.82, 1.07w, 1.700.82, 1.38, 2.031.81, 2.030.82, 2.0331.07 br td (13.5, 4.0)2.03 br d (13.5)37.1CH2179.21.38, 1.70, 2.031.03, 1.38, 1.701.17, 1.381.16, 1.38, 1.70443.8C51.16 br d (12.5)56.3CH15.1, 21.3, 37.1, 39.1, 40.6, 43.8, 179.21.74, 1.810.82, 0.97, 1.07, 1.4161.74 br m1.81 br m21.3CH21.16, 1.410.82, 1.47, 3.45, 3.64, 3.803.45, 3.64, 3.8071.41 br m1.47 br m40.6CH21.47, 1.74, 1.811.41, 1.740.97, 1.16841.8C90.97 br d (8.0)52.7CH15.1, 19.9, 39.1, 41.8, 44.11.560.82, 1.16, 1.38w, 1.48w, 1.56, 2.031039.1C111.56 br m1.76 br m19.9CH239.141.8, 87.60.97, 1.76, 1.891.56, 1.890.97, 1.891.48121.48 br m1.89 br m36.4CH287.61.56, 1.891.48, 1.761.76, 2.100.82, 2.10, 4.721387.6C141.42 br m 2.10 br d (13.0)44.1CH287.646.72.101.424.720.82, 0.97, 1.76, 1.89, 4.72w152.03 br d (15.5)2.11 br d (15.5)46.7CH2153.7153.72.10, 4.87, 5.032.02, 4.87, 5.030.97, 1.56, 3.32- 3.35, 3.64(or 3.65), 4.871.42, 1.48, 3.32- 3.35, 3.64(or 3.65), 4.8716153.7C174.87 br s 5.03 br s104.2CH246.7, 87.646.7, 87.62.03, 2.112.03, 2.112.03, 2.113.33, 3.59, 3.81, 4.71, 4.80181.17 s27.8CH337.1, 43.8, 56.3, 179.21.07, 1.81, 2.03w19179.2C200.82 s15.1CH339.1, 39.7, 52.7, 56.31.16, 1.70, 1.76 1.81, 1.89, 2.10, 3.22w1'5.41 d (8.0)94.1CH75.3, 76.3, 179.23.423.49, 3.702'3.42 br m71.9CH76.2, 94.15.413'3.49 br m*76.2aCH69.05.414'3.49 br m*69.0CH76.23.705'3.70 br m*75.3bCH69.03.495.416'3.70 br m3.90 br m65.4cCH275.397.73.903.704.884.881''4.72 ovlp D2O signal95.9CH78.7, 84.9, 87.73.661.42, 1.89, 2.10, 3.33, 3.822''3.66 br m78.7CH84.9, 95.9, 102.03.82, 4.723''3.82 br m84.9CH68.6, 78.7, 102.23.43, 3.664.724''3.43 br m*68.6CH60.8, 75.2, 78.7, 84.93.32-3.35, 3.825''3.32-3.35 br m*75.2bCH3.43, 3.644.726''3.64 br m*3.79 br m*60.8dCH23.32-3.35, 3.793.641'''4.80 d (8.0)102.0CH75.8, 76.3 78.73.203.33, 3.38, 3.662'''3.20 t (8.0)74.1CH75.8, 102.13.38, 4.803'''3.38 br m75.8aCH70.2, 74.13.20, 3.224.804'''3.22 t (8.5)70.2eCH61.3, 75.8, 76.43.33, 3.383.59, 3.815'''3.33 br m76.4CH61.33.22, 3.594.806'''3.59 br m3.81 br m61.3CH276.43.33, 3.813.33, 3.591''''4.71 ovlp D2O signal102.2CH73.4, 75.8, 84.93.303.33, 3.44, 3.822''''3.30 br t (8.5)73.4CH75.8, 102.23.44, 4.713''''3.44 br m*75.8aCH3.304.714''''3.35 br m*69.4dCH75.8, 76.15''''3.32-3.33 br m*76.1aCH60.63.656''''3.65 br m*3.82 br m*60.6dCH269.43.32-3.33, 3.823.651'''''4.86-4.90 br m*97.7gCH65.4, 70.0 or 70.2, 71.4 or 71.5, 73.1 or 73.43.44-3.503.44-3.50, 3.902'''''3.44-3.50 br m*71.4fCH4.86-4.903'''''3.61-3.67 br m*73.4CH71.44'''''3.45 br m*69.5dCH65.45'''''3.80 br m70.0eCH6'''''3.62-3.65 br m*3.88-3.92 br m*65.4cCH297.870.0, 97.81''''''4.86-4.90 br m*97.8gCH65.4, 70.0 or 70.2, 71.4 or 71.5, 73.1 or 73.43.44-3.503.44-3.50, 3.62- 3.65, 3.88-3.922''''''3.44-3.50 br m*71.4fCH4.86-4.903''''''3.66 br m*73.4CH71.44''''''3.44 br m*69.4dCH65.43.805''''''3.80 br m*70.2eCH23.446''''''3.62-3.65 br m*3.88-3.92 br m*65.4cCH297.870.2, 97.81'''''''4.86-4.90 br m*97.8gCH65.4, 70.0 or 70.2, 71.4 or 71.5, 73.1 or 73.43.44-3.503.44-3.50, 3.62- 3.65, 3.88-3.922'''''''3.44-3.50 br m*71.4fCH4.86-4.903'''''''3.66 br m*73.4CH69.4(or 69.5), 71.4(or 71.5)4'''''''3.44 br m*69.4dCH65.33.805'''''''3.80 br m70.2eCH23.446'''''''3.62-3.65 br m*3.88-3.92 br m*65.3cCH297.670.2, 97.61''''''''4.86-4.90 br m*97.6g65.3, 70.0 or 70.2, 71.4 or 71.5, 73.1 or 73.43.44-3.503.44-3.50, 3.62- 3.65, 3.88-3.922''''''''3.44-3.50 br m*71.5fCH4.86-4.903''''''''3.65 br m*73.1CH69.5(or 69.4), 71.5(or 71.4)4''''''''3.36 br m*69.5dCH71.83.64-3.655''''''''3.64 br m*71.8CH6''''''''3.69 br m3.76 br m60.4CH2.

