Composition comprising glucosylated steviol glycosides

Abstract

The present application relates to a food composition comprising glucosylated steviol glycosides, and to foods comprising same. Foods prepared by using the food composition comprising the glucosylated steviol glycosides of the present application exhibit excellent taste quality in terms of the intensity of sweetness and off-flavor, resulting in improved overall palatability.

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. 10-2025-0012716 filed on January 31, 2025, Korean Patent Application No. 10-2025-0119488 filed on August 26, 2025, and Korean Patent Application No. 10-2025-0212676 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 food composition comprising a glucose-transferred steviol glycoside and a food 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 astringency, 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 food odors and off-flavors by utilizing glucose-transferred stevia glycosides, and to provide a food composition and food containing said glucose-transferred stevia glycosides.

[0017]

[0018] One aspect of the present application provides a food composition comprising a glucose-transferred steviol glycoside and a bulk sweetener, 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 a glucose atom connected to the 19th carbon of the steviol glycoside via an α-1,6 bond.

[0019] Another aspect of the present application provides a food comprising the above food 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, 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 mali KCCM13503P (Lactobacillus mali CJST242), DSM20444, ATCC 27054, or ATCC 27304.

[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 food composition comprising the glucose-transferred stevia glycoside and a bulk sweetener.

[0035] The food compositions of the present application include, but are not limited to, general food, health food, and medical (or patient) food compositions.

[0036] In addition, the above food composition may be an additive or supplement contained in food and beverages.

[0037] The above food may include all ingredients or combinations of ingredients or all substances or combinations of substances that can be used as food for mammals or prepared for use as food for mammals, and may include substances that can be used in the manufacture of food or food additives, but are not limited thereto. The food composition of the present application may be applied to various foods requiring sweetness, for example, foods to which the composition may be applied include beverages (e.g., dietary fiber drinks, soft drinks, carbonated water, misugaru, coffee drinks, yogurt drinks, lattes, coffee milk, soy milk, milk-containing drinks, etc.), juices (e.g., fruit juices, fruit and vegetable juices, etc.), bakery products (e.g., cookies, cakes, hotcakes, pies, brownies, bread, sliced ​​bread, crackers, etc.), sweetened toppings (e.g., icing, whipped cream, etc.), jellies, chewing gum, jams, snack foods (e.g., cookies, potato chips, tortilla chips, popcorn, snack bars, etc.), grain-based foods (e.g., oats, oatmeal, cereals, rice cakes, granola bars, etc.), sauces (e.g., tonkatsu sauce, ketchup, mayonnaise, ranch sauce, teriyaki sauce, mala tang sauce, salad dressing, etc.), dairy products (e.g., fermented milk, etc.). Fermented foods (e.g., napa cabbage kimchi, gochujang, etc.), pickled foods (e.g., pickled radish, pickled vegetables, etc.), braised dishes (e.g., braised quail eggs, braised mackerel, braised baby potatoes, braised black beans, braised saury in soy sauce, etc.), beef rice bowls, stir-fries (e.g., stir-fried fish cakes, stir-fried eggplant, stir-fried anchovies, stir-fried squid, stir-fried dried squid strips, stir-fried vegetables, and stir-fried beef, etc.), seasoned salads (e.g., seasoned raw vegetables, seasoned mature cucumbers, seasoned pickled cucumbers, and seasoned seasoned greens, etc.), grilled dishes (e.g., grilled squid, grilled LA galbi, grilled rice cakes, etc.), soups and broths (e.g., stews, soups, broths, etc.), syrups, dressings, tteokbokki, beef bulgogi, steamed Korean chili peppers, kiwi syrup, ssamjang, acidulants, pharmaceutical preparations, infant formula, infused foods (e.g., fruits and vegetables, etc.), seasonings, syrups, desserts (e.g., Candy, hard candy, chocolate, dark chocolate, hotteok, pudding,Frozen desserts such as ice cream and sherbet, etc.), soft frozen products (e.g., soft frozen cream, soft frozen yogurt, soft frozen toppings such as dairy or non-dairy whipped toppings, etc.), toothpaste, mouthwash, oil, emulsified products (e.g., shortening, margarine, mayonnaise, butter, cooking oil, etc.), medium-moisture foods (e.g., rice, animal feed, etc.), or processed foods may be included, but are not limited thereto, and the above foods may be foods according to the classification criteria of the Food Code under the Food Sanitation Act.

