Beverage with suppressed growth of microorganisms

Abstract

A beverage containing one or more of the six kinds of compounds derived from natural products represented by Formulas 1 to 6 described in the specification or salts thereof. These compounds can exhibit an effect of suppressing the growth of microorganisms in the beverage at the same or lower concentrations than existing preservatives for beverages such as sodium benzoate and potassium sorbate. These compounds tend to be less likely to reduce the commercial value of the beverage.

Description

Technical Field

[0001]

[0001] The present invention relates to a beverage containing one or more specific compounds. More specifically, it relates to a beverage in which the growth of microorganisms is suppressed by containing one or more specific compounds derived from natural products. Even more specifically, it relates to a beverage in which the growth of microorganisms having the commercial value of a beverage product such as appearance stability is suppressed.

Background Art

[0002]

[0002] A preservative is an additive that suppresses the growth of microorganisms that cause spoilage and deterioration of food and beverages and enhances the shelf life. As a preservative for beverages, synthetic preservatives or synthetic antibacterial agents such as sodium benzoate and potassium sorbate are usually used. However, synthetic preservatives have recently been shown to affect the intestinal flora, and there are concerns about their effects on health. In addition, sodium benzoate is said to have a risk of generating carcinogenic benzene by reacting with vitamin C. In addition to the effects on health, sodium benzoate has a unique astringent taste (Patent Document 1) and may affect the flavor of beverages. In addition, potassium sorbate has a problem that its growth inhibitory effect on specific microorganisms is low and it has a unique odor.

[0003]

[0003] In order to reduce the use of such synthetic preservatives, studies have been conducted on preservatives derived from natural products (Patent Documents 2 and 3). Compounds derived from natural products such as chitosan are known to have antibacterial properties.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0005]

[0005] There are examples of studies on preservatives derived from natural products, but there are few examples where it has been confirmed that the growth of microorganisms is actually suppressed in beverages. Further, there are even fewer examples where it has been confirmed that the commercial value of beverages such as appearance stability and taste is not impaired. With the diversification of consumers' preferences in recent years, the types of beverage products have also diversified, and it is desirable to search for new natural product-derived preservatives suitable for use in beverages so as to be able to accommodate the various properties and flavors of various beverage products.

Means for Solving the Problems

[0006]

[0006] As a result of searching for natural product-derived compounds suitable for use as preservatives (microorganism growth inhibitors) for beverage products, the present inventors discovered six types of compounds. When tests were conducted to actually add these compounds to commercially available beverages and confirm their microorganism growth properties, it was found that these compounds have an effect of suppressing the growth of microorganisms. Further, it was found that these compounds have the appearance stability (property that the appearance hardly changes over time) required for beverage products and hardly affect the original color and / or taste of the beverages, leading to the present invention. More specifically, the six types of compounds are compounds represented by Formulas 1 to 6 described in the column of "Modes for Carrying Out the Invention" below.

[0007]

[0007] The present invention relates to a beverage containing any one or more of the compounds represented by Formulas 1 to 6 or a salt thereof. The present invention also relates to a beverage containing any one or more of the compounds represented by Formulas 1 to 6 or a salt thereof at a total concentration of 0.1 to 250 ppm. The present invention also relates to a beverage in which the growth of microorganisms is suppressed as compared with a beverage having the same composition except that it does not contain the compounds represented by Formulas 1 to 6 or a salt thereof when the microorganisms are added to the beverage and stored at room temperature for 2 weeks. The microorganisms are preferably yeast and / or mold, and more preferably any one or more of the genera Zygosaccharomyces, Saccharomyces, Brettanomyces, and Neosartorya. The present invention also relates to the above beverage in which the growth of microorganisms of the genus Alicyclobacillus is suppressed as compared with a beverage having the same composition except that it does not contain the compounds represented by Formulas 1 to 6 or a salt thereof. The present invention relates to a method for producing a beverage, which includes adding any one or more of the compounds represented by Formulas 1 to 6 or a salt thereof. The present invention also relates to a method for producing a beverage, which includes adding any one or more of the compounds represented by Formulas 1 to 6 or a salt thereof at a total concentration of 0.1 to 250 ppm. All of the compounds represented by Formulas 1 to 6 are compounds obtained from natural products, but as far as the inventors know, there has been no report to date of an example in which these compounds are actually added to beverage products to confirm the effect of suppressing the growth of microorganisms and the commercial value of the beverages.

