Composition comprising glucosylated steviol glycoside

Abstract

The present application relates to a composition comprising a glucosylated steviol glycoside, and a sweetener comprising same, the composition being excellent at improving sweetness. The glucosylated steviol glycoside is mixed with at least one selected from the group consisting of surgars and sugar alcohols, and thus, the powder of such a mixture is suppressed or prevented from clumping, and the sweetener in powder form can be stored at room temperature for a long time.

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. 10-2025-0012717 filed on January 31, 2025, Korean Patent Application No. 10-2025-0119488 filed on August 26, 2025, and Korean Patent Application No. 10-2025-0212677 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 sweetener 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, possess a sweetness approximately 200 to 400 times greater than sugar depending on the specific component, and in particular, rebaudioside A, rebaudioside D, and rebaudioside M are primarily used as texture-enhancing sweeteners. 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] In addition, sweeteners and seasonings are generally manufactured in powder form. Consequently, the solidification phenomenon, a common issue that occurs during the distribution of powder products, is also a problem for sugar. Solidification is a phenomenon in which the particles of the contents of a powder product absorb surrounding moisture and clump together during distribution; the smaller the powder particles, the larger the surface area for moisture absorption, and the more severe the phenomenon becomes. Since this solidification phenomenon degrades product quality and causes inconvenience to consumers, methods have been proposed to prevent it by adding silicon dioxide, pectin, starch, etc., as anti-solidification agents to the sweetener powder and mixing them. However, since these anti-solidification agents are added in small quantities when manufacturing the sweetener composition, it is difficult to distribute them evenly throughout the sweetener composition using this previously known simple mixing method, resulting in a problem where the solidification phenomenon of the powder cannot be effectively suppressed.

[0010] There is a need for measures to solve the conventional problems regarding such steviol glycosides and sweeteners.

[0011]

[0012] [Prior Art Literature]

[0013] U.S. Patent Publication US 12053005 B2

[0014] Korean Registered Patent Publication KR 10-2421798

[0015] U.S. Patent Publication US 2024-0090551 A1

[0016]

[0017] The present application aims to provide a composition and a sweetener comprising a glucose-transferred stevia glycoside in order to improve off-flavors and prevent solidification of the sweetener by utilizing the glucose-transferred stevia glycoside.

[0018] In addition, the present application aims to provide a method for inhibiting or preventing solidification of a mixture of at least one selected from the group consisting of sugars and sugar alcohols and a glucose-transferred steviol glycoside.

[0019]

[0020] One aspect of the present application provides a composition comprising at least one selected from the group consisting of sugars and sugar alcohols and a glucose-transferred steviol glycoside, 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 wherein the glucose-transferred steviol glycoside has one or more glucose units added to the glucose connected to the 19th carbon of the steviol glycoside via an α-1,6 bond.

[0021] Another aspect of the present application provides a sweetener comprising the above composition.

[0022] Additionally, another aspect of the present application provides a method for inhibiting or preventing solidification of a mixture of at least one selected from the group consisting of sugars and sugar alcohols and a glucose-transferred steviol glycoside, comprising the step of mixing at least one selected from the group consisting of sugars and sugar alcohols with a glucose-transferred steviol glycoside, 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 wherein the glucose-transferred steviol glycoside has one or more glucose units added to a glucose connected to the 19th carbon of the steviol glycoside via an α-1,6 bond.

[0023]

[0024] The present application will be described in detail below.

[0025]

[0026] 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.

[0027] [Chemical Formula 1]

[0028]

[0029] 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.

[0030] 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.

[0031] 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.

[0032] 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.

[0033] 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.

[0034] 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.

[0035] 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.

[0036]

[0037] One aspect of the present application provides a composition comprising at least one selected from the group consisting of sugars and sugar alcohols and the glucose-transferred stevia glycoside.