[0038] 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, yogurt 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 beverage 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.

[0039] Examples of the above juice sources may include, but are not limited to, 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, 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, and lingonberries. No. Numerous additional and alternative juices suitable for use in at least certain embodiments will be apparent to those skilled in the art, taking into account the benefits of this application. Furthermore, the specific amount of juice useful for imparting flavor characteristics to the beverage product may vary depending on the selected juice, the desired flavor impression, and the form of the juice ingredients. Taking into account the benefits of this application, 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.

[0040] In one embodiment, the food composition of the present application may be used to produce at least one food selected from the group consisting of coffee beverages, yogurt beverages, soy milk beverages, cookies, chocolates, candies, and chewing gums.

[0041] In addition, the food composition of the present application can be utilized in various specifications for intermediate raw materials such as semi-finished food products or frozen dough.

[0042] In various foods, the content of the glucose-transferred steviol glycoside of the food composition of the present application may be 0.0001 to 3 parts by weight based on 100 parts by weight of the food, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 0.0001, 0.0005, 0.001, 0.005, 0.007, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, and 0.07 parts by weight, and an upper limit selected from the group consisting of 0.12, 0.15, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, and 3 parts by weight, for example, the content of the glucose-transferred steviol glycoside is 100 Based on weight parts, it may be 0.0005 to 2 weight parts, 0.001 to 1 weight part, 0.005 to 3 weight parts, 0.005 to 0.9 weight parts, 0.007 to 0.8 weight parts, 0.005 to 0.7 weight parts, 0.01 to 0.6 weight parts, 0.02 to 0.5 weight parts, 0.03 to 0.4 weight parts, 0.03 to 0.3 weight parts, 0.03 to 0.12 weight parts, 0.04 to 0.2 weight parts, 0.04 to 0.15 weight parts, 0.05 to 0.12 weight parts, or 0.07 to 0.12 weight parts.

[0043]

[0044] The above bulk sweetener may include sugar alcohols or sugars. The above sugar alcohols are substances formed by reducing the carbonyl group of sugars and may also be called sugar alcohols; for example, they may be erythritol, xylitol, arabitol, mannitol, sorbitol, maltitol, or lactitol, but are not limited thereto. Additionally, the above sugars may be at least one of monosaccharides, disaccharides, and oligosaccharides. The above monosaccharides may be, for example, arabinose, xylose, tagatose, allulose, allose, or galactose, and the above disaccharides are sugars formed by the combination of two monosaccharides; for example, they may be lactose, maltose, trehalose, turanose, or cellobiose, but are not limited thereto. , for example, in the present application, the bulk sweetener is erythritol, sorbitol, mannitol, xylitol, lactitol, isomalt, malitol, tagatose, trehalose, galactose, rhamnose, cyclodextrin, ribulose, threose, arabinose, xylose, lixos, allose, altroose, mannose, iidos, lactose, maltose, isotrehalose, neotrehalose, palatinose or isomaltulose, erythrose, deoxyribose, goulose, talose, erythrulose, xylluose, psicose, turanos, cellobiose, glucosamine, mannosamin, fucose, fuculose, glucuronic acid, gluconic acid, gluconolactone, abequeose, galactosamine, xylooligosaccharide (xyllotrios, It may include at least one selected from the group consisting of xylobiose, etc.), gentiooligosaccharides (gentiobiose, gentiotriose, gentiotetraose, etc.), galactooligosaccharides, sorbose, ketotriose (dehydroxyacetone), aldotriose (glyceraldehyde), nigerooligosaccharides, fructooligosaccharides (ketoose, nistose, etc.), maltotetraose, maltotriol, tetrasaccharides, mannanoligosaccharides, maltooligosaccharides (maltotriose, maltotetraose, maltopentase, maltohexaose, maltoheptase, etc.), dextrin, lactulose, melivise, raffinose, rhamnose, and ribose.

[0045] In one embodiment, the food composition of the present application may include the glucose-transferred steviol glycoside of the present application and xylitol and / or isomalt as a bulk sweetener.