[0107] The chemical structure of glucose-transferred ribaudioside A, in which 1 to 4 glucose molecules are added to the glucose connected to the 19th carbon of the above ribaudioside A via an α-(1,6) bond, is shown in FIG. 7. The specifications of the above glucose-transferred steviol glycoside are as follows. The pH range is between 4.5 and 7, the purity of the total steviol glycoside is 95% (w / w) or higher according to the JECFA 2021 test method, and the glucose-transferred steviol glycoside with alpha bonds was confirmed to be 95% (w / w) or higher based on the total steviol glycosides.

[0108] Example 1: Setting Equivalent Sweetness of Glucose-Transferred Steviol Glycosides

[0109] All sweeteners differ in the degree of sweetness expressed when actually dissolved in water. Therefore, the aqueous solution of the mixture of α-1,6 glucose-transferred steviol glycosides with 1 to 11 glucose transfers prepared in Example 1 (hereinafter A16) and the aqueous solution of a commercially available α-1,4 glucose-transferred steviol glycoside used as a conventional sweetener (Daepyung Co., hereinafter A14) were set to exhibit the same degree of sweetness expression.

[0110] First, to evaluate the sweetness of rebaudioside M (Reb M), the concentration of Reb M that exhibits an equivalent level of sweetness compared to a 10% aqueous sugar solution was analyzed.

[0111] Sweetness evaluation of Reb M

[0112] Specifically, the experiment was conducted using the 2-AFC (Two-alternative forced choice) method with 15 evaluators, and the number and proportion of responses that evaluated the sweetness as equivalent to the highest number for each concentration of Reb M (0.047%, 0.067%, and 0.107%) were measured to determine the concentration of Reb M that has the same sweetness as a 10% sugar solution. Meanwhile, d' (d-prime) in Table 5 below is a quantitative indicator representing the evaluator's sensitivity, and is a value representing the difference between the detected signal and the noise in units of standard deviation; the lower the value, the less difference there is between the two samples, and it can be calculated using the ratio of evaluator responses when the stimulus is presented versus when it is not presented.

[0113] Response Results (Number of Responses) Response Proportion Concentration (%) REB M Sugar 10% REB M Sugar 10% d'(fundamental measure of sensory difference)Conc 10.04 76 940.00% 60.00% 0.36Conc 20.06 75 1033.33% 66.67% 0.61Conc 30.10 710 566.67% 33.33% 0.61* Total number of trials = 15

[0114] When a beverage is prepared based on the sweetness of the results in Table 5 above, the concentration of Reb M corresponding to 8SEV (8% sugar aqueous solution) suitable for the beverage is calculated to be 0.0696%. Next, a comparative experiment on the advantages of sweetness using the 2-AFC (Two-alternative forced choice) method was conducted for the above A16 and Reb M at a concentration of 0.0696%. The 2-AFC experiment was performed on A16 at various concentrations (0.0931% and 0.1463%) to determine the concentration of A16 that has the same sweetness as Reb M at a concentration of 0.0696% corresponding to 8SEV.

