JPS632577B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing a cariogenic food and drink, and more particularly to a method for producing a cariogenic food and drink using a novel sweetener that does not easily cause the formation of dextran on the tooth surface. The novel sweetener referred to herein is a sweetener containing an oligosaccharide in which 1 to 4 molecules of fructose are bound to the sucrose produced by the action of fructosyltransferase on sucrose. Hitherto, sucrose has been widely used in confectionery and foods, taking advantage of its superior properties such as good sweetness, body, and crystallinity. However, sucrose is a substrate for dextran eucrase produced by oral microorganisms. Therefore, continuous ingestion of sucrose produces a large amount of insoluble dextran in the mouth, promoting the formation of tooth moss, and is said to be a cause of tooth decay. The present inventors made use of the excellent properties of sucrose and, as a result of intensive studies on sucrose-related carbohydrates that are less likely to cause dental caries, discovered an oligosaccharide group obtained by the action of fructosyltransferase on sucrose. A substance in which one molecule of fructose is bound to sucrose (hereinafter referred to as GF ₂ ), a substance in which two molecules of fructose are bound to sucrose (hereinafter referred to as GF ₃ ), a substance in which three molecules of fructose are bound to sucrose (hereinafter referred to as (hereinafter referred to as GF ₄ ), a substance in which four molecules of fructose are bound to sucrose (hereinafter referred to as GF ₅ ), etc. are produced by Streptococcus mutans.
It has been found that not only is it not affected by the action of dextran eucrase produced by oral microorganisms, but it also has the effect of suppressing the production of insoluble dextran from sucrose by dextran eucrase. GF ₂ , GF ₃ , GF ₄ used here,
Oligosaccharides such as GF ₅ can be isolated and purified from a transferred sugar composition obtained by the action of fructosyltransferase on sucrose by carbon chromatography, ion exchange chromatography, etc., but this is not practical. It is preferable to use compositions of these oligosaccharides, and furthermore, cariogenic sugar alcohols such as sorbitol, mannitol, and maltitol, and artificial sweeteners such as dehydrochalcone and stevioside are added to such compositions. It can also be used. Further, after adjusting the pH of the sugar composition aqueous solution obtained by acting fructosyltransferase on sucrose to 7 to 9, nickel catalyst of 3 to 10% based on the solid matter was added, and the reaction temperature was 50 to 130°C. By performing catalytic reduction under reaction hydrogen pressure conditions of 50 to 120 Kg/cm ² and selectively catalytically reducing only glucose and fructose in the composition, it acts by converting these monosaccharides into sorbitol and mannitol. However, the sugar alcohol-containing composition obtained as a result of such treatment may contain, for example, sorbitol 37%, mannitol 2%
%, Seuucrose 10%, GF ₂ 22%, GF ₃ 22%,
Although it has a composition of 7% GF ₄ , such a composition does not produce insoluble dextran by oral microorganisms, and also produces little organic acid, making it a sweetener with a higher cariogenic effect ( (See Test Example 4). The transferred sugar composition obtained by acting fructosyltransferase on sucrose contains unreacted sucrose, oligosaccharides such as GF ₂ and GF ₃ produced by the transfer reaction, and glucose by-produced by the transfer reaction. etc. However, such transferred sugar compositions are also less susceptible to the action of dextran euclase of oral microorganisms, and as a result, the amount of insoluble dextran produced is small. This is because even if sucrose is present in the composition, oligosaccharides such as GF ₂ and GF ₃ suppress the production of insoluble dextran from sucrose, and insoluble dextran is