Advantages of the Invention

[0008]

[0008] The beverage of the present invention contains any one or more of the compounds represented by Formulas 1 to 6 or salts thereof, thereby suppressing the growth of microorganisms in the beverage and having high preservability. All of the compounds represented by Formulas 1 to 6 are compounds derived from natural products. The beverage of the present invention containing any one or more of the compounds represented by Formulas 1 to 6 has the advantage that in beverages that had to use synthetic preservatives such as sodium benzoate and potassium sorbate, the growth inhibitory effect of microorganisms can be obtained without using these synthetic preservatives or by reducing the amount of these synthetic preservatives used. When using synthetic preservatives, the synthetic preservatives are usually added to beverages at about 200 to 400 ppm. However, the compounds represented by Formulas 1 to 6 or salts thereof have a growth inhibitory effect on microorganisms even at a concentration comparable to or lower than that of synthetic preservatives. One of the compounds represented by Formulas 1 to 6 or a salt thereof may be added to the beverage, or two or more of them may be combined and added to the beverage. The compound to be used may be selected according to the type and flavor of the beverage.

[0009]

[0009] The compounds represented by Formulas 1 to 6 have the advantage of not reducing the commercial value (for example, appearance stability, color, taste) of the beverage when added to the beverage. Chitosan is known, for example, as an antibacterial substance derived from natural products. However, chitosan is likely to become turbid over time depending on the type of beverage (that is, chitosan has somewhat low appearance stability). The compounds of Formulas 1 to 6 may exhibit appearance stability comparable to or higher than that when using chitosan at the same concentration when used at a concentration at which an antibacterial effect is obtained. In addition, when the compounds of Formulas 1 to 6 are used at a concentration at which an antibacterial effect is obtained, they are less likely to affect the original color of the beverage (less coloring property), and also tend to have little effect on the taste.

Embodiments for Carrying Out the Invention

[0010]

[0010] The present invention relates to a beverage containing any one or more of the compounds represented by Formulas 1 to 6 or salts thereof. Hereinafter, the compound represented by Formula 1 will be abbreviated as "Compound 1", and the compound represented by Formula 2 will be abbreviated as "Compound 2". Compounds 1 to 6 are all compounds obtained from natural products.

[0011]

[0011] Compound 1 (CAS No. 867578-35-6), Compound 2 (CAS No. 1427039-33-5), and Compound 3 (CAS No. 1427028-86-1) are compounds having the following Formulas 1 to 3, respectively, and are compounds extracted from the culture solution of Ustilago maydis, a kind of mold that grows on corn and the like.

[0012]

Chem.

[0013]

Chem.

[0014]

Chem.

[0015]

[0012] Compound 4 (C 51 H 82 O 21 )(CAS No. 68124-04-9) is a compound having the following Formula 4 and is a compound extracted from the roots of Dioscorea mexicana.

[0016]

Chem.

[0017]

[0013] Compound 5 (C 46 H 74 O 16 )(CAS No. 30994-75-3) is a compound having the following Formula 5 and is a compound extracted from the leaves of Securinega leucopyrus.

[0018]

Chem.

[0019]

[0014] Compound 6 (CAS No. 168778-19-6) is a compound having the following formula 6 and is a compound extracted from the roots of Agave americana.

[0020]

Chemical formula

[0021]

[0015] Compounds 1 to 6 can be obtained by extraction and purification from the above-mentioned microbial culture broth or plants. Alternatively, commercial products of each compound may be used and included in the beverage of the present invention. Or each compound may be obtained by synthesis by any method.

[0022]

[0016] Compounds 1 to 6 may be in the form of any salt in the beverage.

[0017] (Beverage) The beverage of the present invention contains any one or more of the above-mentioned compounds 1 to 6 or salts thereof. Depending on the type of beverage and the degree of the desired effect of inhibiting microbial growth, and considering the influence on the flavor of the beverage, one of the compounds 1 to 6 or salts thereof may be used, or two or more of them may be used in combination.

