阅读说明：本技术 包含转果糖基化的甜菊糖苷的组合物 (Compositions comprising transfructosylated steviol glycosides ) 是由 金贞银 杨泰周 朴晟喜 姜仁声 金成俌 秋善 于 2020-04-09 设计创作，主要内容包括：本申请涉及一种包含转果糖基化的甜菊糖苷的甜味剂组合物以及一种改善甜菊糖苷甜味的方法,所述方法包括将甜菊糖苷转化为转果糖基化的甜菊糖苷。(The present application relates to a sweetener composition comprising a transfructosylated steviol glycoside and a method of improving the sweetness of a steviol glycoside, which method comprises converting the steviol glycoside into a transfructosylated steviol glycoside.)

1. A sweetener composition comprising a transfructosylated steviol glycoside wherein the fructose is linked to the 19-OH site of the steviol glycoside or to a glucose conjugated thereto.

2. A sweetener composition according to claim 1 where the transfructosylated steviol glycoside is a form in which fructose is linked to the steviol glycoside through a β - (2,6) bond.

3. A sweetener composition according to claim 1 where the steviol glycoside is stevioside or rebaudioside A.

4. A sweetener composition according to claim 1 where the transfructosylated steviol glycoside is in a form where 1-3 fructose molecules are attached to the steviol glycoside.

5. A sweetener composition according to claim 1 where the transfructosylated stevioside has an improved bitter taste compared to the stevioside.

6. A sweetener composition according to claim 1 where the transfructosylated stevioside has an improved sweetness compared to the stevioside.

7. A food composition comprising the composition according to any one of claims 1 to 6.

8. A method for improving sweetness of stevioside comprises converting stevioside into transfructosylated stevioside,

wherein the transfructosylated steviol glycoside is in a form in which fructose is linked to the 19-OH site of the steviol glycoside or to glucose conjugated thereto.

9. The method of claim 8, wherein the transfructosylated steviol glycoside is in a form in which fructose is linked to the steviol glycoside via a β - (2,6) bond.

10. The method of claim 8, wherein the steviol glycoside is stevioside or rebaudioside A.

11. The method of claim 8, wherein the transfructosylated steviol glycoside is in a form in which 1-3 fructose molecules are attached to the steviol glycoside.

Technical Field

The present disclosure relates to sweetener compositions comprising transfructosylated steviol glycosides.

Background

With the World Health Organization (WHO) recommending a reduction in daily sugar intake due to fear of diseases (obesity) caused by sugar intake, governments of developed countries are actively discussing various policies aimed at reducing sugar intake. Therefore, as the market for developing various alternative sweeteners increases, alternative sweeteners are continually being developed and commercialized. As alternative sweeteners, these are the subject of various variations in the form of synthetic high intensity sweeteners (e.g., saccharin, aspartame, sucralose, etc.), synthetic sugar alcohols (e.g., maltitol and xylitol), and high intensity sweeteners (e.g., rebaudioside a and licorice). Despite this, consumer demand for natural sweeteners has steadily increased due to concerns about the safety of synthetic sweeteners; however, natural sweeteners do not completely replace existing low-calorie and zero-calorie products based on synthetic sweeteners due to limitations in the flavor characteristics (i.e., unpleasant odor and off-taste) that are characteristic of natural sweeteners.

One natural high intensity sweetener that has received much attention in recent years is Stevia sugar extracted from the leaves of Stevia rebaudiana Bertoni. Stevioside is a natural substance, has sweetness 200 to 300 times that of sugar, and consists of stevioside, rebaudioside a and the like. Although stevioside and rebaudioside a show high sweetness, the bitterness is strong, which limits use.

Methods for improving the sweetness of stevioside and rebaudioside A include methods of transferring sugars using enzymes. As a method for transferring saccharides using an enzyme, a method for transferring 1 to 12 glucose molecules to steviol glycoside using CGTase is widely used in the art (Korean patent application No. 10-1991-0020769). However, this method has a disadvantage in that all glucose molecules transferred to steviol glycosides are degraded by intestinal microbes, thereby increasing calories. Therefore, there is a need for a steviol glycoside in which a saccharide other than glucose is transferred.

Disclosure of Invention

[ problem ] to

In this case, the present inventors completed the present disclosure by confirming that transfructosylated steviol glycosides have improved bitterness compared to steviol glycosides and show superior sweetness preference and overall preference.

[ solution ]

It is an object of the present disclosure to provide a sweetener composition comprising a transfructosylated steviol glycoside.

It is another object of the present disclosure to provide a food composition comprising the sweetener composition.

It is still another object of the present invention to provide a method for improving sweetness of steviol glycosides, comprising converting steviol glycosides into transfructosylated steviol glycosides.