[0038] The content of the glucose-transferred steviol glycoside in the composition of the present application may be 0.001 to 3 parts by weight based on 100 parts by weight of the composition, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 0.001, 0.005, 0.007, 0.01, 0.05, 0.07, 0.1, 0.2, 0.25, and 0.3 parts by weight, and an upper limit selected from the group consisting of 0.3, 0.4, 0.5, 0.7, 1, 1.5, 2, 2.5, and 3 parts by weight; for example, the content of the glucose-transferred steviol glycoside may be 0.001 to 2.5 parts by weight, 0.005 to 2 parts by weight, 0.007 to 1.5 parts by weight, or 0.01 parts by weight based on 100 parts by weight of the composition. It may be up to 1 part by weight, 0.05 to 0.7 parts by weight, 0.07 to 0.5 parts by weight, 0.1 to 0.4 parts by weight, 0.2 to 0.4 parts by weight, 0.25 to 0.3 parts by weight, or 0.3 to 0.4 parts by weight.

[0039] The above sugars may be at least one of monosaccharides, disaccharides, and oligosaccharides.

[0040] 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, lactose, maltose, trehalose, turanose, or cellobiose, but are not limited thereto.

[0041] The above sugar alcohols are substances formed by reducing the carbonyl group of sugars and may also be called sugar alcohols, and may be, for example, erythritol, xylitol, arabitol, mannitol, sorbitol, maltitol, or lactitol, but are not limited thereto.

[0042] At least one selected from the group consisting of the above sugars and sugar alcohols may be at least one selected from the group consisting of bulk sweeteners, rare sugars, nutritional sweeteners, non-nutritional sweeteners, non-sugar sweeteners, and high-intensity sweeteners.

[0043] The content of at least one selected from the group consisting of sugars and sugar alcohols in the composition of the present application may be 90 to 99.99 parts by weight based on 100 parts by weight of the composition, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 90, 91, 92, 93, 94, 95, 96, 97, 98, and 99 parts by weight, and an upper limit selected from the group consisting of 99.7, 99.75, 99.8, 99.85, 99.9, 99.95, and 99.99 parts by weight; for example, the content of at least one selected from the group consisting of sugars and sugar alcohols may be 91 to 99.99 parts by weight, 92 to 99.95 parts by weight, 93 to 99.9 parts by weight, and 94 to It may be 99.9 parts by weight, 95 to 99.85 parts by weight, 96 to 99.8 parts by weight, 97 to 99.8 parts by weight, 98 to 99.75 parts by weight, or 99 to 99.75 parts by weight.

[0044] In the above composition, the content ratio of at least one selected from the group consisting of the glucose-transferred steviol glycoside, sugars, and sugar alcohols may be 0.001:99.99 to 3:90, for example, 0.02:99.9 to 0.5:99 or 0.3:99.7 to 0.5 to 99.5, but is not limited thereto.

[0045] The above bulk sweetener may include sugar alcohols or sugars, and for example, the above 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, xylulose, psicose, turanos, cellobiose, glucosamine, mannosamin, fucose, fuculose, glucuronic acid, gluconic acid, gluconolactone, abequeose, galactosamine, It may include at least one selected from the group consisting of xylooligosaccharides (xyllotrios, 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.

[0046] The above sugars may include rare sugars, and the rare sugars may include at least one selected from the group consisting of D-allose, D-psicose (D-allulose), L-ribose, D-tagatose, L-glucose, L-fucose, L-arabinose, D-turanose, and D-leucrose.

[0047] The above nutritional sweetener may include at least one selected from the group consisting of D-psychose, fructose, glucose, mannose, arabinose, galactose, xylose, rhamnose, ribose, fucose, sucrose, maltose, lactose, maltitol, xylitol, erythritol, sorbitol, mannitol, palatinite, maltotriitol, maltotetraitol, and D-tagatose.

[0048] The above non-nutritive sweetener may include at least one selected from the group consisting of aspartame, acesulfame salts, saccharins, cyclamates, sucralose, alitame, neotame, steviosides, glycyrrhizin, Lo Han Guo, neohesperidin dihydro chalcone, monatin, monellin, thaumatin, and brazzein.