[0023]

[0018] The amount of each compound used in the beverage may be any amount as long as it can achieve the effect of inhibiting the growth of microorganisms in the beverage, and is not particularly limited. For example, as the lower limit, the total concentration of Compounds 1 to 6 or their salts is preferably 0.1 ppm or more, more preferably 0.5 ppm or more, further preferably 1.0 ppm or more, further preferably 10 ppm or more, further preferably 20 ppm or more, and further preferably 30 ppm or more. In order to minimize the impact on the flavor and appearance of the beverage, it is preferable to set the amount of Compounds 1 to 6 used in the beverage to be as small as possible within the range where the desired effect can be obtained. For example, as the upper limit, it is preferably 400 ppm or less, more preferably 300 ppm or less, further preferably 250 ppm or less, further preferably 125 ppm or less, further preferably 100 ppm or less, and further preferably 50 ppm or less.

[0024]

[0019] As described below, the beverage of the present invention may be a ready-to-drink beverage or a concentrated beverage for dilution and drinking. In the case of a concentrated beverage, in order to obtain the effect of inhibiting the growth of microorganisms in the concentrated state during storage, Compounds 1 to 6 or their salts that satisfy the above content may be used in the concentrated beverage. Alternatively, in order to obtain the effect of inhibiting the growth of microorganisms during storage in the diluted state of the concentrated beverage, according to the predetermined dilution ratio of the concentrated beverage, an amount of Compounds 1 to 6 or their salts that satisfies the above content in the diluted state is multiplied by the predetermined dilution ratio of the beverage, and the concentrated beverage may contain such an amount.

[0025]

[0020] Conventional synthetic preservatives such as sodium benzoate and potassium sorbate are usually used in beverages at a concentration of about 200 to 400 ppm. Compounds 1 to 6 or their salts may achieve the effect of inhibiting the growth of microorganisms in the beverage at a concentration comparable to or lower than that of conventional synthetic preservatives. In addition, Compounds 1 to 6 exhibit an effect of inhibiting the growth of microorganisms at a concentration comparable to or lower than that of chitosan, which is known as an antibacterial substance derived from natural products. Moreover, Compounds 1 to 6 are less likely to impair the commercial value of beverage products such as appearance stability.

[0026]

[0021] The method for measuring the content of each compound in the beverage is not particularly limited. For example, by preparing a standard sample of each compound and using HPLC or the like, the content can be measured by setting conditions according to the type of beverage.

[0027]

[0022] The type of beverage containing one or more of Compounds 1 to 6 or salts thereof is not particularly limited. For example, carbonated beverages, fruit juice beverages, vegetable juice beverages, coffee beverages, tea beverages, cocoa beverages, jelly beverages, milk-based beverages, probiotic beverages, soy milk, energy drinks, sports drinks, alcoholic beverages, and concentrated beverages to be diluted and consumed can be exemplified. Among them, acidic beverages with a pH of 2 to 5 (preferably pH 2.5 to 4.5) are preferred as beverages containing one or more of Compounds 1 to 6 or salts thereof because synthetic preservatives have been frequently used in these beverages. Examples of acidic beverages include carbonated beverages, fruit juice beverages, vegetable juice beverages, probiotic beverages, energy drinks, and sports drinks. The beverage may be a ready-to-drink beverage or a concentrated beverage for dilution and consumption. The type of fruit juice is not particularly limited, and examples include fruit juices of citrus fruits (lemon, grapefruit, lime, orange, mandarin orange, satsuma mandarin, tangor, natsumikan, amakatsu, hassaku, hyuga natsu, sudachi, yuzu, kabosu, daidai, iyokan, ponkan, kinkan, sanbokan, oro blanco, buntan), blackcurrant juice, apple juice, grape juice, peach juice, fruit juices of tropical fruits (pineapple, guava, banana, mango, acerola, papaya, passion fruit, lychee, etc.), ume juice, pear juice, apricot juice, plum juice, berry juice, kiwifruit juice, cherry juice, quince juice, watermelon juice, tomato juice, carrot juice, strawberry juice, and melon juice.

[0028]

[0023] The beverage of the present invention is preferably a beverage using any one or more of Compounds 1 to 6 or salts thereof in place of all or part of the synthetic preservatives conventionally used in beverages. Compounds 1 to 6 or salts thereof are preferably intentionally added for the purpose of enhancing the inhibitory effect on microbial growth in beverages separately from the normal components of the beverages.