[ advantageous effects ]

The sweetener compositions of the present disclosure comprise transfructosylated steviol glycosides, thereby having improved bitterness and exhibiting superior sweetness preference and overall preference as compared to steviol glycosides, and thus can be used as high intensity sweeteners.

Drawings

FIG. 1 shows sensory descriptive analysis results of transfructosylated rebaudioside A (RA-Fru) and Rebaudioside A (RA).

Figure 2 shows the results of sensory descriptive analysis of transfructosylated steviol glycosides (STV-Fru) and steviol glycosides (STV).

Detailed Description

Hereinafter, the present disclosure will be described in detail. Meanwhile, each of the explanations and exemplary embodiments disclosed herein may be applied to other explanations and exemplary embodiments. That is, all combinations of the various factors disclosed herein are within the scope of the present disclosure. Furthermore, the scope of the present disclosure should not be limited by the specific disclosure provided below. Meanwhile, the terms defined herein may be equally applied to other aspects of the present disclosure.

To achieve the objects of the present disclosure, one aspect of the present disclosure may be to provide transfructosylated steviol glycosides or sweetener compositions comprising transfructosylated steviol glycosides. Transfructosylated steviol glycosides can be produced by the above-described process.

As used herein, the term "steviol glycoside" refers to a natural sweetener and may be tabulated by chemical formula 1 below.

[ chemical formula 1]

In chemical formula 1, at R1, hydrogen (H) may be bound, or 1 to 3 glucose molecules may be bound through β -bonds; at R₂Here, 1 molecule of glucose, xylose or rhamnose may be bound through a β -bond, and 0 to 2 glucose molecules may be bound through a β -bond, but is not limited thereto.

The steviol glycoside may be one or more selected from stevioside, rubusoside, dulcoside a, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F and rebaudioside M, and specifically may be stevioside or rebaudioside a, but is not limited thereto.

Specifically, the steviol glycoside may be stevioside represented by the following chemical formula 2 or rebaudioside a represented by the following chemical formula 3.

[ chemical formula 2 ]

[ chemical formula 3]

As used herein, the term "transfructosylated steviol glycoside" may refer to a compound in which fructose is linked (transferred) to the steviol glycoside.

In particular, the transfructosylated steviol glycoside may be in a form in which fructose is linked to the 19-OH site of the steviol glycoside or to glucose conjugated thereto.

In particular, the transfructosylated steviol glycoside may be in a form in which the fructose is linked to the steviol glycoside via a β - (2,6) bond.

In particular, the transfructosylated steviol glycoside may be in a form in which from 1 to 3 fructose molecules are linked to the steviol glycoside.

In particular, the transfructosylated steviol glycoside may be transfructosylated stevioside or transfructosylated rebaudioside a. Thus, sweetener compositions of the present disclosure may include transfructosylated stevioside or transfructosylated rebaudioside a, or transfructosylated stevioside and transfructosylated rebaudioside a.

A sweetener composition comprising transfructosylated stevioside and transfructosylated rebaudioside A may contain 40 parts by weight or more of transfructosylated rebaudioside A, based on 100 parts by weight of the total weight of transfructosylated stevioside and transfructosylated rebaudioside A. Specifically, the composition may contain 50 parts by weight or more, more specifically 60 parts by weight or more, even more specifically 70 parts by weight or more, even more specifically 75 parts by weight or more of transfructosylated rebaudioside a.

In particular, the transfructosylated steviol glycosides may have an improved bitterness compared to steviol glycosides. An improvement in bitterness may mean a reduction in bitterness.

In particular, when transfructosylated steviol glycosides and steviol glycosides have the same sweetness, the improvement of bitterness can be evaluated. In particular, the sweetness may be that of sugar corresponding to 10 brix.

In particular, the transfructosylated steviol glycosides may have an improved sweetness compared to steviol glycosides. The improvement in sweetness may be an increase in sweetness preference and overall preference.

In particular, when transfructosylated steviol glycosides and steviol glycosides have the same sweetness, the improvement in sweetness can be evaluated. In particular, the sweetness may be that of sugar corresponding to 10 brix.

Specifically, transfructosylated steviol glycosides can be produced by reacting steviol glycosides with a fructose donor in the presence of a fructosyltransferase, but for the purposes of this disclosure, the reaction is not limited as long as transfructosylated steviol glycosides are obtained.

Specific nucleotide sequences of genes encoding fructosyltransferases and protein information thereof can be obtained from known databases such as GenBank of NCBI and the like. However, any protein may be included in addition to the above-mentioned known sequence as long as it exhibits the same effect of transferring fructose as a fructosyltransferase, without being limited by its origin or sequence. Furthermore, homologous or variant proteins may also be included within the scope of the fructosyltransferases of the present disclosure.