produced from oligosaccharides such as GF ₂ and GF ₃ . This is due to not doing so. As described above, compositions containing oligosaccharides in which one to four molecules of fructose are bonded to sucrose produce a small amount of insoluble dextran, which itself is considered to be a major factor in the development of dental caries, as well as GF ₂ , Due to the fact that oligosaccharides such as GF ₃ themselves suppress the production of dextran from sucrose, they are resistant to caries, and their sweetness is 60 to 80 when sucrose is 100, and their sweetness is very good. Unique flavor and richness
have. Furthermore, since GF ₂ , GF ₃ , etc. are non-reducing sugars, they are less likely to be colored during processing. Furthermore, the viscosity and osmotic pressure of this sweetener are approximately the same as those of sucrose, and since it is non-crystalline, it can also suppress crystallization when mixed with sucrose, fructose, lactose, etc. Furthermore, the degree of freezing point depression of this sweetener is comparable to that of sucrose, and it also has excellent moisturizing properties. As mentioned above, this sweetener has various characteristics that conventional sweeteners are considered to have, so it can be used in place of conventionally used sweeteners such as sugar, starch syrup, and high-fructose corn syrup. It can be used in all foods and drinks, and the foods and drinks obtained in this way are cariogenic. Fructosyltransferase used in the production of the sweetener used in the present invention mainly acts on sucrose to cleave the β-1 and 2 bonds between fructose and glucose, and then transfers the fructose to sucrose. yields GF ₂ and further
It has the effect of transferring fructose to GF ₂ to generate GF ₃ . The reaction products are thus GF ₂ , GF ₃
Enzyme Nomenclature is an oligosaccharide in which fructose is bound to sucrose.
It is different from Inulosucrase [2.4.1.9.] and Levansucrase [2.4.1.10.] described in (Academic Press, 1978). This enzyme is a member of the Aspergillus genus (Aspergillus niger).
[The genus Aspergillus, Williams & Wilkins Corporation, 1965,
293 pages], Fusarium genus (Fusarium lini IAM5008, etc.),
Genus Gloeosporium (Gloeosporium KaKi)
IAM5011), etc.), Saccharomyces genus (Saccharomyces cerevisiae, etc.), Rhodotorulla genus (Rhodotorulla glutinis, etc.), Pichia genus (Pichia miso)
Oxygen derived from microorganisms such as Hansenula (Hansenula miso, etc.), Candida (Candida tropicalis, etc.), and oxygen derived from plants such as asparagus and Jerusalem artichoke. Enzyme is used. Fructosyltransferase of microbial origin is grown in a suitable medium such as 5.0% sucrose, 1.0% peptone, 0.7% meat extract,
A culture filtrate obtained by culturing each microorganism in a medium containing 0.3% NaCl at the optimum temperature of 25 to 30°C for 24 to 96 hours, and removing the bacterial cells by filtration or centrifugation after completion of the culture. Furthermore, an enzyme purified from this culture filtrate by a conventional enzyme purification method such as ultrafiltration, ammonium sulfate salting out, solvent precipitation, gel filtration, or ion exchange chromatography can be used. For plant-derived enzymes, the enzyme can be extracted after destroying the plant tissue by physical means such as grinding, and an extract thereof or an enzyme purified from the extract by a conventional method can be used. The desired cariogenic sweetener can be obtained by allowing the enzyme thus obtained to act on sucrose; however, as a result of various studies regarding industrial transfer reaction conditions, it is preferable to carry out the reaction under the following conditions. That is, the transfer reaction is carried out at a sucrose concentration of 5 to 70%, preferably 30 to 60%. In addition, the reaction pH and reaction temperature vary depending on the origin of the enzyme, but the pH is 4.0 to 7.0, the temperature is 25 to 65℃, preferably 50 to 60℃.
