Title of Invention: Composition for Inhibiting HMF Generation Containing Allulose Disaccharide
technical field
[One]
The present application relates to novel uses of allulose disaccharides.
[2]
background
[3]
HMF is an organic compound formed by dehydration of a compound containing an aldehyde group and a hydroxyl group, and is a white low-melting compound that is highly soluble in both water and organic solvents. The large amount of HMF production suggests that there is a high possibility of inducing continuous non-enzymatic browning, as well as quality deterioration due to deterioration of freshness or accumulation of heat damage during product processing and distribution. Moreover, there are still unidentified negative concerns from the point of view of human risk, such as the issue of carcinogenicity of HMF (Abraham K (2011) Toxicology and risk assessment of 5-Hydroxymethylfurfural in food, Molecular Nutrition & Food Research. 55 ( 5): 667-678).
[4]
Heat sterilization is the most commonly used method in the processing of food, but since HMF is generated when heated as described above in foods containing sugar, it is an important task to reduce the amount of HMF generated.
[5]
To solve this problem, research on product quality stabilization through suppression of HMF production is being conducted, and Food Browning and Its Prevention: An Overview. J. Agric. Food Chem., Vol. 44, No. Although it has been studied in 3 et al., there is a need for research on a method having a more excellent effect, and furthermore, a method of adding carbohydrates has not been reported yet.
[6]
DETAILED DESCRIPTION OF THE INVENTION
technical challenge
[7]
Under this background, the present inventors isolated a novel material, confirmed that this material is an allulose disaccharide, and confirmed that the production of HMF is inhibited when allulose disaccharide is added to a composition containing saccharides, thereby completing the present invention. .
[8]
means of solving the problem
[9]
The present application provides a composition for inhibiting HMF (Hydroxymethylfurfural) production and/or preventing browning, including allulose disaccharide.
[10]
The present application provides a method for inhibiting HMF production and/or preventing browning, comprising preparing a composition comprising saccharides and allulose disaccharides.
[11]
The present application relates to preparing a mixed composition comprising saccharides and allulose disaccharides; And to provide a method for producing a composition comprising a saccharide, comprising heating the mixture composition.
[12]
Effects of the Invention
[13]
Since the composition containing allulose disaccharide of the present application suppresses the generation of harmful substances generated during processing, sterilization and long-term storage, it can be usefully used to suppress sugar dehydration reaction, suppress HMF production, and/or prevent browning.
[14]
Brief description of the drawing
[15]
1 is an HPLC chromatogram analyzed by column- (Biorad Aminex HPX-87C) of disaccharides produced during the preparation of allulose.
[16]
FIG. 2 is an HPLC chromatogram of D1 and D2 analyzed by column (YMC Pack Polyamine II) of a mixture of disaccharides produced during the production of allulose by column (YMC Pack Polyamine II).
[17]
3 shows the three-dimensional structure of D1, which is an allulose disaccharide.
[18]
4 shows the structure and carbon numbering of allulose.
[19]
Best mode for carrying out the invention
[20]
A detailed description of this is as follows. Meanwhile, each description and embodiment disclosed in the present application may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. In addition, it cannot be seen that the scope of the present application is limited by the specific descriptions described below.
[21]
In addition, those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the present application described herein. Also, such equivalents are intended to be covered by this application.
[22]
One aspect of the present application provides a composition for inhibiting HMF (Hydroxymethylfurfural) production comprising allulose disaccharide.
[23]
[24]
In the present application, "allulose disaccharide" may be used interchangeably with terms such as "allulose dimer", "allulose diploid", "disaccharide allulose", and "allulose bimolecule is glyco compounds linked by seed bonds".
[25]
Specifically, in the allulose disaccharide, two molecules of allulose are linked by a glycosidic bond, and the glycosidic bond is a hydroxyl group of carbon 2 (C2) of one molecule of allulose among the two molecules of allulose. It may be a glycosidic bond bonded to a hydroxyl group of any one of carbons 1 to 6 (C1 to C6) of one molecule of other allulose.
[26]
Specifically, at least one molecule of two molecules of allulose is cyclic allulose, and a hydroxyl group at carbon 2 of the cyclic allulose and a hydroxyl group at any one carbon of carbon 1 to 6 of another allulose molecule It may be a compound connected between them by a glycosidic bond. The number of glycosidic bonds may be 1 to 2, specifically, 1 may be.
[27]
In one embodiment, the bond may be a glycosidic bond between a hydroxyl group on carbon 2 of cyclic allulose and a hydroxyl group on carbon 6 of another allulose.
[28]
In one embodiment, one molecule of the two molecules of allulose may be in the form of psicofuranose, and the other molecule may be in the form of psicopyranose. In one embodiment, it may be one represented by the following formula (1), but is not limited thereto.