[0049] The above-mentioned non-carbohydrate sweeteners are rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, dulcoside A, dulcoside B, rubusoside, stevia, stevioside, mogroside IV, mogroside V, Luohanguo sweetener, siamenoside, monatin and its salts (monatin SS, RR, RS, SR), curculin, glycyrrhizic acid and its salts, taumatin, monellin, marvinlin, bracein, hernandulcin, phyllodulcin, glycyphyllin, phloridzin, trilovatin, bayunosid, osladin, polypodoside A, pterocarioside A, pterocarioside B, mucurozioside, flomisoside I, periandrin I, abrusoside A, and It may include at least one selected from the group consisting of cyclocarioside I.

[0050] The above-mentioned high-potency sweetener (HPS) may include modified HPS. The modified HPS may include naturally modified HPS. For example, modified HPS may include fermented, enzyme-contacted, or induced or substituted HPS, but is not limited thereto. Specifically, one or more modified HPS may be used in a form combined with one or more HPS, or one or more modified HPS may be used without HPS. Thus, modified HPS may be used instead of HPS or in combination with HPS. However, for the sake of brevity, the description of the embodiments of this application does not explicitly present modified HPS as an alternative to unmodified HPS, but modified HPS may be used instead of HPS.

[0051] In one embodiment, the sugar may be allulose, and the sugar alcohol may be erythritol.

[0052] The above allulose (D-allulose) can be called D-psicose or D-ribo-2-hexulose; it is a monosaccharide containing six carbon atoms, a ketose containing a ketone group, and has the chemical formula C6H12O6. Allulose is a low-energy monosaccharide that exists in small amounts in natural products and is mainly used in zero-calorie diet drinks. The sweetness of allulose is about 70% of the sweetness of sugar.

[0053] The above erythritol is a natural sugar alcohol found in fruits and fermented foods, produced by fermenting glucose. The above erythritol is a sweetener that does not cause tooth decay and is also used as a diet sweetener because it has almost no calories. The sweetness of the above erythritol is about 70% of that of sugar.

[0054] In one embodiment, the composition of the present application may include the glucose-transferred steviol glycoside and allulose.

[0055] In one embodiment, the composition of the present application may include the glucose-transferred steviol glycoside and erythritol.

[0056] In addition, the above composition may be used in the form of a liquid composition, a solid composition, or a gelling composition, and may be used in the form of a powder. The above composition may be a powder composition.

[0057] The composition of the present application can inhibit or prevent the solidification of the powder.

[0058] The aforementioned solidification is a phenomenon in which the particles of the contents of a powder product absorb surrounding moisture and clump together. This can occur during the flow and storage of the powder product, and the smaller the size of the powder particles, the larger the surface area for moisture absorption, which exacerbates the phenomenon. When tabletop sweeteners are distributed, moisture absorption may occur due to various factors, causing the substances to form larger shapes and change into a hard state. Since a higher amount of solidification in the tabletop sweetener product implies reduced storage stability and significantly lower marketability, resulting in lower quality, managing such solidification is very important.

[0059] In a specific embodiment of the present application, a tabletop powdered sweetener was prepared using a composition comprising the glucose-transferred steviol glycoside of the present application and erythritol or allulose, water was added to induce solidification, and after being stored at room temperature for at least one week, the solidification rate (%) of the tabletop powdered sweetener was investigated. As a result, it was confirmed that when the composition of the present application was used, the solidification rate was low, allowing the quality to be maintained for a long period at room temperature, thus demonstrating excellent storage stability.

[0060] In this application, the term "inhibition or prevention of solidification" means that, based on the fine amount and solidification amount of the powder, when the solidification rate is calculated by measuring the ratio of the solidification amount to the fine amount, the solidification rate is lower than that of a powder containing an α-1,4 glucose-transferred steviol glycoside used as a conventional sweetener. The solidification rate is a percentage representing the ratio of such solidified and clumped solidified material to the fine amount. The inhibition or prevention of solidification may mean that the solidification rate of the powder containing the composition of this application is lower by at least 1% to a maximum of 4% compared to a powder containing an α-1,4 glucose-transferred steviol glycoside used as a conventional sweetener, and if it is lower by 1% or more, it may be considered as inhibition or prevention of solidification, but is not limited thereto.