[0115] Response Results (Number) Response Proportion Concentration (%) Reb MA16 REB MA16 d'Conc 10.09 316 634 66% 34% 0.82 Conc 20.14 6329 71 29% 71% 1.11* Total number of trials = 100

[0116] As a result, as shown in Table 6, the concentration of the glucose-transferred steviol glycoside (A16) corresponding to the above 8SEV was estimated to be 0.12%, and the sweetness was calculated to be 66.8 times that of sugar.

[0117]

[0118] Example 2: Setting Equivalent Sweetness for α-1,4 Glucose-Transferred Steviol Glycosides

[0119] In addition, the concentration of an α-1,4 glucose-transferred steviol glycoside (A14) having equivalent sweetness to 8SEV was analyzed. Based on the 0.0696% concentration of Reb M confirmed in Example 1, a comparative test of the sweetness advantage using the 2-AFC method was conducted. Specifically, 2-AFC experiments were performed on A14 at various concentrations (0.2973% and 0.3823%) to determine the concentration of A14 having the same sweetness as the 0.0696% concentration of Reb M corresponding to 8SEV.

[0120] Response Results (Number) Response Proportion Concentration (%) Reb MA14 REB MA14 d'Conc 10.29 734 357 43% 57% 0.35 Conc 20.38 232 47 624% 76% 1.41* Total number of trials = 100

[0121] Based on Table 7, the concentration of A14 corresponding to 8SEV was confirmed to be 0.25%, and the sweetness at this time was calculated to be 32.6 times that of sugar.

[0122]

[0123] Example 3: Sensory evaluation of a beverage product made using glucose-transferred steviol glycosides

[0124] 3-1. Sensory Evaluation of Cola Beverages

[0125] A sensory evaluation was performed to compare and analyze the sweetness of the beverage prepared using A16. For this purpose, a beverage prepared using A14 at the same concentration was used as a control group. Specifically, to prepare a cola beverage containing the above A16 and A14, a syrup was first prepared. The syrup was composed at a concentration five times that of the final cola beverage. The mixing ratio of the cola syrup was prepared as shown in Table 8 below. After accurately weighing each ingredient according to the mixing ratio, the final volume was adjusted with purified water, and the mixture was stirred sufficiently until all ingredients were completely dissolved.

[0126] In one embodiment, after preparing the cola syrup, it was diluted with carbonated water to a target concentration and utilized as a final Ready-to-Drink (RTD) beverage sample. To prepare the RTD beverage, 20% by weight of the syrup was mixed with 80% by weight of carbonated water and homogenized. In another embodiment, after preparing the syrup, it was mixed with 80% by weight of purified water, homogenized, and then carbonation was injected to produce a final RTD beverage sample. The carbonation pressure was 2.5 kg / cm². 2 This is desirable but not limited to this.

[0127] The input concentrations of A16 and A14 were applied at 66.8 times and 32.6 times, respectively, the sweetness values ​​calculated in Examples 1 and 2. For the diluted RTD cola beverage, the sweetness achieved using A16 and A14 was set to 8 SEV, and the sweetness achieved using allulose, an auxiliary sweetener, was set to 3 SEV, so that the sweetness of the final cola beverage was 11 SEV, thereby producing the final RTD cola beverage.

[0128] Ingredients A16 A14 A16 0.5985% -A14 -1.2270%Allulose 28.5500%28.5500%Cola Flavor 0.6000%0.6000%Others*Remaining Amount Total Remaining Amount 100.0000%100.0000%

[0129] Others: Caffeine, caramel coloring, citric acid, phosphoric acid, trisodium citrate, purified water, etc.

[0130] Subsequently, the above RTD beverage was provided to a sensory evaluation panel, or 20% by weight of purified water was mixed and homogenized, followed by carbonation. The sensory evaluation was conducted with 100 panelists familiar with taste quality evaluation. The beverage samples were evaluated using random 3-digit numbers to ensure there was no bias. As a result, as shown in Table 9 below, it was confirmed that the overall preference for the cola beverage containing A16 was higher than that of the cola beverage containing A14. Accordingly, it was confirmed that A16 exhibits excellent taste quality in actual beverage applications.

[0131] Classification Sweetness Intensity Flavor Intensity Overall Preference A16A14A16A14A16A14Avg 6.17 6.22 1.76 1.99 5.65 5.31

[0132] * bolded number means the result of a significant difference (p<0.05) for each evaluation.

[0133] 3-2. Sensory Evaluation of Lemon-Lime Carbonated Beverages

[0134] A lemon-lime flavored carbonated beverage base was syrupized in the same manner as in Example 3-1 above to prepare the beverage, and a sensory evaluation was performed. The mixing ratio of the lemon-lime beverage syrup is as shown in Table 10 below.