The desired transfer reaction can be carried out at ℃. The amount of enzyme used is 5 to 200 units, preferably 20 to 80 units per gram of sucrose. Here, the unit of enzyme is 1.0 ml of 5% sucrose solution, 1.0 ml of pH5.0 buffer solution, 0.5 ml of oxygen solution added, and reacted at 40℃ for 60 minutes. 1μ
The amount of enzyme that produces moles of glucose is expressed as one unit. After the rearrangement reaction is completed, the enzyme is deactivated by heating, decolorized with activated carbon, desalted with an ion exchange resin, and then concentrated to obtain the desired product. Analysis of the transition composition should be carried out by high performance liquid chromatography using, for example, a Microbondapak CH column (manufactured by Waters Limited) and a solvent system of acetonitrile:water (80:20 (v/v)). Can be done. The composition of the caries-resistant sweetener obtained in this way is, for example, 30% glucose, 11% sucrose,
%, GF ₂ 28%, GF ₃ 25%, GF ₄ 5%, GF ₅ 1
%, but the composition of each constituent sugar can take various values depending on the reaction conditions. The oligosaccharide GF ₂ is O-β-D-fructofuranosyl-(2→1)-O-β-fructofuranosyl-(2→1)-α-D-glucopyranoside, O
-β-D-fructofuranosyl-(2→6)-O-
β-glucopyranosyl-(1→2)-β-D-fructofuranoside, O-β-D-fructofuranosyl-(2→6)-O-β-fructofuranosyl-
(2→1)-α-D-glucopyranoside, etc.
GF ₃ is O-β-D-fructofuranosyl-
(2→[1-O-β-D-fructofuranosyl] ₂
→1)-α-D-glucopyranoside, O-β-D
-Fructofuranosyl-(2→6)-O[β-D-
Fructofuranosyl-(2→2)]-O-α-D-
Glucopyranosyl-(1→2)-β-D-fructofuranoside, etc., and _GF4 is O-β-D-
Fructofuranosyl-(2→[1-O-β-D-
It is estimated to be fructofuranosyl-2] ₃ →1)-α-D-glucopyranoside. Next, the new sweetener and its ingredient GF ₂ ,
The effects of GF ₃ , GF ₄ , and GF ₅ will be explained in detail using experimental examples. Using dextran eucrase obtained by culturing Streptococcus mutans ATCC 25175 strain,
Table 1 in Test Example 1 below compares the amount of insoluble dextran produced from GF ₂ , GF ₃ , GF ₄ and GF ₅ with that from sucrose. As is clear from the table, no insoluble dextran was produced from _GF2 , _GF3 , _GF4 , and _GF5 . Similarly, we investigated whether GF ₂ and GF ₃ suppressed the production of insoluble dextran from sucrose by dextran eucrase, and the results are shown in Table 2 in Test Example 2 below. As is clear from the table, it was found that both GF ₂ and GF ₃ suppressed the production of insoluble dextran from sucrose. In addition, sweeteners with different compositions were prepared under various transition conditions, and the amount of insoluble dextran produced from these compositions was compared with that of sucrose, as shown in Table 4 in Test Example 3 below. As can be seen from this table, the amount of insoluble dextran produced is lower in all compositions than in sucrose.
In particular, the content (wt%) of sucrose in the total solid weight of the sweetener composition is
The total content (wt%) of oligosaccharides such as GF ₂ , GF ₃ , GF ₄ , etc. is more than double, that is, the total content (wt%) of oligosaccharides relative to the sucrose content (wt%) in the total solid content. If the ratio is 2.0 or more,
The amount of insoluble dextran produced is
It is 50% or less, which is practically preferable. Test Example 1 Streptococcus mutans ATCC 25175 strain was cultured under anaerobic conditions in a medium containing glucose and tryptocase, and after removing bacterial cells, it was concentrated and purified by ultrafiltration to produce dextran. Euclase was prepared. Next, add 1.0ml of 1% sugar solution and 0.67M phosphate buffer (PH
7.0) Mix 1.5ml and 0.25ml of the above enzyme solution and incubate at 37°C.
After reacting for an hour, the water-insoluble dextran produced was centrifuged at 3000 rpm for 15 minutes to collect the precipitate, which was washed twice with 5 ml of 70% ethanol.