[0061] The above composition may be used for the manufacture of sweetener products, and may also be used in the form of a semi-finished product or a syrup or powder during the process.

[0062] The composition of the present application may be used as a sweetener, and said composition may be a sweetener composition or a composition for use in a sweetener, may be added to a sweetener or used together with other sweetener components, and may be used appropriately according to conventional methods.

[0063] Accordingly, the composition of the present application may be a sweetener composition in which solidification is inhibited or prevented, and specifically, may be a composition for inhibiting or preventing solidification of a sweetener, and may be a composition for use in inhibiting or preventing solidification of a sweetener.

[0064] Additionally, the composition of the present application may further include at least one additive selected from the group consisting of nutrients, flavoring agents, natural carbohydrates, dietary fiber, 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 acidifier may be an organic acid, and the acidifier may further include at least one additive 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 such additives may be selected in the range of 0.01 to 1 part by weight based on 100 parts by weight of the sweetener of the present application, specifically, 0.05 to 0.2 parts by weight, and particularly 0.1 part by weight, but is not limited thereto.

[0065] The composition of the present application comprises at least one selected from the group consisting of glucose-transferred steviol glycosides, sugars, and sugar alcohols, thereby improving the inherent bitterness of the glucose-transferred steviol glycosides and enhancing the sweetness preference. The composition has excellent sweetness and low off-flavor or odor. When a sweetener is manufactured using the composition, the off-flavor or odor of the steviol glycoside sweetener is improved, resulting in excellent taste quality.

[0066] In addition, the glucose-transferred steviol glycoside of the present application has a superior effect of improving sweetness compared to widely used α-1,4-linked glucose-transferred steviol glycosides, exhibits a more uniform sweetener distribution when mixed, has excellent processability with less stickiness after coating, and can suppress or prevent the solidification of the powder.

[0067]

[0068] Another aspect of the present application provides a sweetener comprising the above composition.

[0069] The sweeteners of the present application include, but are not limited to, general sweeteners, health sweeteners, and sweeteners for medical (or, patient) use.

[0070] In the sweetener of the present application, the content of the glucose-transferred steviol glycoside may be 0.001 to 3 parts by weight based on 100 parts by weight of the sweetener, and may be included in a range selected from the group consisting of a lower limit selected from the group consisting of 0.001, 0.005, 0.007, 0.01, 0.05, 0.07, 0.1, 0.2, 0.25, and 0.3 parts by weight, and an upper limit selected from the group consisting of 0.3, 0.4, 0.5, 0.7, 1, 1.5, 2, 2.5, and 3 parts by weight; for example, the content of the glucose-transferred steviol glycoside may be 0.001 to 2.5 parts by weight, 0.005 to 2 parts by weight, or 0.007 to 1.5 parts by weight based on 100 parts by weight of the composition. It may be 0.01 to 1 part by weight, 0.05 to 0.7 parts by weight, 0.07 to 0.5 parts by weight, 0.1 to 0.4 parts by weight, 0.2 to 0.4 parts by weight, 0.25 to 0.3 parts by weight, or 0.3 to 0.4 parts by weight. In the sweetener of the present application, the content of at least one selected from the group consisting of sugars and sugar alcohols may be 90 to 99.99 parts by weight based on 100 parts by weight of the sweetener, and a lower limit of any one selected from the group consisting of 90, 91, 92, 93, 94, 95, 96, 97, 98, and 99 parts by weight, and in the group consisting of 99.7, 99.75, 99.8, 99.85, 99.9, 99.95, and 99.99 parts by weight It may be included in an amount within a range selected from a group consisting of any one upper limit selected, for example, at least one amount selected from the group consisting of sugars and sugar alcohols may be 91 to 99.99 parts by weight, 92 to 99.95 parts by weight, 93 to 99.9 parts by weight, 94 to 99.9 parts by weight, 95 to 99.85 parts by weight, 96 to 99.8 parts by weight, 97 to 99.8 parts by weight, 98 to 99.75 parts by weight, or 99 to 99.75 parts by weight, based on 100 parts by weight of the composition.In the above sweetener, the content ratio of at least one selected from the group consisting of the glucose-transferred steviol glycoside, sugars, and sugar alcohols may be 0.001:99.99 to 3:90, for example, 0.02:99.9 to 0.5:99 or 0.3:99.7 to 0.5 to 99.5, but is not limited thereto.