[0135] Ingredients A16 A14 A16 0.5985% A14 1.2270% Citric acid 0.3500% 0.3500% Others* Remaining amount Total remaining amount 100.0000% 100.0000%

[0136] *Others: Lemon / lime flavor, purified water, etc.

[0137] Sensory evaluation was conducted on a panel of 60 people familiar with taste quality evaluation.

[0138] As a result, as shown in Table 11 below, it was confirmed that the preference for lemon-lime carbonated beverages containing A16 was higher than that for lemon-lime carbonated beverages containing A14. Accordingly, it was confirmed that A16 exhibits excellent taste quality in actual beverage applications.

[0139] Classification Sweetness Intensity Flavor Intensity Overall Preference A16A14A16A14A16A14Avg 6.126.132.553.536.355.45

[0140] *bolded numbermeans the result of a significant difference (p<0.05) for each evaluation.

[0141] 3-3. Sensory Evaluation of Fruit and Vegetable Drinks (1)

[0142] An apple beverage was prepared by syruping in the same manner as in Example 3-1 above, and a sensory evaluation was performed. At this time, A16 and A14 were used at a concentration corresponding to 6 SEV in the final RTD beverage stage. The mixing ratio of the apple beverage syrup is as shown in Table 12 below. The apple juice concentrate was prepared to have a juice content of 50% (based on Single Brix 10) based on the total weight of the final RTD beverage, but the range of the juice content of the apple juice concentrate is not limited to this.

[0143] Ingredients A16 A14 Apple Concentrate 35.7000% 35.7000% A16 0.2090% A14 0.4285% Citric acid 0.5000% 0.5000% Others* Remaining Amount Total Remaining Amount 100.0000% 100.0000%

[0144] *Others: Malic acid, trisodium citrate, apple flavor, purified water, etc.

[0145] The above sensory evaluation was conducted on 30 panelists familiar with taste quality evaluation.

[0146] As a result, as shown in Table 13 below, it was confirmed that the preference for the apple beverage containing A16 was higher than that for the apple beverage containing A14. Accordingly, it was confirmed that A16 exhibits excellent taste quality in actual beverage applications.

[0147] Category Sweetness Intensity Bitterness Intensity Flavor Intensity Overall Preference A16A14A16A14A16A14A16A14A16A14Avg6.036.081.932.532.203.036.385.83

[0148] *bolded numbermeans the result of a significant difference (p<0.05) for each evaluation.

[0149] 3-4. Sensory Evaluation of Fruit and Vegetable Drinks (2)

[0150] Lemonade beverages were prepared by syruping the base in the same manner as in Example 3-3 above, and sensory evaluations were performed. In the final RTD beverage stage, A16 and A14 were used at a concentration corresponding to 8.4 SEV. To make lemonade, syrup is first prepared. The syrup is prepared at a concentration five times higher, considering the final beverage concentration at 1 / 5 of the level. The mixing ratio of the lemonade syrup is as shown in Table 14 below. Lemon juice extract (Lemon Juice (NFC)) was prepared to have a juice content of 17% (based on Single Brix 9) based on the total weight of the final RTD beverage, but the range of juice content of the juice extract is not limited to this.

[0151] Ingredients A16 A14 Lemon Juice (NFC) 85.0000% 85.0000% A16 0.6000% A14 1.2300% Others* Remaining Amount Total Remaining Amount 100.0000% 100.0000%

[0152] *Others: Lemon flavor, purified water, etc.

[0153] The above sensory evaluation was conducted on a panel of 100 people familiar with taste quality evaluation.

[0154] As a result, as shown in Table 15 below, it was confirmed that the overall preference of the lemonade beverage containing A16 was higher than that of the lemonade beverage containing A14. Accordingly, it was confirmed that A16 exhibits excellent taste quality in actual beverage applications.

[0155] Category Sweetness Intensity Overall Preference A16 A14 A16 A14 Avg 5.5 15.4 9 5.6 8 5.38

[0156] *bolded numbermeans the result of a significant difference (p<0.05) for each evaluation.

[0157]

[0158] Example 4: Mixture of glucose-transferred steviol glycosides and organic acids

[0159] The improved taste quality of the beverage when A16 and organic acids were mixed was investigated.