[29]
[Formula 1]
[30]

[31]
In one embodiment, the allulose disaccharide of the present application is 2-(hydroxymethyl)-2-((3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methyl oxy)tetrahydro-2H-pyran-3,4,5-triol(2-(hydroxymethyl)-2-((3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro -2H-pyran-3,4,5-triol), and more specifically (2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S) ,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol ((2S ,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H -pyran-3,4,5-triol) may be a compound named, but is not limited thereto.
[32]
In one embodiment, the allulose disaccharide of the present application is 2-(hydroxymethyl)-2-((3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methyl oxy)tetrahydro-2H-pyran-3,4,5-triol(2-(hydroxymethyl)-2-((3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro -2H-pyran-3,4,5-triol), and more specifically (2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S) ,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol ((2S , 3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H -pyran-3,4,5-triol) may be a compound named, but is not limited thereto.
[33]
(2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydro Furan-2-yl) methoxy) tetrahydro-2H-pyran-3,4,5-triol is 6-O-β-D-psychopyranosyl-α-D-psycho, depending on the form of psychofuranose Furanose (6-O-β-D-Psicopyranosyl-α-D-psico furanose) or 6-O-β-D-Psycopyranosyl-β-D-Psicopyranosyl -β-D-psico furanose) may be collectively named.
[34]
(2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydro Furan-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol is (2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R) ,3S,4R,5S)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-tri All((2S, 3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R,5S)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl )methoxy)tetrahydro-2H-pyran-3,4,5-triol), or (2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R) ,3S,4R,5R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-tri All((2S, 3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R,5R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl )methoxy)tetrahydro-2H-pyran-3,4,5-triol), but is not limited thereto.
[35]
Specifically, the compound of Formula 1 may exist in two forms of Formulas 2 and/or 3 below.
[36]
[Formula 2]
[37]

[38]
[Formula 3]
[39]

[40]
The compound of Formula 2 is 6-O-β-D-Psycopyranosyl-α-D-Psycofuranose (6-O-β-D-Psicopyranosyl-α-D-psico furanose), and the compound of Formula 3 is 6-O-β-D-Psicopyranosyl-β-D-Psycofuranose (6-O-β-D-Psicopyranosyl-β-D-psico furanose).
[41]
[42]
"HMF" in the present application is a material also referred to as 5'-HMF (5-hydroxymethyl-furfural), and may have a structure represented by Chemical Formula 4 below.
[43]
[Formula 4]
[44]

[45]
The HMF may be generated from a compound containing an aldehyde group and a hydroxyl group.
[46]
In the present application, "HMF-generating material" means a compound capable of generating HMF.
[47]
The HMF-generating material includes without limitation as long as it includes an aldehyde group and/or a hydroxyl group, and for example, may also include materials such as carbohydrates, glycolipids, and glycoproteins.
[48]
Specifically, the HMF generation may be due to saccharides (expressed as sugar or saccharide). The HMF production may be due to glycolysis. The decomposition of sugar includes decomposition of sugar by dehydration.
[49]
Specifically, the saccharide may be a monosaccharide. The monosaccharides include, aldotriose (glyceraldehyde), ketotriose (dihydroxyacetone), aldotetrose (erythrose, threose), ketotetrose (erythrulose), aldopentose (arabinose, Rixose, ribose, xylose (wood sugar, wood sugar)), ketopentose (ribulose, xylulose), deoxysaccharide (deoxyribose), aldohexose (allose, altrose, galactose, glucose (glucose) ), gulose, idos, mannose, talose), ketohexose (fructose (fructose), allulose, sorbose, tagatose), deoxysaccharides (fucose, fuculose, rhamnose), ketoheptose (mannoheptulose, sedoheptulose), etc., if there is a possibility of generating HMF, includes without limitation. Specifically, it may be a hexose sugar, and more specifically, it may be allulose. However, it is not limited thereto.
[50]
Meanwhile, a mixture including one or more of the above-mentioned substances may also be included in the HMF-generating material.
[51]
The HMF is also an example of a glycation intermediate. As used herein, the term “glycation product” refers to a product produced by a reaction between an amino acid group such as a lysine residue of a protein and a reducing sugar without the action of an enzyme, and includes both a glycation intermediate product and a glycation final product, and includes glycation intermediate products (glycation intermediate products) intermediates) as glycation endproducts (AGEs). The final product of saccharification may be brown in color and generate volatile aromatic components, or may refer to various substances produced by reacting with various protein components such as hemoglobin, LDL, collagen, etc. with blood glucose or degradation products of glucose in the body. have. The saccharification product corresponds to a representative example of a by-product generated in the processing, storage, and sterilization reaction of a composition containing saccharides.
[52]
Once the final glycated product is formed, it is not decomposed even when blood sugar is restored to normal and accumulates in the blood or tissue during the protein's survival period. Accumulated end-glycosylation products form cross-links with proteins and interact with receptor for AGEs (RAGEs), leading to accumulation of inflammatory cells.