The step of reacting steviol glycoside with fructose donor in the presence of an enzyme may be performed at a pH of 3 to 8, specifically 4 to 7, more specifically 5 to 6, but the pH range is not limited thereto.

Further, the above step may be performed at 10 ℃ to 60 ℃, specifically at 20 ℃ to 50 ℃, more specifically at 20 ℃ to 40 ℃, but the temperature range is not limited thereto.

The sweetener compositions of the present disclosure can be used to prepare sweeteners or to provide sweetness to edible products. The sweetener compositions of the present disclosure may be used as sweeteners for the purpose of cooking and/or processing food products. Further, the sweetener compositions of the present disclosure may be used as sweeteners for pharmaceutical products in addition to edible products, but are not limited thereto. In addition, the sweetener compositions of the present disclosure may also include flavoring agents, preservatives, stabilizers, antioxidants, and the like, but are not limited thereto.

To achieve the object of the present disclosure, another aspect of the present disclosure provides a food composition comprising the sweetener composition.

The food compositions of the present disclosure may be prepared using methods common in the art, and may be prepared by adding raw materials and ingredients common in the art during the preparation process. In addition, the food composition of the present disclosure uses food as a raw material, unlike a counterfeit medicine, and thus has no side effects that may occur during long-term administration and has high portability.

The food composition may comprise the sweetener composition in an amount of 0.001 to 25 wt%, specifically 0.01 to 20 wt%, more specifically 0.01 to 10 wt%, based on the total weight of the composition, but is not limited thereto.

To achieve the object of the present invention, another aspect of the present invention provides a method for improving sweetness of steviol glycoside, which comprises converting steviol glycoside into transfructosylated steviol glycoside.

"steviol glycosides" and "transfructosylated steviol glycosides" are as described above.

For the purposes of this disclosure, the conversion is not limited by its method, as long as the transfructosylated steviol glycoside can be obtained from steviol glycoside.

Specifically, the conversion may include, but is not limited to, reacting steviol glycosides with a fructose donor in the presence of a fructosyltransferase.

The fructose donor may be any oligomeric, polymeric or cyclic form of fructose, which may be reacted in the presence of a fructosyltransferase such that one or more fructose molecules may be transferred to the steviol glycoside, in particular, it may be a sugar, but is not limited thereto.

Specific nucleotide sequences of genes encoding fructosyltransferases and protein information thereof can be obtained from known databases such as GenBank of NCBI and the like. However, any sequence may be included in addition to the above known sequences as long as it exhibits the same effect of transferring fructose as a fructosyl transferase, and is not limited to its source or sequence, and homologous or variant proteins may also be included within the scope of the fructosyl transferase of the present disclosure.

The step of reacting steviol glycoside with fructose donor can be carried out at a pH of 3 to 8, specifically at a pH of 4 to 7, more specifically at a pH of 5 to 6, but the pH range is not limited thereto.

In addition, the above step may be performed at 10 ℃ to 60 ℃, specifically at 20 ℃ to 50 ℃, more specifically at 20 ℃ to 40 ℃, but the temperature range is not limited thereto.

As used herein, the term "sweetness improvement" may refer to an improvement in sweetness preference or overall preference as bitterness or offensive odor/off-taste decreases.

Hereinafter, the present disclosure will be described in detail in connection with the attached exemplary embodiments. However, the exemplary embodiments disclosed herein are not for illustrative purposes only and should not be construed to limit the scope of the present disclosure.

Preparation example 1: preparation of transfructosylated steviol glycosides and transfructosylated rebaudiosides A

5% stevioside (Carbosynth) or 5% rebaudioside A (Purecyclie) and 20% white sugar (CJ CheilJedang) were dissolved in 50mM sodium acetate buffer, and then fructosyltransferase derived from Arthrobacter globiformis was added thereto, and the mixture was reacted at 40 ℃ for 24 hours. Subsequently, the mixture was adsorbed onto an adsorption resin (HP-20) and desorbed with 20% ethanol. Thereafter, the desorbed solution was concentrated under reduced pressure and dried to prepare a sample.

Fructosyltransferases transfer fructose from sugars. In particular, the resulting transfructosylated product is in the form of a glucose in which 1 to 3 fructose molecules are linked (transferred) by β - (2,6) bonds to the 19-OH position of stevioside or rebaudioside a or conjugated thereto.

Subjecting the structures of transfructosylated stevioside and transfructosylated rebaudioside A thus prepared to¹H/¹³C NMR, homonuclear correlation spectrum (COSY), Total correlation spectrum (TOCSY), Heteronuclear Single Quantum Coherence (HSQC), and Heteronuclear Multiple Bond Correlation (HMBC) were analyzed, and the results (1H/13C NMR, COZY, and HMBC) are shown in tables 1 and 2.