[0160] Specifically, acidic aqueous solutions with various pH conditions were prepared by adding organic acids widely used in carbonated beverages or various RTD beverages. The preparation of the acidic aqueous solutions was carried out by simplifying the mixing ratios. This was done to evaluate the taste quality of the glucose-transferred steviol glycoside of the present application in an environment where variables that could affect the taste quality of the acidic aqueous solution could be controlled in various organic acid application environments. Subsequently, to evaluate the taste quality of A16 in the acidic aqueous solution, 0.1% organic acid (citric acid, malic acid, tartaric acid, and phosphoric acid) and 0.1% beverage flavor (cola flavor, lemon flavor, complex citrus flavor), respectively, were mixed in purified water and mixed for 20 minutes using a magnetic mixer. Afterward, sweeteners were added to A16 at a concentration of 0.12% to adjust the sweetness to 8 SEV, and the mixture was further mixed and dissolved for 30 minutes to prepare tasting samples. After the preparation of the samples was completed, they were placed in a designated container and used for sensory evaluation. The above sensory evaluation evaluated the sourness intensity, flavor preference, and overall preference of the mixture sample and performed statistical analysis. At this time, as a comparison group, a mixture of A14, organic acid, and flavoring was used, with A14 added at a concentration of 0.25% to achieve the same sweetness of 8 SEV.

[0161] As a result, it was confirmed that the mixture of A16 and organic acid reduced sourness in a specific pH range and increased flavor preference and overall preference compared to the mixture of A14 and organic acid.

[0162] 4-1. Evaluation of A16 Validity of Cola-Flavored Acidic (Phosphoric Acid) Beverage Bases

[0163] Sample solutions having pH values ​​of 2.64, 3.18, and 3.83 were prepared by using A16 of the present application as described in the above manufacturing method, using phosphoric acid among the types of organic acids added, and adjusting the composition and content of the raw materials as shown in Tables 16 to 18.

[0164] IngrtdientsA16A14A160.1197% A14 0.2454%Allulose5.7100%5.7100%Caffeine0.0130%0.0130%Caramel color0.1400%0.1400%Phosphoric Acid0.0396%0.0390%Cola Flavor0.1200%0.1200%Purified water93.8577%93.7326%합계100.0000%100.0000%

[0165] IngrtdientsA16A14A160.1197% A14 0.2454%Allulose5.7100%5.7100%Caffeine0.0130%0.0130%Caramel color0.1400%0.1400%Phosphoric Acid0.0093%0.0093%Cola Flavor0.1200%0.1200%Purified water93.8880%93.7623%합계100.0000%100.0000%

[0166] IngrtdientsA16A14A160.1197% A14 0.2454%Allulose5.7100%5.7100%Caffeine0.0130%0.0130%Caramel color0.1400%0.1400%Phosphoric Acid0.0022%0.0022%Cola Flavor0.1200%0.1200%Purified water93.8951%93.7694%합계100.0000%100.0000%

[0167] In this case, a sample solution prepared using A14 was used as the control group. Sensory evaluations were conducted using samples with pH values ​​of 2.64, 3.18, and 3.83 to which the aforementioned phosphoric acid was applied. The efficacy verification evaluation was conducted with 50 sensory evaluation panelists who are typically trained in taste quality assessment. The evaluation was performed on a total scale of 9 points, with the evaluators assessing the samples treated with A16 and A14 in the domains of sourness intensity, flavor preference, and overall preference. Statistical analysis was performed using paired t-tests at a 95% confidence level to verify the significance of the two samples; a p-value less than 0.05 was considered to indicate a significant difference in evaluation results between the two samples. As shown in Table 19, the efficacy of A16 was confirmed, with sourness decreasing statistically and flavor preference increasing in the pH 3.18 range. Furthermore, a tendency was observed across all pH ranges where overall preference using A16 was significantly higher compared to A14.

[0168] Classification Sourness Intensity Cola Flavor Preference Overall Preference pHA 16A14A16A14A16A14 Low Concentration Average 2.18 2.2 23.9 0 3.6 25.4 6 5.18 3.83 p-value 0.35 0.05 40.07 Medium Concentration Average 2.85 3.3 13.9 6 3.5 6 4.8 0 4.3 7 3.18 p-value 0.03 0.01 0.03 High Concentration Average 3.89 4.3 44.9 24.6 35.4 8 5.10 2.64 p-value 0.02 50.09 80.07

[0169] 4-2. Evaluation of A16 Validity of Lemon-Flavored Acidic (Citric Acid) Beverage Bases

[0170] Next, citric acid was used among the types of organic acids added, and the composition and content of the raw materials were adjusted as shown in Tables 20 to 22 to prepare sample solutions having pH values ​​of 2.66, 3.17, and 3.81, respectively, with 0.1% lemon flavor added. Sensory evaluation was performed using the samples with pH values ​​of 2.66, 3.17, and 3.81 to which the above citric acid was applied. The sensory evaluation of taste quality was conducted in the same manner as in Example 4-1 described above.