[0029]

[0024] Normally, since beverages have diverse compositions, even substances with a generally known growth inhibitory effect on microorganisms cultured in a medium do not necessarily exhibit the same effect in beverages. For example, substances that are difficult to dissolve in beverages or substances that react with other components in beverages may not be able to inhibit the growth of microorganisms in beverages even if they are effective against microorganisms in the medium. Also, for example, pectin and polysaccharides contained in fruit juices and the like may reduce the antibacterial effect by encapsulating coexisting compounds. In contrast, in the examples of the present application described later, the growth inhibitory effects of Compounds 1 to 6 on various microorganisms were actually confirmed using existing carbonated beverages and fruit juice beverages.

[0030]

[0025] The beverage of the present invention is a beverage in which the growth of microorganisms is inhibited compared to a beverage (comparative beverage) having the same composition except for not containing Compounds 1 to 6 or salts thereof after adding microorganisms and storing at room temperature for 2 weeks. Here, the microorganisms are those that cause spoilage and decay of beverages, and examples include yeasts and molds. Typical examples of yeasts and molds that cause spoilage and decay include yeasts of the genus Zygosaccharomyces, Saccharomyces, or Brettanomyces, and molds of the genus Neosartorya. The beverage of the present invention is preferably one in which the growth of one or more of these microorganisms is inhibited compared to the above comparative beverage. Also, as microorganisms that cause spoilage and decay of beverages, thermophilic acidophilic bacteria (for example, the genus Alicyclobacillus) having high heat resistance in the strongly acidic range are known. Preferably, the beverage of the present invention is one in which the growth of thermophilic acidophilic bacteria is inhibited compared to the above comparative beverage.

[0031]

[0026] Compared with the comparative beverage, "the growth of microorganisms is suppressed" means that the growth of microorganisms that cause spoilage and decay of the beverage as described above is less than that of the comparative beverage. When microorganisms are added and stored at room temperature for two weeks, the fact that the growth of microorganisms is less than that of the comparative beverage indicates that the growth of the added microorganisms is suppressed due to the presence of Compounds 1 to 6 or salts thereof in the beverage. The procedure of "adding microorganisms and storing at room temperature for two weeks" is carried out for the purpose of testing the growth of microorganisms in the beverage of the present invention (microbial growth confirmation test). Of course, such addition of microorganisms is not performed on beverages shipped as normal products. The amount of microorganisms added in the microbial growth confirmation test is approximately 10,000 cfu / ml in the case of yeast and approximately 15 to 70 cfu / ml in the case of mold. Room temperature is 10 to 30 °C, preferably about 25 to 28 °C. Depending on the type of microorganism, it may be desirable to use a temperature other than room temperature for the test. Examples of specific methods for the microbial growth test include the methods described in the examples below.

[0032]

[0027] The beverage of the present invention is preferably a beverage in which, when microorganisms are added and stored at room temperature for two weeks, the growth of microorganisms is very small and thus no spoilage or decay is observed. More preferably, the beverage of the present invention is a beverage in which microorganisms do not grow under the above conditions.

[0033]

[0028] The beverage of the present invention also preferably has appearance stability to such an extent that the commercial value of the beverage is not impaired. Appearance stability means that the appearance of the beverage does not easily change over time. For example, it means that there is little change in the appearance of the beverage when the opened beverage is stored at room temperature for about two weeks. The beverage of the present invention also preferably has little change from the original color and taste of the beverage (the color and taste of a beverage having the same composition except for not containing Compounds 1 to 6 or salts thereof) while containing Compounds 1 to 6 or salts thereof.

[0034]

[0029] (Manufacturing method) The present invention also provides a method for manufacturing a beverage, which includes adding any one or more of Compounds 1 to 6 or salts thereof. The method of the present invention includes a step of adding any one or more of Compounds 1 to 6 or salts thereof during the normal manufacturing process of the beverage. The addition timing of each compound is not particularly limited, and it may be added at any stage of the beverage manufacturing process. For example, when adding and mixing normal beverage materials, Compounds 1 to 6 or salts thereof may be added and mixed, or after mixing the normal materials, Compounds 1 to 6 or salts thereof may be added and mixed. The Compounds 1 to 6 or salts thereof to be added are preferably obtained by extraction and purification from the above-described natural products (plant or microbial culture solution). Alternatively, commercially available products of each compound may be used. Further, each compound may be obtained by synthesis by any method. The Compounds 1 to 6 or salts thereof to be added may be in a solid form such as powder, or in a liquid form dissolved or suspended in a solvent. Also, the addition method of each compound is not particularly limited, and the addition and mixing methods of materials in the normal manufacturing of beverages may be used.