[53]
Therefore, the production of the final product of saccharification and the amount of HMF, which can have a detrimental effect on the human body, are closely related. The allulose disaccharide of the present application may also inhibit the production of the final product of saccharification by inhibiting the production of HMF. .
[54]
That is, another aspect of the present application provides a composition for inhibiting the production of saccharification products including allulose disaccharide.
[55]
[56]
Another aspect of the present application provides a composition for inhibiting sugar dehydration reaction (hydration) comprising allulose disaccharide.
[57]
[58]
As used herein, the term “hydration” refers to the entire process in which water is separated within or between molecules. In the present application, the molecule causing the dehydration reaction may be a sugar molecule.
[59]
In the present application, "sugar dehydration reaction" refers to a reaction in which H 2 O is generated within a sugar molecule or between sugar molecules . The sugar dehydration reaction, specifically, may be a reaction in which H 2 O is generated in a sugar molecule .
[60]
Specifically, the sugar molecule may be a monosaccharide, which is a unit of sugar (glucide). The monosaccharide is the same as described above.
[61]
When a dehydration reaction occurs in sugar molecules, other substances derived from sugar molecules may be produced in addition to H 2 O molecules. As an example, the dehydration reaction in the hexose may be a reaction for generating hydroxymethylfurfural, that is, HMF, in addition to H 2 O molecules as shown in Scheme 1 below.
[62]
[63]
[Scheme 1]
[64]

[65]
Suppression of the sugar dehydration reaction means preventing the above-mentioned sugar dehydration reaction from occurring or reducing the sugar dehydration reaction from occurring compared to an environment in which allulose disaccharide is not present or a relatively small amount is present. Inhibition of the sugar dehydration reaction can be confirmed by measuring the amount of the sugar dehydration reaction product. For example, by measuring the amount of HMF production, it can be confirmed whether the sugar dehydration reaction is inhibited.
[66]
[67]
The dehydration reaction may occur under heating, sterilization and/or processing conditions of a known composition, but is not limited thereto, and also includes a reaction that occurs naturally at room temperature, so a reaction that occurs during storage of the composition is also included.
[68]
[69]
Another aspect of the present application provides a composition for preventing browning comprising allulose disaccharide.
[70]
In the present application, "anti-browning" may be used to mean preventing browning from occurring, delaying browning, inhibiting browning, and the like, and may be used interchangeably in the present application. For example, the browning reaction in foods such as cereals and cereal bars, potato chips, bakery, carbonated beverages, fruit and vegetable juices, fruit juices, fruit wines, sauces, candy, jellies, jams, ice cream, beer, etc. has aroma, taste and nutritional value. deterioration of quality leading to the loss of
[71]
A Maillard reaction, a caramelization reaction, etc. may appear as a browning reaction. For example, in the Maillard reaction, a carbonyl group of a saccharide and an amino acid group of a protein may react by heating or the like to generate brown substances (melanoidins). This reaction is also called the melanoidin reaction after the reactant and is also called the aminocarbonyl reaction due to the reactant.
[72]
In the intermediate stage of the Maillard reaction, HMF, a highly reactive substance, is generated, and the products in the final stage of the reaction are also highly reactive substances. These substances form a polymer to form a brown fluorescent melanoidin pigment. In this process, browning takes place.
[73]
As described above, since the generation of HMF is closely related to the browning reaction of the composition, the allulose disaccharide of the present application may be used for anti-browning purposes by inhibiting the generation of HMF. That is, the browning prevention may be due to inhibition of HMF production.
[74]
[75]
The content of allulose disaccharide in the composition may include allulose disaccharide in an amount of more than 0 and 15 parts by weight or less based on 100 parts by weight of the HMF-producing material included in the composition.
[76]
Specifically, it contains more than 0.0001 parts by weight, more than 0.001 parts by weight, more than 0.01 parts by weight, more than 0.1 parts by weight, or more than 0.15 parts by weight and not more than 15 parts by weight of allulose disaccharide based on 100 parts by weight of the total HMF-producing material. and/or 15 parts by weight or less, 13 parts by weight or less, 11 parts by weight or less, 10 parts by weight or less, 9 parts by weight or less, 8 parts by weight or less, 7 parts by weight or less, of allulose disaccharide relative to 100 parts by weight of the HMF-producing material It may be included in an amount of parts by weight or less, 6 parts by weight or less, 5 parts by weight or less, 4 parts by weight or less, 3 parts by weight or less, 2 parts by weight or less, or 1 part by weight and more than 0 parts by weight, but is not limited thereto.