Dissolve in 2.5 ml of 1M KOH solution and add phenol
The amount of dextran produced was determined by the sulfuric acid method. In addition, sucrose, GF ₂ , GF ₃ ,
1% solutions of each of GF ₄ and GE ₅ were used. _GF2 ,
GF ₃ , GF ₄ , and GF ₅ are obtained from a transferred sugar composition obtained by the action of fructosyltransferase on sucrose, which is fractionated and purified by carbon chromatography, and then isolated to a single spot by thin layer chromatography. A fraction giving . Display the results -
Shown in 1.

[Table] As shown in Table-1, GF ₂ , GF ₃ , GF ₄ , GF ₅
No production of dextran was observed. Test Example 2 In the presence of sucrose using the dextran eucrase prepared in Test Example 1
It was investigated whether GF ₂ and GF ₃ inhibit the production of insoluble dextran from sucrose when GF ₂ and GF ₃ are added. The reaction conditions are sugar solution 1.0
ml (each containing carbohydrates listed in Table-2),
0.67M phosphate buffer (PH7.0) 1.5ml, oxygen solution 0.25ml
were mixed and reacted at 37° C. for 4 hours, and the amount of insoluble dextran produced in the reaction solution was determined in the same manner as in Test Example 1.

[Table] The numbers in parentheses in Table 2 indicate the amount of insoluble dextran produced from sucrose.
The index is shown when it is set to 100. Test Example 3 Sweeteners having the following composition were produced by reacting fructosyltransferase with sucrose under various conditions.

[Table] Seuculose (%)
When the amount of insoluble dextran produced from the above transferred sugar composition was measured in the same manner as in Test Example 1, the results shown in Table 4 were obtained.

[Table] Test Example 4 Streptococcus mutans Serotype C with maltose 0.27% L-cysteine hydrochloride 0.01%, L
- Glutamate sodium _salt 0.1%, _NH4HPO4
0.2%, _{MgSO4.7H2O 0.02} %, NaCl 0.001%,
After culturing under anaerobic conditions in a medium containing 0.01% MnSO ₄ and 0.01% FeSO ₄ 7H ₂ O, the cells were collected by centrifugation and placed in 0.05M phosphate buffer for 10 min.
Disperse to give mg/ml. The amount of lactic acid produced is
0.2M-phosphoric acid buffer 0.9ml, 22.5M-MgCl ₂ 0.4
ml, 0.2ml of 1.7% sugar solution, and 0.5ml of bacterial cell dispersion,
After performing a shaking reaction at 37°C for 30 minutes, the reaction was stopped by boiling for 15 minutes, and after removing bacterial cells by centrifugation, the amount of lactic acid in the supernatant was determined by an oxygen method.

[Table] Next, examples will be described in which this sweetener was applied to the production of various foods and drinks. Example 1 Production of hard candy A non-cariogenic sweetener (moisture 25% W/W) was concentrated in vacuum to a final moisture content of 8 to 10%, and after cooling to about 80°C,
Add fragrance and coloring, mold and cool to room temperature.
Got a hard demand. Unlike candy made using sugar, this candy did not have the so-called "recovery" caused by the formation of sugar crystals, and was resistant to caries. Example 2 Manufacture of orange marmalade Soaked in 3% saline overnight to remove bitterness, washed with water to remove salt, and then added 90g of orange peel, which had been boiled for about 20 minutes to soften the tissue, and 200g of rainbow yeast. After adding 370 ml of water to this, it was boiled for about 30 minutes while adding 320 g of a caries-resistant sweetener (moisture 25% W/W). The sugar content was 65% w/w. This orange marmalade had a good flavor with the refreshing acidity of the orange. Example 3 Production of Neriyokan Soak 12g of agar in water for 3 hours, then drain the water.
This was crushed. Next, 260 ml of water was added and dissolved by heating. This is a non-cariogenic sweetener (moisture 25%)
Add 960 g of W/W) and filter after the agar is completely dissolved. Heat this agar and add 500 g of fresh bean paste.