[0171] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.1700%0.1700%salt0.0800%0.0800%Sodium magnesium0.0050%0.0050%Vitamin B20.000025%0.000025%Lemon Flavor0.1000%0.1000%Purified water99.5253%99.3996%Total100.0000%100.0000%

[0172] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.0240%0.0240%salt0.0800%0.0800%Sodium magnesium0.0050%0.0050%Vitamin B20.000025%0.000025%Lemon Flavor0.1000%0.1000%Purified water99.6713%99.5456%Total100.0000%100.0000%

[0173] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.0032%0.0032%salt0.0800%0.0800%Sodium magnesium0.0050%0.0050%Vitamin B20.000025%0.000025%Lemon Flavor0.1000%0.1000%Purified water99.6921%99.5664%Total100.0000%100.0000%

[0174] As shown in Table 23, the evaluation results confirmed the effect of A16, with a statistically significant decrease in sourness and an increase in flavor preference in the pH 3.17 range. A tendency was observed in all pH ranges where overall preference using A16 was significantly higher than that using A14.

[0175] Classification Sourness Intensity Lemon Flavor Preference Overall Preference pHA 16A14A16A14A16A14 Low Concentration Average 2.86 2.81 3.41 3.26 4.78 4.45 3.81 p-value 0.39 0.24 40.04 Medium Concentration Average 3.77 4.25 4.29 3.83 5.18 4.64 3.17 p-value 0.01 20.026 0.01 High Concentration Average 4.94 5.13 3.94 3.63 5.48 4.64 2.66 p-value 0.19 30.07 0.00 02

[0176] 4-3. Evaluation of A16 Validity of Acidic (Citric Acid) Beverage Bases with Complex Citrus Flavors

[0177] In addition, citric acid was used among the types of organic acids added, and the composition and content of the raw materials were adjusted as shown in Tables 24 to 26 to prepare sample solutions having pH values ​​of 2.51, 2.80, and 4.35, respectively, with 0.1% of complex citrus flavor added. Sensory evaluation was performed using the samples with pH values ​​of 2.51, 2.80, and 4.35 to which the above citric acid was applied. The sensory evaluation of taste quality was conducted in the same manner as in Example 4-1 described above.

[0178] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.3000%0.3000%salt0.0030%0.0030%Vitamin C0.0100%0.0100%Potassium Hydroxide0.0100%0.0100%Lemon-cola Flavor(Mt.dew Type)0.1000%0.1000%Purified water99.4573%99.3316%Total100.0000%100.0000%

[0179] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.0900%0.0900%salt0.0030%0.0030%Vitamin C0.0100%0.0100%Potassium Hydroxide0.0100%0.0100%Lemon-cola Flavor(Mt.dew Type)0.1000%0.1000%Purified water99.6673%99.5416%Total100.0000%100.0000%

[0180] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.0010%0.0010%salt0.0030%0.0030%Vitamin C0.0100%0.0100%Potassium Hydroxide0.0100%0.0100%Lemon-cola Flavor(Mt.dew Type)0.1000%0.1000%Purified water99.7563%99.6306%Total100.0000%100.0000%

[0181] As shown in Table 27, the evaluation results confirmed the effect of A16, with a statistically significant decrease in sourness and an increase in flavor preference in the pH 2.80 range. A tendency was observed in all pH ranges where overall preference using A16 was significantly higher than that using A14.

[0182] Classification Sourness Intensity Complex Citrus Aroma Overall Preference pHA 16A14A16A14A16A14 Low Concentration Average 1.73 1.763 3.383 1.44 53 4.364 3.5 p-value 0.41 0.063 0.15 Medium Concentration Average 3.62 4.22 4.63 4.125 54 5.162 8 p-value 0.001 0.004 0.021 High Concentration Average 5.165 3.53 3.45 5.585 13.25 1 p-value 0.12 0.32 0.009

[0183] 4-4. Evaluation of Unconfirmed Efficacy of A16 in Acidic Beverage Bases

[0184] Furthermore, by adjusting the content of tartaric acid and malic acid among the added organic acids, a sample solution was prepared that exhibited characteristics in the pH 3 range, which showed excellent effects in the flavor-applied organic acid liquid bases of Examples 4-1 to 4-3. The sample utilized a cola flavor and was prepared to have a pH of 3.10 using tartaric acid and a pH of 3.5 using malic acid, respectively. Sensory evaluation was performed using the samples to which each of these two types of organic acids was applied. The sensory evaluation of taste quality was conducted in the same manner as described in Example 4-1. The results for the sample using tartaric acid are summarized in Table 28 below, and the results for the sample using malic acid are summarized in Table 29 below.