[77]
The content of allulose disaccharide in the composition may include allulose disaccharide in an amount of more than 0 and 15 parts by weight or less based on 100 parts by weight of the total saccharides included in the composition. Specifically, it may contain allulose disaccharides in an amount of more than 0.0001 parts by weight, more than 0.001 parts by weight, more than 0.01 parts by weight, more than 0.1 parts by weight, or more than 0.15 parts by weight and not more than 15 parts by weight, based on 100 parts by weight of the total saccharides, , and/or, 15 parts by weight or less, 13 parts by weight or less, 11 parts by weight or less, 10 parts by weight or less, 9 parts by weight or less, 8 parts by weight or less, 7 parts by weight or less, allulose disaccharide relative to 100 parts by weight of total saccharides; 6 parts by weight or less, 5 parts by weight or less, 4 parts by weight or less, 3 parts by weight or less, or 2 parts by weight or less and may be included in an amount exceeding 0 parts by weight, but is not limited thereto.
[78]
Alternatively, the content of allulose disaccharide in the composition may include allulose disaccharide in an amount of more than 0 and 15 parts by weight or less based on 100 parts by weight of the total solids contained in the composition. Specifically, more than 0.0001 parts by weight, more than 0.001 parts by weight, more than 0.01 parts by weight, more than 0.1 parts by weight, or more than 0.15 parts by weight and not more than 15 parts by weight of allulose disaccharide relative to 100 parts by weight of the total solid content. Or, 15 parts by weight or less, 13 parts by weight or less, 11 parts by weight or less, 10 parts by weight or less, 9 parts by weight or less, 8 parts by weight or less, 7 parts by weight or less, 6 parts by weight or less, allulose disaccharide relative to 100 parts by weight of total solid content Parts by weight or less, 5 parts by weight or less, 4 parts by weight or less, 3 parts by weight or less, or 2 parts by weight or less, but may be included in an amount exceeding 0 parts by weight, but is not limited thereto.
[79]
[80]
The composition may be a food composition. The food composition of the present application includes, but is not limited to, general food, health food, and medical (or patient) food composition. Specifically, the food composition of the present application is a beverage (eg, carbonated beverage, fruit juice, fruit and vegetable beverage, dietary fiber beverage, carbonated water, wheat flour, tea, coffee, etc.), alcoholic beverage, bakery, sauce (eg, ketchup, pork cutlet sauce) etc.), dairy products (eg, fermented milk, processed milk, etc.), processed meat products (eg, ham, sausage, jerky, etc.), chocolate products, gum, candy, jelly, ice cream, syrup, dressing, snacks (eg, cookies, crackers, biscuits, etc.) etc.), pickled fruits and vegetables (eg, cheong, dangchim fruit, red ginseng extract or red ginseng slices, etc.), meal substitutes (eg, frozen food, retort food, home meal replacement (HMR), etc.) or processed food. However, this is only an example and is not limited thereto.
[81]
The food composition of the present application may contain various flavoring agents or natural carbohydrates as additional ingredients. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, synthetic sweeteners such as sucralose, saccharin, and aspartame may be used.
[82]
In addition to the above, the food composition of the present application includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectin and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, It may contain acidifying agents and salting agents used in carbonated beverages. In addition, the food composition of the present application may contain flesh for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. In addition, a person skilled in the art may appropriately select and add substances that may be normally included in the food composition, and the ratio of these additives may be selected in the range of 0.001 to 1 parts by weight, or 0.01 to 0.20 parts by weight per 100 parts by weight of the food composition of the present application. can, but is not limited thereto.
[83]
[84]
Another aspect of the present application provides a method for inhibiting HMF production, comprising preparing a composition comprising saccharides and allulose disaccharides.
[85]
"Prepare" in the present application includes, without limitation, any method of providing a composition comprising saccharides and allulose disaccharides. That is, it includes any method that allows the composition to contain saccharides and allulose disaccharides. For example, preparing a composition comprising saccharide and allulose disaccharide includes adding allulose disaccharide to a composition including saccharide, adding saccharide to a composition including allulose disaccharide, and preparing saccharide/saccharide composition sia allulose disaccharides are produced, and the like.
[86]
As long as the composition includes saccharides and allulose disaccharides, other components may be included without limitation.
[87]
Meanwhile, a composition comprising saccharides and allulose disaccharides may also be referred to as a “mixture”. In the mixed composition, "saccharides" other than allulose disaccharides may include, for example, allulose, but is not limited thereto.
[88]
The method for inhibiting HMF production may further include heating the composition after preparing the composition including saccharides and allulose disaccharides. However, the heating is not limited thereto, and the heating may be performed before, after, or simultaneously with the preparation of the mixed composition.
[89]
The heating may be performed in an appropriate temperature range depending on the type of composition, and the temperature range, heating time, sterilization method, etc. may be appropriately performed by those skilled in the art based on information known in the art. Specifically, it may be carried out at a temperature of 60 ℃ or more and 100 ℃ or less, and more specifically, it may be carried out at a temperature of 60 ℃ or more and 95 ℃ or less, 65 ℃ or more and 95 ℃ or less, 70 ℃ or more and 95 ℃ or less. not limited
[90]
The heating may be performed for more than 0 hours and 108 hours or less, specifically, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours. , 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours or more may be performed, but is not limited thereto.