[0071] In one embodiment, the sweetener of the present application may be a tabletop sweetener.

[0072] In one embodiment, the sweetener of the present application may be a powdered sweetener.

[0073] In one embodiment, the sweetener of the present application may be a tabletop powder sweetener.

[0074] The sweetener of the present application may be in powder form, and solidification of the powder may be inhibited or prevented by including the composition.

[0075] In one embodiment, the sweetener of the present application may be a sweetener in which solidification is inhibited or prevented.

[0076]

[0077] Another aspect of the present application provides a method for inhibiting or preventing solidification of a mixture of at least one selected from the group consisting of sugars and sugar alcohols and a glucose-transferred steviol glycoside.

[0078] The above method for inhibiting or preventing solidification includes the step of mixing at least one selected from the group consisting of sugars and sugar alcohols with a glucose-transferred steviol glycoside.

[0079] The above mixing step may involve homogenizing at least one selected from the group consisting of sugars and sugar alcohols with a glucose-transferred steviol glycoside. The homogenization may mean dispersing each component of a heterogeneous mixture into fine particles or molecules to make the whole homogeneous. The homogenization may be performed using means or equipment that a person skilled in the art can adopt for homogenization, for example, using a high-speed agitator, a colloid mill, an ultrasonicator, etc.

[0080] The above mixing step may involve mixing such that the content of the glucose-transferred steviol glycoside is 0.001 to 3 parts by weight based on 100 parts by weight of the mixture.

[0081] In addition, the mixing step may involve mixing such that the content of at least one selected from the group consisting of sugars and sugar alcohols is 90 to 99.99 parts by weight based on 100 parts by weight of the mixture.

[0082] In addition, in the mixing step, at least one mixing ratio selected from the group consisting of the glucose-transferred steviol glycoside, sugars, and sugar alcohols may be 0.001:99.99 to 3:90, and for example, the mixing ratio may be 0.02:99.9 to 0.5:99 or 0.3:99.7 to 0.5 to 99.7, but is not limited thereto.

[0083]

[0084] The glucose-transferred steviol glycoside of the present application has improved bitterness and sweetness palatability, and has excellent sweetness quality with minimal off-flavor or off-odor. When a sweetener is manufactured using a composition containing such a glucose-transferred steviol glycoside, the off-flavor or off-odor of the steviol glycoside sweetener is improved, resulting in excellent taste quality.

[0085] In addition, compared to widely used α-1,4-linked glucose-transferred steviol glycosides, the glucose-transferred steviol glycoside of the present application has a superior effect of improving sweetness when combined with a number of non-nutritive carbohydrates, exhibits a more uniform sweetener distribution when mixed, and has excellent processability with less stickiness after coating.

[0086] In particular, when at least one selected from the group consisting of glucose-transferred steviol glycosides, sugars, and sugar alcohols is mixed and used as in the composition of the present application, solidification of the powder of the mixture can be suppressed or prevented, so the sweetener in powder form can be stored at room temperature for a long time, thereby extending the shelf life and having an advantage in maintaining quality.

[0087] 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 description below.

[0088]

[0089] Figure 1 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside A by the Lactobacillus mali CJST242 strain.

[0090] Figure 2 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside C by the Lactobacillus mali CJST242 strain.