[0035]

[0030] The addition amount of Compounds 1 to 6 or salts thereof may be any amount as long as it can obtain an effect of suppressing the growth of microorganisms in the manufactured beverage, and is not particularly limited. For example, as a lower limit, as the total concentration of Compounds 1 to 6 or salts thereof, 0.1 ppm or more is preferable, 0.5 ppm or more is more preferable, 1.0 ppm or more is more preferable, 10 ppm or more is more preferable, 20 ppm or more is more preferable, and 30 ppm or more is more preferable. Also, as described above, as an upper limit, 400 ppm or less is preferable, 300 ppm or less is more preferable, 250 ppm or less is more preferable, 125 ppm or less is more preferable, 100 ppm or less is more preferable, and 50 ppm or less is more preferable.

[0036]

[0031] The type of beverage is not particularly limited. Examples of beverages are as described above. As described above, the above content may be adjusted according to whether it is a ready-to-drink beverage or a concentrated beverage, and in the case of a concentrated beverage, according to whether the effect of inhibiting microbial growth in the concentrated state or the effect of inhibiting microbial growth in the diluted state is intended.

Examples

[0037]

[0032] Hereinafter, the present invention will be described in detail with reference to experimental examples, but the present invention is not limited thereto. In this specification, unless otherwise specified, a numerical range is described as including its endpoints.

[0038]

[0033] (Yeast growth inhibition test 1) Using a commercially available carbonated beverage containing 1% lemon juice (containing saccharides (fructose, glucose syrup, granulated sugar), lemon juice, carbonic acid, flavor, vitamin C, acidulant, safflower pigment, calcium pantothenate, vitamin B6, carotene pigment) and a commercially available carbonated beverage containing 12% orange juice (fruits (orange, lemon, mandarin orange, grapefruit), saccharides (granulated sugar, fructose, glucose syrup), fruit fiber, orange peel extract, carbonic acid, flavor, antioxidant (vitamin C)), the growth inhibition effects of Compounds 1 to 6 and chitosan as a comparison on various yeasts in each beverage were confirmed. The specific procedure is as follows.

[0039] [

[0034] ]Each of the above beverages was uncapped, and the carbonated beverage was left standing until the carbon dioxide gas escaped, and then the pH of the beverage was adjusted to 3.3 using an aqueous sodium hydroxide solution. Each of Compounds 1-6 was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 20 mg / mL, and the resulting solution of each compound was added to the beverage so that the concentration of each compound in the beverage was 260.3 ppm. For comparison, an aqueous solution containing 20 mg / mL of chitosan and 20 mg / mL of ascorbic acid was prepared and added to the beverage so that the concentration of chitosan in the beverage was 260.3 ppm. A 96-well microplate was prepared, and 240 μL each of the beverage added with each of Compounds 1-6 and the beverage added with chitosan were dispensed into each well.

[0040] [

[0035] ]Bacterial solutions of 10,000 cfu / mL each of three types of yeasts (Zygosaccharomyces sp., Saccharomyces sp., Brettanomyces sp.) were prepared.

[0041] [

[0036] ]To the three wells added with the beverage containing Compounds 1-6 or chitosan, 10 μL each of the bacterial solution of each yeast was added (that is, three test sections of one well for each compound-yeast combination were prepared). Also, as a blank, 10 μL each of physiological saline was added to three of the wells added with the beverage containing Compounds 1-6 or chitosan. As a result, the final concentrations of Compounds 1-6 and chitosan in each well were 250 ppm. Also, as a compound-free section, a section was prepared by adding 10 μL each of the bacterial solution of each yeast to 240 μL of a beverage to which neither Compounds 1-6 nor chitosan was added.