[0046] In various foods, the content of the bulk sweetener in the food composition of the present application may be 1 to 99.99 parts by weight based on 100 parts by weight of the food, and furthermore, the content of the bulk sweetener useful for imparting sweetness characteristics to the food may vary depending on the type, form, taste, etc. of the selected food. Given the advantages of the present application, a person skilled in the art will be able to appropriately determine the content of the bulk sweetener used to achieve the desired flavor, taste quality, form, etc. Specifically, the content of the bulk sweetener in the food composition of the present application may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 1, 4, 10, 12, 14, 15, 20, 30, 38, 40, 49, 50, 60, 63, 70, 80, and 90 parts by weight based on 100 parts by weight of food, and an upper limit selected from the group consisting of 10, 12, 15, 16, 20, 30, 39, 40, 50, 60, 64, 70, 80, 90, 99.5, and 99.99 parts by weight; for example, the content of the bulk sweetener may be 1 to 99.5 parts by weight, 1 to 90 parts by weight, or 1 to 12 parts by weight based on 100 parts by weight of food. It may be parts by weight, 1 to 16 parts by weight, 4 to 15 parts by weight, 4 to 39 parts by weight, 10 to 20 parts by weight, 10 to 90 parts by weight, 12 to 64 parts by weight, 20 to 80 parts by weight, 20 to 70 parts by weight, 20 to 50 parts by weight, 30 to 60 parts by weight, 50 to 80 parts by weight, 60 to 70 parts by weight, 4 to 64 parts by weight, 14 to 50 parts by weight, or 90 to 99.99 parts by weight, but is not limited thereto.

[0047] When the above bulk sweetener is xylitol, the xylitol content may be 1 to 99.99 parts by weight based on 100 parts by weight of food, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 1, 4, 10, 12, 20, 30, 40, 49, 50, 60, 63, 70, 80, and 90 parts by weight, and an upper limit selected from the group consisting of 10, 12, 20, 30, 40, 50, 60, 64, 70, 80, 90, 99, 99.5, and 99.99 parts by weight; for example, the xylitol content may be 1 to 99.5 parts by weight, 1 to 90 parts by weight, or 10 to 20 parts by weight based on 100 parts by weight of food. It may be in parts by weight, 10 to 90 parts by weight, 20 to 80 parts by weight, 20 to 70 parts by weight, 20 to 50 parts by weight, 30 to 60 parts by weight, 50 to 80 parts by weight, 60 to 70 parts by weight, 4 to 64 parts by weight, or 90 to 99.99 parts by weight, but is not limited thereto.

[0048] When the above bulk sweetener is isomalt, the isomalt content may be 1 to 99.99 parts by weight based on 100 parts by weight of food, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 1, 10, 14, 15, 20, 30, 38, 40, 49, 50, 60, 70, 80, and 90 parts by weight, and an upper limit selected from the group consisting of 10, 15, 16, 20, 30, 39, 40, 50, 60, 70, 80, 90, 99, 99.5, and 99.99 parts by weight; for example, the isomalt content may be 1 to 99.5 parts by weight, 1 to 90 parts by weight, 10 to It may be 20 parts by weight, 10 to 90 parts by weight, 20 to 80 parts by weight, 20 to 70 parts by weight, 20 to 50 parts by weight, 30 to 60 parts by weight, 50 to 80 parts by weight, 60 to 70 parts by weight, 14 to 50 parts by weight, or 90 to 99.99 parts by weight, but is not limited thereto.

[0049] In addition to glucose-transferred steviol glycosides, the above food 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, D-Psicose, 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 foods, the content of the sweetener may be 0.01 to 90 parts by weight based on 100 parts by weight of the food, but is not limited thereto.

[0050] The food composition of the present application can be added to food as is, used together with other food ingredients, or used appropriately according to conventional methods.

[0051] The food composition of the present application may contain various additional ingredients that may be included in the food, depending on the type of food to be manufactured.

[0052] Specifically, the food composition of the present application may further include at least one additive selected from the group consisting of 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, flavoring agents, flavorings, coloring agents, carbohydrate carbonating agents, and acidifiers. The nutrients may include vitamins or minerals. The acidifier may include 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. The content of these additives may be selected in the range of 0.01 to 90 parts by weight based on 100 parts by weight of the food of the present application.