[0091] Figure 3 is an HPLC chromatogram of the result of the glycosylation reaction of ribaudioside F by the Lactobacillus mali CJST242 strain.

[0092] Figure 4 is an HPLC chromatogram of the result of the glycosylation reaction of stevioside by the Lactobacillus mali CJST242 strain.

[0093] Figure 5 is an HPLC chromatogram of the result of the glycosylation reaction of dulcosides by the Lactobacillus mali CJST242 strain.

[0094] Figure 6 is an HPLC chromatogram of the glycosylation reaction results of rubusoside by the Lactobacillus mali CJST242 strain.

[0095] 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.

[0096] Figure 8 shows the results of comparing the solidification rates of the tabletop sweetener on the left containing the glucose-transferred steviol glycoside of the present application and the tabletop sweetener on the right containing A14.

[0097]

[0098] The present application will be explained in more detail below through examples.

[0099] 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.

[0100]

[0101] Preparation Example 1: Preparation of glucose-transferred steviol glycosides

[0102] The preparation of glucose-transferred steviol glycosides was carried out according to the method described in Korean Patent Publication No. 10-2021-0114899.

[0103] 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.

[0104] 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.

[0105] 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.

[0106] 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).

[0107] The results are listed in Tables 1 to 4 below.

[0108] 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").

[0109] 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").

[0110] 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.

[0111] 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.

[0112] 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.

[0113] w Weak signals.

[0114] * The chemical shift of each signal may not be exact value due to signal overlapping

[0115] a,b,c,d,e,f,g,hThe assignment of signals at same alphabet column may be exchangeable.

[0116]

[0117] 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.

[0118] 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.

[0119] 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.

[0120] 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.

[0121]

[0122] Example 1: Setting Equivalent Sweetness of Glucose-Transferred Steviol Glycosides

[0123] 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.

[0124] 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.

[0125] Sweetness evaluation of Reb M

[0126] 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.

[0127] 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

[0128] 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.

[0129] 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

[0130] 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.

[0131]

[0132] Example 2: Setting Equivalent Sweetness for α-1,4 Glucose-Transferred Steviol Glycosides

[0133] 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.

[0134] 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

[0135] 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.

[0136]

[0137] Example 3: Sensory evaluation of a sweetener prepared using a glucose-transferred steviol glycoside

[0138] 3-1. Combination of glucose-transferred steviol glycoside and allulose of the present application

[0139] In order to compare and analyze the sweetness quality of the sweetener prepared using A16 of the present application in Preparation Example 1 above, a sensory evaluation was performed. A sweetener was prepared by mixing A16 and allulose, and to compare taste quality, a sweetener prepared by mixing allulose with an alpha-1,4 linked glucose-transferred steviol glycoside (hereinafter A14), which is widely used as a conventional sweetener, was used as a comparison group. The input concentrations of A16 and A14 were determined by applying the sweetness values ​​of 66.8 times and 32.6 times, respectively, calculated in Examples 1 and 2 above. During the sensory evaluation, the sweetness of the aqueous solution containing the sweetener was provided as 8 SEV.

[0140] Specifically, to prepare the composition, the raw materials were accurately weighed according to the content ratios shown in Table 8 below; however, the formulation of the composition is not limited to the content ratios in Table 8 below and may encompass a broader range. Next, the weighed allulose was spread onto a wide, flat stainless steel dish, and then a small amount of the weighed A16 and A14 were each evenly sprinkled over the already spread allulose powder. Subsequently, purified water was uniformly sprayed over the entire spread area using a spray bottle and mixed to ensure even mixing. The mixed sweetener was dried in an oven at a temperature of 60 to 150°C to ensure sufficient removal of moisture. Meanwhile, this drying process can also be performed using a fluid dryer and is not limited to the method presented in this application.