After adding and mixing and boiling down to a sugar content of 70-71%,
I tried to put it in a sink and let it harden. This is a non-carious yokan. Example 4 Production of ice cream 10 parts skim milk, 75.5 parts water, 0.25 part stabilizer, emulsifier
A mix was prepared by adding an appropriate amount of flavoring to 0.25 parts of a cariogenic sweetener and 14 parts of a caries-resistant sweetener, which was filtered and then sterilized at 70°C for 30 minutes. This was then cooled. Temperature 3~
After aging at 5° C. for 6 hours, the temperature was lowered and frozen while stirring to produce ice cream. Since the degree of freezing point depression of the non-cariogenic sweetener is about the same as that of sugar, ice cream with good shape retention was obtained in the same way as when sugar was used. Example 5 Manufacture of biscuits 1 kg of wheat flour, 100 g of corn starch, 333 g of anti-cariogenic sweetener (moisture 25% W/W), 125 g of margarine
g, salt 5g, soda carbonate 2.5g, ammonium carbonate
A dough was prepared using 8.8 g of soybean lecithin, 6.3 g of soybean lecithin, 75 g of whole eggs, 6.3 g of vanilla oil, and 267 g of water, and after being rolled out, the dough was molded and roasted to produce biscuits.
The condition of the dough was the same as when sugar was used, and the volume and baked color of the product were also good. Example 6 Production of soft drink 1.5 parts of citric acid and 970 parts of water were added and dissolved in 133 parts of a non-carious sweetener (moisture 25% W/W). Nation was conducted to produce soft drinks. This is a non-cariogenic soft drink as it contains only a non-cariogenic sweetener. Example 7 Manufacture of chewing gum After dissolving and mixing 75 parts of caries-resistant sweetener powder and 22 parts of gum tickle, flavor and menthol were added and mixed, kneaded, rolled to a certain thickness with a pressing roll, and then cut. , and dried to produce a gum board. This gum uses only a non-cariogenic sweetener, so it is a non-cariogenic chewing gum. Example 8 Production of Chiyocolate Thiyocolate was produced by a conventional method by blending 100 parts of bitter chiyocolate, 116 parts of cariogenic sweetener powder, 23 parts of cocoa butter, 90 parts of powdered milk, vanilla, and a small amount of lecithin. The sweetness of this Chiyokolate was elegant, and the taste was no different from products made with sugar. Example 9 Production of Tsukudani Soy sauce 1, anti-cariogenic sweetener (moisture 25% W/W)
After mixing 900g and boiling to concentrate, 800g of clam meat
50g of ginger and 50g of ginger were added and boiled for 40 to 60 minutes to obtain boiled clams. The color and taste of the product were good. Example 10 Manufacture of marron glace After removing the outer skin of chestnuts and boiling them for 8 to 10 hours, the astringent skin was removed and a hot sugar solution containing a 40% non-cariogenic sweetener was poured onto the chestnuts.After leaving the chestnuts for a day, the concentration of the sugar solution was reduced. Pour a 45% solution and leave it for a day. In this way, the concentration of the sugar solution was increased to 70% to produce marron glacée. Since the osmotic pressure of the non-cariogenic sweetener is almost the same as that of sugar, a good product was obtained.

Claims (1)

[Scope of Claims] 1. A method for producing a caries-resistant food and drink, characterized in that a sugar mixture whose main component is an oligosaccharide in which 1 to 4 molecules of fructose are bound to sucrose is used as a sweetener. 2. A composition in which the ratio of the total content (wt%) of oligosaccharides in which 1 to 4 molecules of fructose are bonded to sucrose to the content (wt%) of sucrose in the total solid content is 2.0 or more as a sugar mixture. 2. A method according to claim 1, which uses a product. 3. The method according to claim 1, wherein an oligosaccharide in which 1 to 4 molecules of fructose are bonded to sucrose is used as the sugar mixture.