[0053]

[0054] Another aspect of the present application provides a food comprising the above food composition.

[0055] Based on 100 parts by weight of the above food, the content of the glucose-transferred steviol glycoside of the composition may be 0.0001 to 3 parts by weight, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 0.0001, 0.0005, 0.001, 0.005, 0.007, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, and 0.07 parts by weight, and an upper limit selected from the group consisting of 0.12, 0.15, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, and 3 parts by weight; for example, the content of the glucose-transferred steviol glycoside is based on 100 parts by weight of the food, It may be 0.0005 to 2 parts by weight, 0.001 to 1 part by weight, 0.005 to 3 parts by weight, 0.005 to 0.9 parts by weight, 0.007 to 0.8 parts by weight, 0.005 to 0.7 parts by weight, 0.01 to 0.6 parts by weight, 0.02 to 0.5 parts by weight, 0.03 to 0.4 parts by weight, 0.03 to 0.3 parts by weight, 0.03 to 0.12 parts by weight, 0.04 to 0.2 parts by weight, 0.04 to 0.15 parts by weight, 0.05 to 0.12 parts by weight, or 0.07 to 0.12 parts by weight.

[0056] The above food may further include at least one additive selected from the group consisting of nutritional supplements, 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, flavoring agents, fragrances, coloring agents, carbonating agents, and acidifiers. Based on 100 parts by weight of the above food, the content of the additive may be 0.01 to 90 parts by weight.

[0057] The description regarding the specific types or examples of the above-mentioned food can be understood as identical to what has been stated above, so it is not described redundantly.

[0058] The above food may be at least one selected from the group consisting of beverages, confectionery, baked goods, desserts, and candies.

[0059] The above food may be at least one selected from the group consisting of coffee beverages, yogurt beverages, cookies, chocolates, candies, and chewing gum.

[0060] In the case where the above food is a coffee beverage, the above food may further include, in addition to the glucose-transferred steviol glycoside, coffee bean extract, milk (skimmed milk powder, etc.), maltodextrin, coffee flavoring, milk flavoring, water, or purified water.

[0061] In the case where the above food is a yogurt beverage, the above food may further include raw yogurt, fruit puree (strawberry puree, etc.), milk, or flavoring (strawberry flavoring, etc.) in addition to the glucose-transferred steviol glycoside.

[0062] In the case where the above food is a cookie, the above food may further include butter, bulk sweetener, fresh cream, milk powder (skimmed milk powder, etc.), wheat flour, or cake flour in addition to the glucose-transferred steviol glycoside.

[0063] In the case where the above food is chocolate, the above food may further include a bulk sweetener, cocoa mass, cocoa butter, an emulsifier (lecithin, etc.), a flavoring (vanilla flavoring, milk flavoring, etc.) or vanillin in addition to the glucose-transferred steviol glycoside.

[0064] In the case where the above food is candy, the above food may further include bulk sweeteners, fruit extracts (lemon extract, etc.), acidulants (citric acid, etc.), flavorings (lemon flavoring, mint flavoring, etc.) or colorings in addition to the glucose-transferred steviol glycosides.

[0065] In the case where the above food is chewing gum, the above food may further include bulk sweeteners, gums, emulsifiers, or flavorings (such as apple mint flavoring) in addition to the glucose-transferred steviol glycoside.

[0066]

[0067] The glucose-transferred steviol glycoside of the present application has improved bitterness and sweetness palatability, and has excellent sweetness quality with low off-flavor or off-odor. When food is manufactured using a food composition containing such a glucose-transferred steviol glycoside, the off-flavor or off-odor of the steviol glycoside sweetener is improved, and excellent taste quality is exhibited in terms of sweetness intensity and off-flavor intensity, thereby increasing the overall palatability of the food. Thus, the food composition of the present application can be utilized in various food fields.

[0068] However, the effects of the present application are not limited to those mentioned above, and other unmentioned effects will be clearly understood by those skilled in the art from the following description.

[0069]

[0070] Figure 1 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside A by the Lactobacillus mali CJST242 strain.

[0071] Figure 2 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside C by the Lactobacillus mali CJST242 strain.

[0072] Figure 3 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside F by the Lactobacillus mali CJST242 strain.