[91]
Another aspect of the present application is to prepare a mixed composition comprising saccharides and allulose disaccharides; and heating the mixed composition.
[92]
Heating the mixture composition may inhibit HMF production, but is not limited thereto.
[93]
The heating may be performed before, after, or simultaneously with the preparation of the mixed composition.
[94]
The composition containing the saccharide prepared by the above preparation method has a low impurity content, a small amount of HMF, an increase in allulose content, a change in its physical properties, a by-product, crystallization, browning reaction, oxidation / Reduction reaction, a reaction in which saccharides other than allulose disaccharide are converted into other substances, etc. may occur less frequently. Specifically, compared to the case of heating a composition that does not contain allulose disaccharide or has a relatively low content of allulose disaccharide compared to the mixed composition under the same conditions, the above-described reaction may occur less. However, it is not limited thereto.
[95]
[96]
Another aspect of the present application provides a method for inhibiting sugar dehydration, comprising preparing a composition comprising saccharides and allulose disaccharides.
[97]
Another aspect of the present application provides a method for preventing browning, comprising preparing a composition comprising saccharides and allulose disaccharides.
[98]
Another aspect of the present application provides a method for sterilizing a composition, comprising preparing a composition comprising a saccharide and an allulose disaccharide.
[99]
The method may further comprise heating the composition after preparing the composition comprising the saccharide and allulose disaccharide. However, the heating is not limited thereto, and the heating may be performed before, after, or simultaneously with the preparation of the mixed composition.
[100]
[101]
HMF production, sugar dehydration, browning prevention, saccharides, and heating are the same as described above.
[102]
The composition may be, for example, a food composition, but is not limited thereto.
[103]
Food is the same as described above.
[104]
[105]
Another aspect of the present application provides a composition for inhibiting saccharide denaturation comprising allulose disaccharide.
[106]
Another aspect of the present application provides a method for inhibiting denaturation of a composition, comprising preparing a composition comprising saccharides and allulose disaccharides.
[107]
The method may further comprise heating the composition after preparing the composition comprising the saccharide and allulose disaccharide. However, without being limited thereto, the heating may be performed before, after, or simultaneously with the preparation of the mixed composition.
[108]
Allulose disaccharides, saccharides, and heating are as described above.
[109]
The denaturation includes conversion of saccharides to other substances, such as crystallization, browning reaction, and oxidation/reduction reaction, changes in their physical properties, or generation of by-products. However, it is not limited thereto.
[110]
[111]
Another aspect of the present application provides a use for inhibiting HMF production of allulose disaccharide.
[112]
The inhibition of allulose disaccharide and HMF production is the same as described above.
[113]
Modes for carrying out the invention
[114]
Hereinafter, the present application will be described in more detail through Examples and Experimental Examples. However, these Examples and Experimental Examples are for illustrative purposes of the present application, and the scope of the present application is not limited to these Examples and Experimental Examples.
[115]
[116]
Example 1: Isolation of novel allulose disaccharides
[117]
In the process of preparing allulose known from US 2018-0327796 A1, disaccharides were separated through HPLC. Specifically, the HPLC chromatogram analysis conditions shown in Table 1 below were performed, and it was confirmed that, as shown in FIG. 1 , a new unknown substance other than allulose was generated from the stock solution.
[118]
Although the content of the new material separated as described above was somewhat different depending on the manufacturing process, it was present at 2% or less in the initial stock solution, and it was confirmed that it increased to a level of 5% depending on the storage time.
[119]
[120]
[Table 1]
Equipment Agilent technologies 1200 series
Column Biorad Aminex HPX-87C (7.8 X 300mm, 9um)
Eluent Water
flow rate 0.6mL/min
Temperature 80℃
RI cell temperature 35℃
[121]
[122]
As a result, allulose was confirmed at 21.1 minutes and a novel substance was identified at 31.7 minutes.
[123]
Accordingly, in order to separate the generated new material, the novel material was purified to a purity of 95% or more using preparative HPLC, and again precisely separated using a normal phase column.
[124]
Specifically, HPLC chromatograms were performed.
[125]
Chromatogram separation conditions are shown in Table 2 below.
[126]
[127]
[Table 2]
Equipment Shimadzu LC 10A
Column YMC Pack Polyamine II (4.6 X 250mm, 5um,12nm)
Eluent Acetonitrile / Water (80/20)
flow rate 1mL/min
Temperature 30℃
RI cell temperature 30℃
[128]
[129]
As a result, it was confirmed that the material that appeared as one peak under the HPLC conditions of Table 1 appeared as two peaks under the separation conditions of Table 2 (FIG. 2), and the material of the peak identified at 22.5 minutes was confirmed at D1 and 17.7 minutes. The material at the peak was named D2.