[0042] [

[0037] ]The microplate prepared as described above was stored at 28°C for two weeks. After storage, after confirming that the yeast had grown in the compound-free section, in the beverage added with each compound, the growth of the yeast was visually confirmed in comparison with the blank. Specifically, the presence or absence of precipitation by the yeast was visually confirmed, and those in which precipitation was confirmed were considered to have growth. For each compound and each yeast, the number of wells in which the yeast grew out of the three wells is shown in Table 1.

[0043]

Table 1

[0044]

[0038] From the results in Table 1, it can be seen that Compounds 1 - 6, similar to chitosan whose antibacterial property is known, show the effect of suppressing the growth of yeast.

[0039] (Yeast growth test 2) Using Compounds 1 - 3, the growth of yeast was visually confirmed in the same manner as in Yeast Growth Test 1, except that the final concentration of each compound in each well was set to the concentration described in Table 2. For each compound and each yeast, the number of wells in which yeast grew out of 3 wells is shown in Table 2.

[0045]

Table 2

[0046]

[0040] From the results in Table 2, it can be seen that Compounds 1 - 3 show the effect of suppressing the growth of yeast at lower concentrations depending on the type of beverage.

[0041] (Mold growth test 1) Commercially available beverages containing 20% apple juice (containing apples, saccharides (fructose, glucose syrup, granulated sugar), acidulants, flavors) and 30% orange juice (containing fruits (oranges, mandarin oranges), saccharides (fructose, glucose syrup, granulated sugar), acidulants, flavors, vitamin C) were used. The growth inhibitory effect of Compounds 1 - 6, as well as chitosan and potassium sorbate for comparison, against mold in each beverage was confirmed. The specific procedure is as follows.

[0047]

[0042] The beverage was uncapped and the pH of the beverage was adjusted to 3.5 using an aqueous sodium hydroxide solution. Each of Compounds 1 to 6 was dissolved in DMSO to a concentration of 20 mg / mL, and the resulting solution of each compound was added to the beverage so that the concentration of each compound in the beverage was 250 ppm. Also, for comparison, an aqueous solution containing 20 mg / mL of chitosan and 20 mg / mL of ascorbic acid was prepared and added to the beverage so that the concentration of chitosan was 250 ppm. Similarly, for comparison, a beverage to which potassium sorbate was added so that the concentration thereof in the beverage was 250 ppm was also prepared. 3 mL each of the beverages to which each of Compounds 1 to 6 was added and the beverages to which chitosan or potassium sorbate was added were dispensed into centrifuge tubes with a volume of 15 mL.

[0048]

[0043] A spore suspension of mold (Neosartorya sp.) was added to each centrifuge tube so that the amount thereof was 50 to 200 cfu / tube.

[0044] Separately from the above, a blank without addition of the spore suspension was prepared. Also, as a compound-free group, a group was prepared in which a spore suspension of mold was added to 3 mL of a beverage to which none of Compounds 1 to 6, chitosan, and potassium sorbate was added so that the amount thereof was 50 to 200 cfu / tube.

[0049]

[0045] Each centrifuge tube prepared as described above was stored at 28°C for 8 weeks. After storage, after confirming that the growth of the mold mycelium was observed in the compound-free group, the growth of the mold was visually confirmed in the beverages to which each compound was added, in comparison with the blank. Specifically, the presence or absence of the elongation of the mold mycelium was visually confirmed, and those in which the elongation of the mycelium was confirmed were considered to have growth (+), and those in which no mycelium was confirmed were considered to have no growth (-). The results are shown in Table 3.

[0050]

Table 3

[0051]

[0046] Growth of mold was confirmed for chitosan when two weeks had passed at 28°C. Also, growth of mold was confirmed for potassium sorbate when four weeks had passed. In contrast, growth of mold was not confirmed even when eight weeks had passed at 28°C for Compounds 1 - 6. This result indicates that Compounds 1 - 6 have a higher mold growth inhibitory effect compared to chitosan, for which antibacterial properties are known, and potassium sorbate, which is an existing synthetic antibacterial agent.

[0052]

[0047] (Mold Growth Test 2) Growth of mold when eight weeks had passed at 28°C was visually confirmed in the same manner as in Mold Growth Test 1, except that the concentration added to each beverage was the concentration described in Table 4. The results are shown in Table 4.

[0053]

Table 4

[0054]

[0048] The results in Table 4 indicate that for some types of beverages, Compounds 1 - 6 have the effect of suppressing the growth of mold even when used at lower concentrations.