[0073] Figure 4 is an HPLC chromatogram of the result of the glycosylation reaction of stevioside by the Lactobacillus mali CJST242 strain.

[0074] Figure 5 is an HPLC chromatogram of the result of the glycosylation reaction of dulcosides by the Lactobacillus mali CJST242 strain.

[0075] Figure 6 is an HPLC chromatogram of the glycosylation reaction results of rubusoside by the Lactobacillus mali CJST242 strain.

[0076] 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.

[0077]

[0078] The present application will be explained in more detail below through examples.

[0079] 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.

[0080]

[0081] Preparation Example 1: Preparation of glucose-transferred steviol glycosides

[0082] The preparation of glucose-transferred steviol glycosides was carried out according to the method described in Korean Patent Publication No. 10-2021-0114899.

[0083] 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.

[0084] 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 ribaudioside A, ribaudioside C, ribaudioside F, stevioside, dulcoside A, or rubusoside, and HPLC was used to confirm whether glucose-transferred stevioside, glucose-transferred rubusoside, glucose-transferred dulcoside A, or glucose-transferred ribaudioside A / C / F were produced (see 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.

[0085] 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.

[0086] 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).

[0087] The results are listed in Tables 1 to 4 below.

[0088] 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").

[0089] 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").

[0090] 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.

[0091] 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.

[0092] 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.

[0093] w Weak signals.

[0094] * The chemical shift of each signal may not be exact value due to signal overlapping

[0095] a,b,c,d,e,f,g,hThe assignment of signals at same alphabet column may be exchangeable.

[0096]

[0097] 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.

[0098] 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.

[0099] 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.

[0100] 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.

[0101]

[0102] Example 1: Setting Equivalent Sweetness of Glucose-Transferred Steviol Glycosides

[0103] 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.

[0104] 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.

[0105] Sweetness evaluation of Reb M

[0106] 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.

[0107] 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

[0108] 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.

[0109] 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

[0110] 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.

[0111]

[0112] Example 2: Setting Equivalent Sweetness for α-1,4 Glucose-Transferred Steviol Glycosides

[0113] 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.

[0114] 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

[0115] 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.

[0116]

[0117] Example 3: Sensory evaluation of a product made using glucose-transferred steviol glycosides

[0118] Sensory evaluation was performed to compare and analyze the sweetness of food prepared using A16 and food prepared using A14. Since the above A16 can reduce off-flavors or odors of food additives, the sweetness or taste quality of the food prepared using A16 was improved. The manufacturing process is as follows.

[0119] 3-1. Soy Milk Products

[0120] First, the raw materials were weighed according to the mixing ratios listed in the table below, and soy milk containing the above A16 and xylitol was prepared. The mixing ratio of the soy milk is as shown in Table 8 below. Specifically, soybeans were selected and washed, and inedible parts were removed. Subsequently, the beans were immersed in purified water to hydrate them, and a 10-fold increase in water was added. Wet grinding was then performed to obtain a soybean slurry. The obtained soybean slurry was heated to approximately 95 degrees Celsius to inactivate internal enzymes. This process is typically used to remove fishy odors and off-flavors; however, since consumers perceive the performance as poor, there is a need for fishy odor removal in many soy milk products. Subsequently, insoluble solids were removed through filtration and centrifugation processes. The separated filtrate was further heated and sterilized at over 90 degrees Celsius to eliminate the possibility of microbial contamination and proliferation. After adjusting the solid content concentration as necessary, the mixture was homogenized at a pressure of 10 to 30 MPa to obtain the soy milk liquid. A16 or A14 sweetener was added to the above soy milk liquid according to the formulation shown in Table 8 below. Each ingredient was weighed and prepared according to the specified ratio, and the prepared ingredients were added to the measured soy milk liquid and mixed and stirred. Once the mixing was complete, the mixture was sterilized at 95°C for 15 to 20 minutes, and then filled into sterilized packaging, such as standing pouches, in a sterile environment. Afterward, it was immediately cooled to 25°C and used as a sample for sensory evaluation.