[0185] As shown in Tables 28 and 29, the evaluation results indicated that although the acidic aqueous solutions to which the two organic acids were applied were both in the pH range of around 3, no statistical improvement in flavor preference was observed. This suggests that the effect of A16 does not appear in a specific pH range, but rather that the effect appears in the specific pH range of specific organic acids. In other words, when phosphoric acid and citric acid, which are widely used organic acids in acidic soft drinks, were used, the improvement in flavor preference of A16 was observed in the pH range of around 3; however, when organic acids such as tartaric acid and malic acid were used, no effect was observed. This indicates that when specific types of organic acids are used, the acidity of the beverage is reduced and the flavor is improved in a specific pH range. Meanwhile, there was a tendency for the overall preference to be significantly higher when using A16 compared to A14.

[0186] pH 3.1 Sourness Intensity Aroma Overall Preference A16 A14 A16 A14 A16 A14 Evaluation Score 2.90 2.92 5.30 5.52 5.98 5.46 p-Value p>0.05 p>0.05 p<0.05

[0187] pH 3.5 Sourness Intensity Aroma Preference Overall Preference A16 A14 A16 A14 A16 A14 Evaluation Score 3.6 4 3.9 0 4.2 8 4.1 4 6.3 2 5.26 p-Value p>0.05 p>0.05 p<0.05

[0188] 4-5. Evaluation of the Valid pH Range of A16 in Cola-Flavored Acidic Beverage Bases

[0189] In Example 4-1 above, the efficacy of reducing acidity and enhancing flavor preference of the cola-flavored beverage base using phosphoric acid and A16 was confirmed, so additional experiments were conducted to determine a more specific pH range. As the pH range of cola investigated in Example 4-1 was found to be between 2.7 and 3.5, to verify the effect within this pH range, sample solutions exhibiting pH values ​​of 2.7 and 3.5 were prepared and produced by adjusting the composition and content of the raw materials using phosphoric acid as shown in Tables 30 to 31, respectively, and then a sensory evaluation of the taste quality of the samples was conducted. The sensory evaluation method was the same as described in Example 4-1 above, and the evaluation panel consisted of 30 people.

[0190] IngrtdientsA16A14A160.1197% A14 0.2454%Allulose5.7100%5.7100%Caffeine0.0130%0.0130%Caramel color0.1400%0.1400%Phosphoric Acid0.0250%0.0250%Cola Flavor0.1200%0.1200%Purified water93.8723%93.7466%Total100.0000%100.0000%

[0191] IngrtdientsA16A14A160.1197% A14 0.2454%Allulose5.7100%5.7100%Caffeine0.0130%0.0130%Caramel color0.1400%0.1400%Phosphoric Acid0.0050%0.0050%Cola Flavor0.1200%0.1200%Purified water93.8923%93.7666%Total100.0000%100.0000%

[0192] As a result, as shown in Table 32, flavor improvement effects were confirmed in the pH 2.7 and 3.5 ranges. Through this, it was found that acidic beverage products treated with phosphoric acid and A16 have excellent sensory quality when the pH range is 2.7 to 3.5.

[0193] Category (pH 2.7) Sourness Intensity Flavor Preference Overall Preference A16 A14 A16 A14 A16 A14 Evaluation Score 3.10 3.5 73.8 73.30 4.8 34.41 p-Value p<0.05 p<0.05 p<0.05 Category (pH 3.5) Sourness Intensity Flavor Preference Overall Preference A16 A14 A16 A14 A16 A14 Evaluation Score 2.80 3.30 4.2 73.7 35.4 74.82 p-Value p<0.05 p<0.05 p<0.05

[0194]

[0195] 4-6. Evaluation of the Valid pH Range of A16 for Lemon-Flavored Acidic Beverage Base

[0196] In Example 4-2 above, the effectiveness of reducing acidity and enhancing flavor preference of the lemon-flavored beverage base using citric acid and A16 was confirmed, so additional experiments were conducted to determine a more specific pH range. As a result of investigating the pH range of the lemon-flavored beverage in Example 4-2, it was confirmed to be at the pH level of 2.7 to 3.5. To verify the effect in this range, citric acid was used to adjust the composition and content of the raw materials as shown in Tables 33 to 34, and sample solutions exhibiting pH values ​​of 2.9 and 3.3, respectively, were prepared and produced, after which a sensory evaluation of the taste quality of the samples was conducted. The sensory evaluation method is the same as that described in Example 4-5 above.

[0197] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.0620%0.0620%salt0.0800%0.0800%Sodium magnesium0.0050%0.0050%Vitamin B20.000025%0.000025%Lemon Flavor0.1000%0.1000%Purified water99.6333%99.5076%Total100.0000%100.0000%

[0198] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.0150%0.0150%salt0.0800%0.0800%Sodium magnesium0.0050%0.0050%Vitamin B20.000025%0.000025%Lemon Flavor0.1000%0.1000%Purified water99.6803%99.5546%Total100.0000%100.0000%

[0199] As a result, as shown in Table 35, flavor improvement effects were confirmed in the pH 2.9 and pH 3.3 ranges. Through this, it was found that acidic beverage products to which citric acid and A16 were applied have excellent sensory quality when the pH range is 2.9 to 3.3.