[0141] IngrtdientsA16A14Allulose99.7000%99.5000%A160.3000%A14 0.5000%Total100.0000%100.0000%

[0142] Subsequently, to prepare a powdered sweetener aqueous solution, purified water was mixed with 6.717% of the sweetener containing A16, and purified water was mixed with 6.728% of the sweetener containing A14 to prepare a mixed solution. The aqueous solution was divided into 10 mL portions in tasting containers and provided for sensory evaluation. The sensory evaluation was conducted with a panel of 100 individuals familiar with evaluating taste quality. Samples were provided using a random 3-digit number to ensure there was no bias during evaluation. As a result, as shown in Table 9 below, it was confirmed that the sweetener aqueous solution containing A16 showed higher preference compared to the sweetener aqueous solution containing A14. Accordingly, it was confirmed that A16 exhibits superior taste quality in actual application cases in the field of sweeteners.

[0143] Classification Sweetness Intensity Flavor Intensity Overall Preference A16A14A16A14A16A14Avg 5.945.822.652.945.605.22

[0144] *bolded numbermeans the result of a significant difference (p<0.05) for each evaluation.

[0145] 3-2. Combination of glucose-transferred steviol glycoside and erythritol of the present application

[0146] A sensory evaluation was performed to compare and analyze the sweetness of the sweeteners prepared using the above A16. The sensory evaluation was conducted in the same manner as in Example 3-1, and erythritol was used in combination instead of allulose. In addition, each ingredient was accurately weighed according to the mixing ratios shown in Table 10 below.

[0147] Ingredients A16 A14 Erithritol 99.75 00% 99.5 700% A16 0.25 00% A14 0.4 300% Total 100.00 00% 100.00 00%

[0148] Subsequently, to prepare a powdered sweetener aqueous solution, purified water was mixed with a sweetener containing 5.717% of the above A16, and purified water was mixed with a sweetener containing 5.738% of the above A14 to prepare a mixed solution. As a result, as shown in Table 11 below, it was confirmed that the sweetener aqueous solution containing A16 had a higher palatability compared to the sweetener aqueous solution containing A14. Accordingly, it was confirmed that A16 exhibits excellent taste quality in actual sweetener applications.

[0149] Classification Sweetness Intensity Flavor Intensity Mouthfeel Intensity Overall Preference A16A14A16A14A16A14A16A14Avg6.246.062.462.813.763.535.565.18

[0150] *bolded numbermeans the result of a significant difference (p<0.05) for each evaluation.

[0151]

[0152] Example 4: Confirmation of improved solidification of sweetener prepared using glucose-transferred steviol glycosides

[0153] Since tabletop sweeteners are stored at room temperature for extended periods and utilized as sweeteners, it is important that the powder itself does not solidify and maintains its powdered state for a long time. Accordingly, the solidification rate of the powdered sweetener prepared using the composition of the present application was evaluated.

[0154] The solidification rate of a tabletop sweetener containing the glucose-transferred steviol glycoside of the present application during long-term storage was evaluated. First, a tabletop sweetener was prepared by mixing 0.3% of the glucose-transferred steviol glycoside (A16) and 99.7% of erythritol or allulose, and then water was added up to 1% to induce forced solidification of the tabletop sweetener. Afterward, solidification was allowed to proceed by storing at room temperature for one week, and then the fine amount (g) and solidified amount (g) of the tabletop sweetener were measured to calculate the solidification rate (%). The solidification rate represents the percentage value of the solidified amount relative to the total amount of the tabletop sweetener. The solidification rate measurement experiment was conducted five times to derive an average value. At this time, a tabletop sweetener containing the above A14 and erythritol or allulose was prepared and used as a comparison group. The experimental results regarding the solidification rate of the tabletop sweetener containing the above-mentioned erythritol are shown in Table 12 below, and the experimental results regarding the solidification rate of the tabletop sweetener containing the above-mentioned allulose are shown in Table 13 below.