[0121] Sensory evaluation was conducted on 30 well-trained panelists, and samples were provided with random 3-digit numbers to ensure there was no bias during evaluation. The sensory evaluation was performed on four items as shown in Table 9 below, and each sample was inverted 15 times to prevent bias based on the order of presentation. The results of the sensory evaluation for soy milk prepared using the above xylitol and A16 and A14 are shown in Table 9.

[0122] Ingredient Name | Mixing Ratio (%) A16 | A14 | Soy Milk Extract 93% | 93% Xylitol 4% | 4% A16 | 0.03% A14 | 0.06% Purified Water 2.97% | 2.94% Total 100% | 100%

[0123] Category Sweetness Intensity Bitterness Intensity Off-flavor Intensity (Fishy Odor) Overall Preference A16A14A16A14A16A14A16A14 Average 4.97 4.70 2.27 2.87 3.03 3.63 6.07 5.10 p-value p>0.05 p<0.05 p<0.05 p<0.05

[0124] As shown in Table 9, the evaluation results indicate that when A16 is added, the fishy odor of soy milk is suppressed and the bitterness is reduced, increasing overall preference, thus proving the superior taste quality of A16.

[0125] 3-2. Cookie Products

[0126] First, the raw materials were weighed according to the mixing ratios listed in the table below, and cookies containing the above A16 and xylitol or isomalt were prepared. The mixing ratios of the cookies are as shown in Tables 10 and 11 below. Specifically, butter was weighed and left at room temperature, then mixed with a high-speed mixer for 5 minutes to make a soft cream. Afterwards, fresh cream and xylitol or isomalt were weighed and added, and mixed for 10 minutes until they were sufficiently melted. Then, the weighed A16 was added, and mixed for another 5 minutes. Skim milk powder and cake flour were added and mixed according to the formula listed in the above cream-shaped molding product. The above mixture was placed in a bakery mixer and mixed for 20 minutes to make a dough, and the dough was placed in a refrigerator (at approximately 5 degrees Celsius) for 1 hour to mature, then divided into 3 to 15g portions, shaped into round shapes, and baked in an oven. The oven was preheated to 180 degrees Celsius, and the baking tray containing the cookie dough was placed inside and baked for 8 minutes. After baking, the cookies were removed from the oven and cooled at room temperature for 1 hour. At this time, A14 was used as the control group instead of A16, and cookies were prepared in the same manner. After preparing the cookies, they were used for sensory evaluation, which was conducted on a panel of 30 individuals familiar with taste quality evaluation. Samples were provided using random 3-digit numbers and in random order to ensure there was no bias during evaluation. The results of the sensory evaluation for the cookies prepared using xylitol are shown in Table 12, and the results of the sensory evaluation for the cookies prepared using isomalt are shown in Table 13.

[0127] Ingredient Mixing Ratio A16 A14 Butter 23.04% 23.04% Xylitol 12.11% 12.11% A16 0.05% A14 0.10% Fresh Cream 9.60% 9.55% Skim Milk Powder 7.20% 7.20% Cake Flour 48.00% 48.00% Total 100.00% 100.00%

[0128] Ingredient Mixing Ratio A16 A14 Butter 23.04% 23.04% Isomalt 15.50% 15.50% A16 0.07% A14 0.14% Fresh Cream 9.60% 9.60% Skim Milk Powder 3.79% 3.72% Cake Flour 48.00% 48.00% Total 100.00% 100.00%

[0129] As a result of conducting a sensory evaluation on the manufactured cookies, as shown in Tables 12 and 13, the cookies made with A16 showed better results in overall preference, with increased preference for off-flavor and cookie aroma in taste quality compared to A14.

[0130] Category Sweetness Intensity Ontaste Intensity Cookie Aroma Overall Preference A16A14A16A14A16A14A16A14 Evaluation Score 5.80 5.70 1.5 32.07 6.23 5.77 6.03 5.33 p-Value p>0.05p<0.05p<0.05p<0.05

[0131] Category Sweetness Intensity Ontaste Intensity Cookie Aroma Overall Preference A16A14)A16A14A16A14A16A14 Evaluation Score 5.6 35.3 31.6 0 2.1 0 5.9 35.1 0 6.4 35.6 7 p-Value p>0.05 p<0.05 p<0.05 p<0.05