Furthermore, after confirming the structures of transfructosylated stevioside and transfructosylated rebaudioside a, the results show: transfructosylated stevioside was found to be 13- [ (2-O- β -D-glucopyranosyl- α -D-glucopyranosyl) oxy ] kauri-16-en-18-oic acid 6-O- β -D-fructofuranosyl- β -D-glucopyranosyl ester, and transfructosylated rebaudioside A was found to be 6-O- β -D-fructofuranosyl- β -D-glucopyranosyl-19-oxo-kauri-16-en-19-oic acid 6- [ (2-O- β -D-glucopyranosyl-3-O- β -D-glucopyranosyl) oxy ].

Transfructosylated stevioside and transfructosylated rebaudioside A are represented by chemical formula 4 and chemical formula 5, respectively.

[ chemical formula 4]

[ chemical formula 5]

Example 1: sensory descriptive analysis

The transfructosylated stevioside (STV-Fru) and transfructosylated rebaudioside a (RA-Fru) prepared in preparation example 1 were diluted to the sweetness equivalent to Stevioside (STV) and Rebaudioside A (RA), respectively (sweetness based on 10 brix sugars), and then presented to an expert (n-15) for descriptive analysis.

The results of comparison of transfructosylated rebaudioside A (RA-Fru) and Rebaudioside A (RA) are shown in Table 3 and FIG. 1.

The results of comparison of transfructosylated stevioside (STV-Fru) and Stevioside (STV) are shown in Table 4 and FIG. 2.

[ Table 3]

	RA	RA-Fru	p-value
				Intensity of sweetness	3.2	3.1	0.70
Sweetness linger	2.9	2.6	0.41
				Intensity of bitterness	3.4	1.8	0.00
Sense of weight	2.9	2.7	0.49
				Malodour/odour intensity	3.3	2.4	0.01
Sweetness preference	2.5	3.4	0.00
				General preference	2.6	3.5	0.01

[ Table 4]

As shown in tables 3 and 4, it can be found that transfructosylated rebaudioside a and transfructosylated stevioside (STV-Fru) has significantly lower bitterness intensity and offensive odor/off taste intensity as well as significantly higher sweetness preference and overall preference as compared to rebaudioside a (ra) and Stevioside (STV).

Example 2: sensory preference evaluation

Four samples of example 1 (STV, STV-Fru, RA-Fru) were diluted to a sweetness equivalent to 10% (w/w) or 10 Brix sugar before sensory evaluation, and the four samples were subjected to sensory evaluation.

The evaluation was carried out as follows: four samples were given to 15 groups and the four samples were asked to be evaluated for sweetness preference and overall preference on a 9-level scale (1 ═ very dislike, 5 ═ neither dislike nor dislike, 9 ═ very like).

Statistical analysis was then converted to a 5-grade scale and significant differences were verified by paired T-test (confidence level 95%).

(1) Comparison between STV and STV-Fru

[ Table 5]

Sensory attributes	STV	STV-Fru	P-value
				Sweetness preference	2.13	3.14	0.0002
General preference	2.02	3.02	0.001

Table 5 shows that transfructosylated stevioside (STV-Fru) has superior sweetness preference and overall preference compared to Stevioside (STV).

(2) Comparison between RA and RA-Fru

[ Table 6]

Table 6 shows that transfructosylated rebaudioside a (RA-Fru) has superior sweetness preference and overall preference compared to Rebaudioside A (RA).

PREPARATION EXAMPLE 2 preparation of a mixture comprising Transfructosylated stevioside and Transfructosylated rebaudioside A

A mixture of 5% stevioside (Carbosynth) and 5% rebaudioside A (Pureccle) in which the proportions of rebaudioside A in the mixture were 4 wt%, 60 wt%, 80 wt%, respectively, and 20% white sugar (CJ CheilJedang) was dissolved in 50mM sodium acetate buffer, and then a mixture comprising transfructosylated stevioside and transfructosylated rebaudioside A was prepared in the same manner as in preparation example 1. The ratio of transfructosylated stevioside to transfructosylated rebaudioside A in the mixture so prepared was maintained at the same level as the ratio of stevioside to rebaudioside A in the starting material.

Although the present disclosure has been described with reference to specific illustrative embodiments, those skilled in the art to which the present disclosure pertains will appreciate that the present disclosure may be embodied in other specific forms without departing from the technical spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive. Further, the scope of the present disclosure is defined by the appended claims rather than the detailed description, and it should be understood that all modifications or variations derived from the meaning and scope of the present disclosure and equivalents thereof are included in the scope of the appended claims.