[130]
[131]
Example 2: Confirmation of HMF production inhibitory effect of allulose disaccharide
[132]
Example 2-1: Comparison of HMF production rate of allulose disaccharide
[133]
Allulose was selected as a representative example of a monosaccharide that was denatured upon heating and produced high HMF. In order to confirm that the disaccharide (dimer) separated in Example 1 can be applied to various types of foods having different amounts of saccharide, the difference was compared by varying the saccharide concentration. In addition, considering the harsher environment, HMF production rates were compared in a retort environment (121°C, 15 minutes) with the highest temperature among food sterilization conditions.
[134]
Specifically, ultrapure water without impurities is added using crystalline allulose (CJ Cheiljedang, purity of 99% or more) with the highest composition ratio as a monosaccharide, and the disaccharides separated in Example 1 with a purity of 95% or more are quantitatively measured to determine the mixing ratio. Differently, Experimental Example 1 was prepared. In addition, samples (A) to (F) were prepared so that the concentrations of 1, 5, 10, 20, 30, and 50% (w/w) were respectively different by varying the amount of water added in Experimental Example 1 (Table 3) .
[135]
Meanwhile, in order to compare the effects of other disaccharides, sugar, a representative disaccharide, was added instead of allulose disaccharide and used as Comparative Example 1. Specifically, in the same manner as in Experimental Example 1, sugar, a disaccharide, was added to the monosaccharide allulose crystal in the same ratio as in Experimental Example 1, and the ratio of the components was confirmed using HPLC under the conditions of Table 1. Comparative Example 1 and Experimental Example 1, in which the ratio of the components was confirmed, was prepared by dissolving in ultrapure water without impurities to 1 to 50% (w/w).
[136]
[137]
[Table 3]
Sample name Constituent sugars
(based on solids weight of 100) mixing ratio Concentration (%, w/w)
Monomer
(Allulose) Dimer M:D (A) (B) (C) (D) (E) (F)
Comparative Example 1 95.214 1.904 50:1 One 5 10 20 30 50
Experimental Example 1 94.855 1.901 50:1 One 5 10 20 30 50
[138]
[139]
All prepared samples were placed in an autoclave (Jeotech, ST-105G) and heated at 121°C for 15 minutes. At this time, the time taken for the temperature increase of the equipment was not considered, and the heating time was measured after reaching the target temperature. After the heat treatment was completed, the sample was taken out and left at room temperature for 10 minutes, and then analyzed using HPLC under the conditions shown in Table 1 of Example 1.
[140]
All experiments were repeated three times, and the results are shown in Table 4 below.
[141]
[142]
[Table 4]
Sample name Concentration
(%, w/w) Constituent sugars (based on solids weight of 100) HMF increase rate
Monomer
(Allulose) Dimer %
Comparative Example 1 (A) One 93.310 1.855 182.8%
(B) 5 89.406 1.847 458.7%
(C) 10 94.167 1.897 291.5%
(D) 20 92.072 1.866 337.8%
(E) 30 92.167 1.897 378.7%
(F) 50 90.453 1.872 577.5%
Experimental Example 1 (A) One 97.516 0.765 114.1%
(B) 5 98.565 0.431 128.8%
(C) 10 98.560 0.449 144.7%
(D) 20 98.185 0.576 156.6%
(E) 30 97.524 0.797 219.9%
(F) 50 95.695 1.533 285.4%
[143]
As a result of the experiment, in Experimental Example 1 containing a certain amount of allulose disaccharide, it was confirmed that the increase rate of HMF was significantly small. Although the heating temperature was very high, it was not possible to completely block the generation of HMF, but compared to Comparative Example 1, in which sugar was added in the same ratio instead of allulose disaccharide as a disaccharide, the monosaccharide directly absorbed heat damage and rapidly decomposed. It is confirmed that the stability is improved. That is, it can be confirmed that compared with other disaccharides, allulose disaccharide has a significantly higher effect of delaying and protecting monosaccharides from deterioration by heating.
[144]
In addition, from the viewpoint of concentration, it was confirmed that the higher the concentration sample, the greater the absolute amount of monosaccharide, which is the HMF-generating material, the higher the generation of HMF under the same heating condition. However, compared to Comparative Example 1 in which sugar was added, it was confirmed that in Experimental Example 1 in which allulose disaccharide was added, the generation of HMF was significantly less and the denaturation of monosaccharides was delayed.
[145]
Through this, it was confirmed that allulose disaccharide inhibits dehydration, decomposition, and denaturation of monosaccharides even in a harsh environment at very high temperature, so that allulose disaccharide can be usefully used to suppress the production of HMF and suppress the dehydration of saccharides.
[146]
[147]
Example 2-2: Comparison of HMF production rate by allulose disaccharide content
[148]
Disaccharides (dimers) separated in Example 1 were mixed as shown in Table 5 below to prepare samples having different ratios of disaccharides.