[0049] (Thermophilic Acidophilic Bacteria Growth Test) Using a commercially available beverage containing grape juice (grape, lemon juice), saccharides (fructose, granulated sugar), salt, lemon extract, lime extract, unshu mandarin extract, yuzu peel, dried tomato extract, flavoring, lactic acid, potassium chloride, antioxidant (vitamin C), the growth inhibitory effect of Compounds 1 - 6 and sodium benzoate as a comparison on thermophilic acidophilic bacteria in the beverage was confirmed. The specific procedure is as follows.

[0055]

[0050] The beverage was uncapped, and the pH of the beverage was adjusted to 3.3 using an aqueous sodium hydroxide solution. Each of Compounds 1 to 6 was dissolved in DMSO, and the solution of each obtained compound was added to the beverage so as to finally reach the concentrations shown in Table 5. As a comparison, a beverage to which sodium benzoate was added at the concentration shown in Table 5 was prepared. 3 mL each of the beverages to which each of Compounds 1 to 6 was added and the beverage to which sodium benzoate was added were dispensed into test tubes.

[0056]

[0051] A bacterial solution of Alicyclobacillus acidocaldarius at 400,000 cfu / mL was prepared.

[0052] To the test tubes added with the beverage containing Compounds 1 to 6 or sodium benzoate, 10 μL of the bacterial solution was added to each test tube. Also, as a non-additive compound group, a group in which the bacterial solution was added to a beverage to which neither Compounds 1 to 6 nor sodium benzoate was added was prepared.

[0057] Each test tube prepared as described above was stored at 45 °C for 2 weeks. After storage, 100 μL of the beverage was taken out from each test tube, smeared on YSGA medium, and then cultured at 55 °C for 3 days. After culturing, the number of colonies on the medium was counted. The number of bacteria was calculated based on the number of colonies. Those in which the number of bacteria increased by 10 times or more compared to the inoculated number of bacteria were considered to have growth (+), and those in which the increase in the number of bacteria from the inoculated number of bacteria was less than 10 times were considered to have no growth (-). The results are shown in Table 5.

[0058]

Table 5

[0059] As shown in Table 5, it was found that at least Compounds 1 to 3 and 5 exhibit a growth inhibitory effect against Alicyclobacillus acidocaldarius. The growth inhibitory effect of these compounds against Alicyclobacillus acidocaldarius is higher than that of sodium benzoate, an existing synthetic antibacterial agent.

[0060]

[0054] (Appearance Stability Test 1) The appearance stability of compounds 1 to 6 and chitosan for comparison was evaluated using the two types of commercially available beverages used in yeast growth test 1 and the two types of commercially available beverages used in mold growth test 1. The specific procedure is as follows.

[0061]

[0055] The above beverages were opened. The lemon carbonated beverage and the orange carbonated beverage were left to stand until the carbon dioxide gas escaped, and then the pH of the beverage was adjusted to 3.3 using an aqueous sodium hydroxide solution. Each of the compounds 1 to 6 was dissolved in DMSO at a concentration of 20 mg / mL, and the resulting solution of each compound was added to the beverage so that the concentration of each compound was 250 ppm. For comparison, a beverage was prepared to which an aqueous solution containing 20 mg / mL chitosan and 20 mg / mL ascorbic acid was added so that the concentration of chitosan in the beverage was 250 ppm. Each of the beverages to which the compounds 1 to 6 were added and the beverages to which the chitosan was added were dispensed in 240 μL portions into a 96-well microplate. The apple juice beverage and the orange juice beverage were adjusted to pH 3.5 using an aqueous sodium hydroxide solution. As in the case of the carbonated beverage, each of the compounds 1 to 6 was added to each beverage so that the concentration of each compound in the beverage was 250 ppm. For comparison, a beverage containing 250 ppm chitosan was prepared. 3 mL of each of the beverages containing compounds 1 to 6 and the beverage containing chitosan was dispensed into 15 mL centrifuge tubes.

[0062]

[0056] In addition to the above, a drink containing neither Compounds 1 to 6 nor chitosan was prepared as a compound-free group.