[0132] 3-3. Candy Products

[0133] A sugar-free lemon mint candy was prepared with two layers, separated into a xylitol layer and an isomalt layer. To produce a candy with a hard texture, sugar alcohols such as isomalt and xylitol were used as bulk sweeteners. Meanwhile, to complement the sweetness of the sugar alcohols isomalt and xylitol, the above-mentioned A16 or A14 was used at a concentration corresponding to 6 SEV, utilizing the sweetness evaluation results described above. The specific mixing ratio of the hard candy is shown in Table 14 below. First, to prepare the above-mentioned isomalt layer, isomalt was dissolved in hot water at 100°C, and the solution was heated to a temperature of 167°C to prepare a candy mass. The mass was cooled to 140°C, and citric acid, flavoring, coloring, and A16 or A14 were added and poured into a mold. Next, to prepare the xylitol layer, xylitol was heated to 155°C to melt it into a liquid state, then cooled to 90°C and poured onto a mold filled with the isomalt mixture. After cooling to 30°C, it was removed from the mold to produce a two-layer hard candy. Sensory evaluation was conducted on the hard candy samples prepared as described above, and the evaluation was performed on a panel of 60 people familiar with taste quality evaluation. The samples were provided in a random order using a random 3-digit number to ensure there was no bias during the evaluation.

[0134] IngredientsA16A14Isomalt49.5141%49.4922%Xylitol49.5141%49.4921%Lemon extract0.4500%0.4500%A160.0418%A140.0857%Citric acid0.2000%0.2000%Lemon flavor0.1500%0.1500%Mint flavor0.0800%0.0800%Carthamus Yellow0.0500%0.0500%Total100.0000%100.0000%

[0135] As a result of conducting a sensory evaluation on the manufactured hard candies, as shown in Table 15, it was confirmed that the preference for hard candies made using A16 of the present application was higher than that for hard candies using A14. In addition, in the case of the A14 sweetener, the high amount used resulted in a high intensity of off-flavor that impairs the unique refreshing taste of xylitol, and the sweetness appeal impairs the clean flavor of the candy, resulting in significantly lower overall preference. Accordingly, it was found that using A16 of the present application as a sweetener for manufacturing hard candies leads to excellent taste quality.

[0136] Category Sweetness Intensity Sweetness Attraction Intensity Lemon Mint Scent Intensity Ontitude Intensity Overall Preference A16A14A16A14A16A14A16A14A16A14A16A14Avg 5.785.884.244.973.233.723.623.855.355.02

[0137] 3-4. Chocolate Products

[0138] The chocolate was prepared as dark chocolate, and to produce sugar-free chocolate, isomalt, a disaccharide sugar alcohol, was used as a bulk sweetener. The above isomalt is suitable for the chocolate manufacturing process due to its low hygroscopicity, and since its sweetness is 0.5 times that of sugar and relatively low compared to other sugar alcohols (maltitol, sorbitol, xylitol), A16 or A14 was used at a concentration corresponding to 8 SEV by utilizing the sweetness evaluation results of the above-described example to compensate for the low sweetness of isomalt. The specific mixing ratio of the dark chocolate is shown in Table 16 below. First, isomalt and cocoa butter were added to the cocoa mass and thoroughly mixed with a mixer at 50°C for 10 minutes, and then passed through a 3-stage or 5-stage roller to finely reduce the average particle size to 15 to 20 µm. To the above-mentioned finely ground mixture, A16 or A14 was added along with lecithin, flavoring, and vanillin, and thoroughly mixed at 50°C for 60 minutes. After undergoing a tempering process, the mixture was poured into a mold and cooled to produce dark chocolate. A sensory evaluation was performed on the chocolate prepared as described above, and the evaluation was conducted on a panel of 60 people familiar with taste quality evaluation. The dark chocolate samples were provided in a random order using a random 3-digit number to ensure there was no bias during the evaluation.

[0139] IngredientsA16A14Isomalt38.820%38.820%Cacao mass47.110%47.110%Cacao butter13.4003%13.0746%A160.1197% A14 0.2454%Lecithin0.200%0.200%Vanilla flavor0.130%0.130%Milk flavor0.120%0.120%Vanillin0.100%0.100%Total100.0000%100.0000%

[0140] As a result of conducting a sensory evaluation on the manufactured dark chocolate, as shown in Table 17, the preference for the chocolate made using A16 was higher than that for the chocolate using A14. In the case of A14, it was confirmed that the overall preference was significantly lower because the strong aftertaste bitterness and sweetness attraction impair the unique flavor of the chocolate. Accordingly, it was confirmed that A16 has a weaker bitterness and less attraction compared to the A14 sweetener, thereby increasing the intensity of the cocoa flavor of the chocolate and resulting in excellent taste quality when applied to chocolate.