[149]
Specifically, as a monosaccharide, ultrapure water without impurities is added using crystalline allulose with the highest composition ratio (CJ Cheiljedang, purity of 99% or more) to 10% (w/w) similar to the average concentration of conventional beverages. prepared and used as Experimental Example 2. In addition, allulose disaccharides with a purity of 95% or more isolated in Example 1 were quantitatively measured and added to crystalline allulose, and those prepared by dissolving in ultrapure water to a concentration of 10% were used in Experimental Examples 3 to 4. The composition of each prepared sample was analyzed once again under the conditions of Table 1 of Example 1 using HPLC, and it was confirmed that the amount of disaccharide included was different as shown in Table 5 below.
[150]
[151]
[Table 5]
Sample name Constituent sugars
(based on solids weight of 100) mixing ratio density
Monomer Dimer Others Monomer: Dimer (%, w/w)
Experimental Example 2 99.823 0.158 0.019 632:1 10
Experimental Example 3 97.532 1.154 1.314 85:1 10
Experimental Example 4 95.41 2.145 2.614 44:1 10
[152]
[153]
Each prepared sample was heated at 95° C., which is a typical process temperature for beverages, and sampled at 20-minute intervals to check the change in components and the amount of HMF produced. Quantification of HMF was analyzed using HPLC under the conditions of Table 1 of Example 1.
[154]
[155]
All experiments were repeated three times, and the results are shown in Table 6 below.
[156]
[157]
[Table 6]
Sample name Heating time
(min, 95℃) Constituent sugars (based on solids weight of 100) HMF increase rate
Monomer Dimer %
Experimental Example 2 0 99.8a 0.2a 100.0%d
20 99.8b 0.2b 110.3%c
40 99.7c 0.1c 120.5%b
60 99.7d 0.1d 128.2%a
p 0.000 0.000 0.000
Experimental Example 3 0 97.5d 1.2a 100.0%d
20 98.1c 0.9b 103.5%c
40 98.5b 0.6c 108.8%b
60 99.1a 0.3d 112.3%a
p 0.000 0.000 0.000
Experimental Example 4 0 95.2d 2.1a 100.0%a
20 96.3c 1.6b 100.0%a
40 97.4b 1.0c 101.3%a
60 98.5a 0.4d 101.3%a
p 0.000 0.000 0.110
[158]
※ Strings a,b,c,d different in the vertical direction mean that there is a significant difference (p<0.05) according to the heating time in the same sample.
[159]
[160]
In Experimental Examples 2 to 4 in which a certain amount of disaccharide was included, it was confirmed that a sufficient amount of disaccharide first absorbed heat damage and, in the process of decomposition, rather increased monosaccharide. Accordingly, it was confirmed that the increase rate of HMF generated from monosaccharides was also significantly small. In particular, in the case of Experimental Example 4 containing the most disaccharides (including 2.1% (w/w) of the constituent sugars and about 0.21% (w/w) based on the total amount of the sample), the HMF increase rate was statistically significant after 60 minutes of heating. It was confirmed that there was no difference, and it was confirmed that the effect of inhibiting the generation of HMF was very large.
[161]
Through this, it was confirmed that allulose disaccharide inhibits dehydration, decomposition, and denaturation of monosaccharides and suppresses the generation of HMF. It can be seen that the quality deterioration phenomenon can be extremely delayed.
[162]
[163]
Through this experimental process, it was confirmed that when a certain amount of disaccharide including allulose was contained, the saccharide (allulose) was decomposed and denatured by heat to significantly delay the generation of HMF.
[164]
Compared to the conventional use of additives composed of completely different ingredients (eg, additives such as antioxidants) to suppress HMF production, disaccharides based on saccharides are used, so the effect on the taste and characteristics of the product is extremely It has the advantage that it can be reduced.
[165]
[166]
Example 3: Identification of the structure of allulose disaccharide
[167]
In order to confirm the structure of the allulose disaccharide having the function of inhibiting HMF production, the structures of D1 and D2 isolated in Example 1 were identified through ESI-MS, 1H NMR, and 13C NMR.
[168]
Specifically, the structure was identified through the following method.
[169]
[170]
Major 6-O-β-D-Psicopyranosyl-α-D-psicofuranose silver, white amorphous powder, ESI-MS m/z 365 [M+Na]+; 1H NMR (850 MHz, D2O) δH 3.44 (1H, d, J = 12.0 Hz), 3.47 (1H, d, J = 12.0 Hz), 3.56 (1H, dd, J = 11.0, 5.0 Hz), 3.60 (1H) , d, J = 12.0 Hz), 3.62 (1H, dd, J = 11.0, 2.5 Hz), 3.70 (1H, br d, J = 12.5 Hz), 3.75 (1H, d, J = 12.0 Hz), 3.75 ( 1H, br ma), 3.82 (1H, br d, J = 12.5 Hz), 3.84 (1H, br s), 3.92 (1H, t, J = 3.0 Hz), 3.97 (1H, d, J = 5.5 Hz) , 4.09 (1H, t, J = 5.5 Hz), 4.13 (1H, br m) [D2O signal δH 4.70]; 13C NMR signals δC 57.6, 60.4, 62.9, 64.7, 64.9, 69.1, 68.9, 70.2, 70.3, 81.2, 101.8, 103.4.