[0200] Classification Sourness Intensity Flavor Overall Preference A16A14A16A14A16A14 Evaluation Score 3.07 3.77 4.43 3.90 4.97 4.13 p-Value p<0.05 p<0.05 p<0.05 Classification Sourness Intensity Flavor Overall Preference A16A14A16A14A16A14 Evaluation Score 2.60 3.13 4.23 3.73 6.23 5.40

[0201]

[0202] 4-7. Evaluation of A16 Effective pH Range for Acidic Beverage Bases with Complex Citrus Flavors

[0203] In Example 4-3, another example in which the above citric acid was applied, the effectiveness of reducing acidity and enhancing flavor preference of the citrus-flavored acidic beverage base to which citric acid and A16 were applied was confirmed; therefore, additional experiments were conducted to determine a more specific pH range. In Example 4-3, the pH range of the complex citrus-flavored beverage was investigated and found to be between 2.8 and 4.0, while the effect of improving flavor preference at pH 2.8 was confirmed. Therefore, a sample solution exhibiting a pH of 4.0 was prepared and produced by adjusting the composition and content of the raw materials using citric acid as shown in Table 36, and then the taste quality of the sample was evaluated. The sensory evaluation method is the same as that described in Example 4-5.

[0204] IngrtdientsA16A14A160.1197% A14 0.2454%Citric acid0.0019%0.0019%salt0.0030%0.0030%Vitamin C0.0100%0.0100%Potassium Hydroxide0.0100%0.0100%Lemon-cola Flavor(Mt.dew Type)0.1000%0.1000%Purified water99.7554%99.6297%Total100.0000%100.0000%

[0205] As a result, as shown in Table 37, it was confirmed that the flavor improvement effect was observed even in the pH 4.0 range. Through this, it was found that the complex citrus flavored acidic beverage product containing citric acid and A16 has excellent sensory quality when the pH range is 2.8 to 4.0.

[0206] Category (pH 4.0) Sourness Intensity Flavor Preference Overall Preference A16 A14 A16 A14 A16 A14 Evaluation Score 3.70 4.20 5.00 4.20 5.37 4.73 p-Value p<0.05 p<0.05 p<0.05

[0207]

[0208] Although representative embodiments of the present application have been described above by way of example, the scope of the present application is not limited to such specific embodiments, and those skilled in the art will be able to make appropriate modifications within the scope described in the claims of the present application.

[0209]

[0210] [Consignment Number]

[0211] Name of depositing institution: Korean Culture Collection of Microorganisms

[0212] Trustee Number: KCCM13503P

[0213] Date of Trust: 20240805

[0214]

Claims

A composition comprising a glucose-transferred steviol glycoside and an organic acid, The above-mentioned steviol glycoside comprises at least one selected from the group consisting of stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside O, rebaudioside N, rebaudioside I, rebaudioside M, lubusoside, steviolbioside, and dulcoside A. A composition in which the glucose-transferred steviol glycoside comprises one or more glucose molecules added via an α-1,6 bond to a glucose molecule connected to the 19th carbon of the steviol glycoside. In claim 1, A composition in which the glucose-transferred steviol glycoside comprises 1 to 11 glucose molecules added via α-1,6 bonds to a glucose molecule connected to the 19th carbon of the steviol glycoside. In claim 1, A composition wherein the organic acid is at least one selected from the group consisting of phosphoric acid, citric acid, malic acid, tartaric acid, lactic acid, formic acid, ascorbic acid, fumaric acid, gluconic acid, succinic acid, maleic acid, and adipic acid. In claim 1, A composition in which the above organic acid is at least one selected from the group consisting of phosphoric acid, citric acid, malic acid, and tartaric acid. In claim 1, A composition in which the content ratio of the glucose-transferred steviol glycoside and the organic acid is 0.001 : 0.5 to 1 : 0.0001. In claim 1, The above composition is a beverage composition. In claim 6, A composition comprising the glucose-transferred steviol glycoside in an amount of 0.001 to 1 weight part based on 100 weight parts of the beverage. In claim 6, A composition in which the above organic acid is included in an amount of 0.0001 to 0.5 parts by weight based on 100 parts by weight of the beverage. A beverage comprising the composition of any one of claims 1 to 8. In claim 9, A beverage having a content of 0.001 to 1 part by weight of the glucose-transferred steviol glycoside based on 100 parts by weight of the beverage. In claim 9, A beverage having a content of 0.0001 to 0.5 parts by weight of the organic acid based on 100 parts by weight of the beverage. In claim 9, A beverage having a pH of 2.0 to 5.0.

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