[0155] Classification 1st Round 2nd Round 3rd Round 4th Round 5th Round Average A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 Amount of rice (g) 25 1.72 23 2.85 47.97 47.05 50.78 47.94 3.72 46.46 50.58 49.99 88.95 48 4.85 Amount of solidified rice (g) 82.98 93.31 2.23 13.45 10.54 12.33 14.87 13. Total (g) 334.73 26.26 0.26 0.56 1.32 60.23 58.59 59.67 62.47 63.51 115.45 61 14.022 Solidification Rate 24.79% 28.60% 20.32% 22.23% 17.19% 20.47% 25.38% 22.14% 19.03% 21.29% 22.95% 25.58%

[0156] Category 1st Round 2nd Round 3rd Round 4th Round 5th Round Average A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 A16 A14 Amount of Rice (g) 205.22 200.9 141.7 240.8 636.8 735.6 144.7 944.3 739.2 538.6 173.5 772.07 2 Bread Amount (g) 94.5 8100.4 918.7 820.5 421.7 324.19 1 5.6 11 6.8 32 4.4 5 22.5 9 35.0 33 6.9 28 Total (g) 29 9.8 30 0.4 6 0.5 6 1.4 5 8.6 5 9.8 6 0.4 6 1.2 6 3.7 6 1.2 10 8.6 10 8.8 Solidification Rate 31.55% 33.45% 31.04% 33.45% 37.08% 40.45% 25.84% 27.50% 38.38% 36.91% 32.26% 33.94%

[0157] As a result, as shown in Tables 12 to 13 and Figure 8, the tabletop sweetener containing A16 of the present application and bulk sweeteners such as erythritol or allulose exhibited an average lower solidification rate compared to the case where A14 was used, confirming that the glucose-transferred steviol glycoside of the present application has excellent performance as a tabletop sweetener when combined with bulk sweeteners, and can be dissolved in food with consistent quality even during long-term storage.

[0158]

[0159] Preparation Example 2: Formulation of a sweetener

[0160] A sweetener comprising the above A16, allulose, and Rebaudioside M was prepared. The prepared sweetener has a formulation suitable for use as a cooking ingredient or for immediate consumption by adding it to water, other liquids, or other foods. Additionally, the sweetener may be packaged in bulk or in individual single-serving packages. A trace amount of flavoring (e.g., 0.01g or less) or dietary fiber may be further added to the sweetener. The addition of dietary fiber can result in a better mouthfeel and improved sweetness.

[0161]

[0162] 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.

[0163]

[0164] [Consignment Number]

[0165] Name of depositing institution: Korean Culture Collection of Microorganisms

[0166] Trustee Number: KCCM13503P

[0167] Date of Trust: 20240805

[0168]

Claims

A composition comprising at least one selected from the group consisting of sugars and sugar alcohols and a glucose-transferred steviol glycoside, 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 in which the above sugars are at least one of monosaccharides, disaccharides, and oligosaccharides. In claim 1, A composition in which the above sugar is allulose. In claim 1, A composition in which the above sugar alcohols are erythritol. In claim 1, A composition comprising 0.001 to 3 parts by weight of the glucose-transferred steviol glycoside based on 100 parts by weight of the above composition. In claim 1, A composition comprising, based on 100 parts by weight of the above composition, at least one selected from the group consisting of sugars and sugar alcohols in an amount of 90 to 99.99 parts by weight. In claim 1, A composition in which the ratio of at least one selected from the group consisting of the glucose-transferred steviol glycoside and the sugars and sugar alcohols is 0.001:99.99 to 3:

90. In claim 1, The above composition is a powder. In claim 8, The above composition is a composition that inhibits or prevents the solidification of the above powder. A sweetener comprising the composition of any one of claims 1 to 10. In claim 11, The above sweetener is a sweetener in which solidification is inhibited or prevented. A method for inhibiting or preventing solidification of a mixture of at least one selected from the group consisting of sugars and sugar alcohols and a glucose-transferred steviol glycoside, comprising the step of mixing at least one selected from the group consisting of sugars and sugar alcohols with a glucose-transferred steviol glycoside. 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 method for inhibiting or preventing solidification, wherein the glucose-transferred steviol glycoside is one in which one or more glucose units are added to the glucose connected to the 19th carbon of the steviol glycoside via an α-1,6 bond.

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