[171]
Minor 6-O-β-D-Psicopyranosyl-β-D-psicofuranose, white amorphous powder, ESI-MS m/z 365 [M+Na]+; 1H NMR (850 MHz, D2O) δH 3.49 (1H, d, J = 13.0 Hz), 3.73 (1H, d, J = 13.0 Hz), 3.58 (1H, ma), 3.68 (1H, dd, J = 11.0, 2.5 Hz), 3.62 (1H, ma), 3.71 (1H, br d, J = 12.0 Hz), 3.82 (1H, br d, J = 12.0 Hz), 3.76 (1H, br ma), 3.78 (1H, ma) ), 3.87 (1H, br s), 3.98 (1H, t, J = 3.0 Hz), 3.95 (1H, d, J = 4.5 Hz), 4.00 (1H, br m), 4.34 (1H, dd, J = 8.0, 4.5 Hz) [DO signal δH 4.70]; 13C NMR signals δC 57.7, 61.4, 62.2, 64.7, 64.8, 69.0, 69.2, 70.8, 74.4, 80.8, 101.8, 105.9.
[172]
[173]
[174]
As a result, it was confirmed that D1 is a novel allulose disaccharide and has the structure of Formula 1 below.
[175]
[Formula 1]
[176]

[177]
[178]
In addition, D1 has two forms of major or minor form (FIG. 3), and 6-O-β-D-Psicopyranosyl-α-D-psicofuranose, which is the major form, has the following formula 2, 6-O-β which is the minor form. -D-Psicopyranosyl-β-D-psicofuranose was confirmed to have the structure of Formula 3 below.
[179]
[180]
[Formula 2]
[181]

[182]
[183]
[Formula 3]
[184]

[185]
The compound of formula 2 (6-O-β-D-Psicopyranosyl-α-D-psicofuranose) is compound A, and the compound of formula 3 (6-O-β-D-Psicopyranosyl-β-D-psicofuranose) is compound B named as
[186]
On the other hand, D2 is a hydroxy group of carbon 2 (C2; according to carbon numbering in FIG. 4) of allulose in a structural isomer relationship with Formula 1 above, and any of carbons 1 to 6 (C1 to C6) of one molecule of allulose It was confirmed that they are novel allulose disaccharides, which are glycosidic bonds of a hydroxyl group of one carbon.
[187]
[188]
From the above description, those skilled in the art to which the present application pertains will understand that the present application may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present application should be construed as including all changes or modifications derived from the meaning and scope of the claims to be described later rather than the above detailed description and equivalent concepts thereof.
Claims
[Claim 1]
A composition for inhibiting HMF (Hydroxymethylfurfural) production, comprising allulose disaccharide.
[Claim 2]
The composition of claim 1, wherein the HMF production is due to saccharides.
[Claim 3]
The composition of claim 2, wherein the saccharide is a monosaccharide.
[Claim 4]
4. The composition of claim 3, wherein the monosaccharide is allulose.
[Claim 5]
The method of claim 1, wherein in the allulose disaccharide, two molecules of allulose are linked by a glycosidic bond, and the glycosidic bond is the second carbon (C2) of one molecule of allulose among the two molecules of allulose. The composition, wherein the hydroxyl group is a glycosidic bond to the hydroxyl group of any one carbon of the 1st to 6th (C1 to C6) carbons of one molecule of allulose.
[Claim 6]
A composition for preventing browning, comprising allulose disaccharide.
[Claim 7]
The composition of claim 6, wherein the browning is caused by saccharides.
[Claim 8]
The composition of claim 7, wherein the saccharide is a monosaccharide.
[Claim 9]
9. The composition of claim 8, wherein the monosaccharide is allulose.
[Claim 10]
7. The method of claim 6, wherein, in the allulose disaccharide, two molecules of allulose are linked by a glycosidic bond, and the glycosidic bond is the second carbon (C2) of one molecule of allulose among the two molecules of allulose. The composition, wherein the hydroxyl group is a glycosidic bond to the hydroxyl group of any one of carbons 1 to 6 (C1 to C6) of one molecule of allulose.
[Claim 11]
A method for inhibiting HMF production, comprising preparing a mixed composition comprising saccharides and allulose disaccharides.
[Claim 12]
A method for preventing browning, comprising preparing a mixed composition comprising saccharides and allulose disaccharides.
[Claim 13]
preparing a mixed composition comprising saccharides and allulose disaccharides; and heating the mixed composition.
[Claim 14]
The method of claim 13, wherein heating the mixed composition inhibits HMF production.
[Claim 15]
Use of allulose disaccharide to inhibit HMF production.