[0057] The microplates prepared as described above were stored at 28° C. for 2 weeks, and the centrifuge tubes were stored at 28° C. for 8 weeks. After storage, the appearances of the beverages containing the compounds 1 to 6 or chitosan were compared with the appearance of the beverages not containing any of the compounds, and scored according to the following criteria. 0 points: No difference in appearance compared to the compound-free area 1 point: There is a slight difference in appearance compared to the compound-free area. 2 points: There is a difference in appearance compared to the compound-free area. 3 points: The appearance is significantly different from that of the compound-free group, and the product has no commercial value as a beverage product.

[0058] In the above criteria, 0 to 2 points mean that the beverage product has the required quality (has beverage commercial value), and 3 points mean that the beverage product does not have the required quality (has no beverage commercial value). The results are shown in Table 6.

[0063]

Table 6

[0064]

[0059] The results in Table 6 show that Compounds 1 to 6 maintain the appearance stability that does not impair the commercial value of the beverage even when used at a concentration of 250 ppm depending on the type of beverage.

[0060] (Appearance Stability Test 2) Using Compounds 1 to 3 and lemon carbonated beverage and orange carbonated beverage, the appearance of the beverage was evaluated in the same manner as in Appearance Stability Test 1 except that the final concentration of each compound in each beverage was set to the concentration shown in Table 7. The results are shown in Table 7.

[0065]

Table 7

[0066]

[0061] As shown in Table 2, Compounds 1 to 3 exhibit an inhibitory effect on yeast growth against lemon carbonated beverage at a concentration of 31 ppm. The results in Table 7 show that Compounds 1 to 3 maintain the appearance stability that does not impair the commercial value of the beverage when used in lemon carbonated beverage at such a concentration.

[0067]

[0062] Also, as shown in Tables 1 and 2, Compounds 1 to 3 exhibit an inhibitory effect on yeast growth against orange carbonated beverage at a concentration of 31 ppm, and Compound 2 exhibits an inhibitory effect on yeast growth against orange carbonated beverage at a concentration of 250 ppm. The results in Tables 6 and 7 show that Compounds 1 to 3 maintain the appearance stability that does not impair the commercial value of the beverage when used in orange carbonated beverage at such a concentration.

[0068]

[0063] (Appearance Stability Test 3) Using Compounds 1 to 3, apple beverage, and orange beverage, the appearance of the beverages was evaluated in the same manner as in Appearance Stability Test 1, except that the final concentration of each compound in each beverage was set to the concentration shown in Table 8. The results are shown in Table 8.

[0069]

Table 8

[0070] As shown in Table 4, Compounds 1 to 3 exhibit an inhibitory effect on mold growth at a concentration of 31 ppm against apple beverage and orange beverage. The results in Table 8 mean that Compounds 1 to 3 maintain appearance stability that does not impair the commercial value of the beverages when used in apple beverage and orange beverage at such concentrations.

Claims

1. A beverage comprising any one or more of the compounds represented by the following formulas 1 to 6 or salts thereof: 【Chemical 1】 【Chemical 2】 【Chemical Formula 3】 【Chemical Formula 4】 【Chemical Formula 5】 [Chemical Formula 6]

2. The beverage according to claim 1, comprising any one or more of the compounds represented by formulas 1 to 6 or salts thereof at a total concentration of 0.1 to 250 ppm.

3. When stored at room temperature for two weeks after adding microorganisms, the growth of the microorganisms is suppressed as compared to a beverage having the same composition except that it does not contain the compound represented by formulas 1 to 6 or a salt thereof. The beverage according to claim 1 or 2.

4. The beverage according to claim 3, wherein the microorganism is yeast and / or mold.

5. The beverage according to claim 3, wherein the microorganism is any one or more of the genera Zygosaccharomyces, Saccharomyces, Brettanomyces, and Neosartorya.

6. The beverage according to claim 1 or 2, wherein the growth of microorganisms of the genus Alicyclobacillus is suppressed as compared to a beverage having the same composition except that it does not contain the compound represented by formulas 1 to 6 or a salt thereof.

7. A method for producing the beverage according to claim 1 or 2, comprising adding any one or more of the compounds represented by formulas 1 to 6 or salts thereof.

8. The method for producing a beverage according to claim 7, comprising adding any one or more of the compounds represented by formulas 1 to 6 or salts thereof at a total concentration of 0.1 to 250 ppm.

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