[0141] Category Sweetness Intensity Sweetness Attraction Intensity Bitterness Intensity Cocoa Flavor Intensity Overall Preference A16A14A16A14A16A14A16A14A16A14A16A14Avg4.804.823.524.584.235.623.893.555.955.52

[0142] 3-5. Chewing Gum Products

[0143] The chewing gum was prepared as sugar-free apple mint gum, and xylitol and isomalt, which are sugar alcohols, were used as bulk sweeteners for the production of the chewing gum. To supplement the sweetness of the sugar alcohols xylitol and isomalt, A16 or A14 was used at a concentration corresponding to 8 SEV, utilizing the sweetness evaluation results described above. The mixing ratio of the chewing gum is shown in Table 18 below. First, the gum base was placed in a Z-kneader and stirred while maintaining a temperature of 45 to 50°C until it became a dough-like consistency. Xylitol powder was added and mixed, and isomalt was added and stirred to ensure complete dispersion. Once the gum base and bulk sweeteners were evenly mixed, flavorings, emulsifiers, and sweeteners were added, and the mixture was stirred for about 10 minutes. After the mixing was completed, the dough was spread out on a sheeting roller, cut into stick gum shapes, and cooled to produce the chewing gum. A sensory evaluation was conducted on the chewing gum manufactured as described above, and the evaluation was conducted on a panel of 60 people familiar with taste quality evaluation. The chewing gum samples were provided in a random order using a random 3-digit number to ensure there was no bias during the evaluation.

[0144] Ingredients A16A14 flavor0.2100%0.2100%Total100.0000%100.0000%

[0145] As a result of conducting a sensory evaluation on the manufactured chewing gum, as shown in Table 19, the preference for chewing gum made using A16 was higher than that of the chewing gum sample using A14. In the case of A14, due to the characteristics of gum consumption, when chewing in the mouth, a bitter taste and off-flavor persisted after the refreshing sweetness of xylitol disappeared, and it was confirmed that the overall preference was significantly low. Accordingly, in the case of A16, the persistence of off-flavor or bitterness in the aftertaste was significantly low, and the sweetness combination with refreshing sugar alcohols such as xylitol was excellent. It was confirmed that the A16 sweetener exhibits excellent taste quality in various applications in the actual food industry.

[0146] Classification Sweetness Intensity Sweetness Attraction Intensity Bitterness Persistence Flavor Onset Intensity Overall Preference A16A14A16A14A16A14A16A14A16A14Avg 6.37 6.28 4.35 4.88 4.23 4.5 42.5 32.8 15.4 45.13

[0147] 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.

[0148]

[0149] [Consignment Number]

[0150] Name of depositing institution: Korean Culture Collection of Microorganisms

[0151] Trustee Number: KCCM13503P

[0152] Date of Trust: 20240805

[0153]

Claims

A food composition comprising a glucose-transferred steviol glycoside and a bulk sweetener, 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 food composition in which the glucose-transferred steviol glycoside comprises one or more glucose units added via an α-1,6 bond to a glucose unit connected to the 19th carbon of the steviol glycoside. In claim 1, A food 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, The above bulk sweetener is a food composition comprising sugar alcohols or sugars. In claim 3, The above bulk sweetener is xylitol and / or isomalt, food composition. In claim 1, The above composition is a food composition that is a sweetener composition for food. A food comprising the food composition of any one of claims 1 to 5. In claim 6, The above food is at least one selected from the group consisting of beverages, confectionery, baked goods, desserts, and candies. In claim 6, The above food is at least one selected from the group consisting of coffee beverages, yogurt beverages, soy milk beverages, cookies, chocolates, candies, and chewing gum. In claim 6, A food having a content of 0.0001 to 3 parts by weight of the glucose-transferred steviol glycoside based on 100 parts by weight of the above food.

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