HK40004019A - Bacterial, fungal, and yeast growth inhibitor containing allulose - Google Patents

Description

Psicose-containing bacterial, fungal and yeast growth inhibitors

Technical Field

The present invention relates to a growth inhibitor for bacteria, fungi and yeast, which comprises a saccharide containing psicose.

Background

The fermented food is rich in viable microorganisms and is continuously fermented. As the time goes on, it is possible to,

the number of microorganisms in the fermented food is significantly changed, thereby causing quality changes such as changes in alcohol content or taste. Therefore, distribution and sale of fermented foods are restricted.

Specifically, unprocessed Magelleri (makgeolli), which is a fermented beverage

Has a short shelf life of 7 to 10 days, making it difficult to keep it on sale for a long period. In addition to this, the present invention is,

wine (wine), a fermented alcoholic beverage, cannot maintain quality due to after-fermentation, and therefore a sterilization process or a low-temperature storage facility is required to prevent such quality change from causing deterioration in organoleptic properties or increase in cost.

The present inventors have focused on inhibiting the post-fermentation of fermented alcoholic beverages. As a result, the present inventors found that addition of psicose can inhibit the growth of fungi (e.g., Aspergillus oryzae (Aspergillus oryzae), Aspergillus awamori (asp. awamori), Monascus purpureus (Monascus purpureus), Monascus ruber (Monascus ruber), and Rhizopus oryzae (Rhizopus oryzae)), bacteria (e.g., Lactobacillus casei and Lactococcus lactis subsp.

[ relevant documents ]

[ patent document ]

Korean patent No.10-1352025

Disclosure of Invention

Technical problem

An object of the present invention is to provide a growth inhibitor for microorganisms, which comprises a saccharide containing psicose.

It is another object of the present application to provide fermented alcoholic beverages comprising the growth inhibitory agent.

It is a further object of the present application to provide a method for inhibiting post-fermentation of a fermented alcoholic beverage, the method comprising adding the growth inhibitor to the fermented alcoholic beverage.

Technical scheme

According to one aspect of the present invention, a growth inhibitor for microorganisms comprises a saccharide containing psicose, wherein the microorganisms comprise at least one selected from the group consisting of the following bacteria, fungi and yeasts:

(i) lactobacillus casei and lactococcus lactis subspecies lactis as bacteria;

(ii) aspergillus oryzae, Aspergillus awamori, Aspergillus violaceus, Aspergillus rubrum, and Rhizopus oryzae as fungi; and

(iii) saccharomyces cerevisiae and Saccharomyces pastorianus as yeasts.

The psicose used herein may be directly extracted from a natural product, or may be chemically or biologically synthesized, but is not limited thereto. Further, the psicose may be provided in a crystal form or a syrup containing psicose (i.e., in a liquid form).

The content of the psicose may be 50 to 100 parts by weight with respect to 100 parts by weight of the saccharide based on the Dry Solid (DS) content. Specifically, the content of psicose may be 70 to 100 parts by weight, 90 to 100 parts by weight, 95 to 100 parts by weight, 98 to 100 parts by weight, 98.5 to 100 parts by weight, 99 to 100 parts by weight, or 99.5 to 100 parts by weight, relative to 100 parts by weight of the saccharide based on the dry solid content.

In addition to allulose, the saccharide may also comprise at least one sweetener. Examples of the sweetener may include any known sweetener (e.g., monosaccharide, disaccharide, oligosaccharide, sugar alcohol, and high intensity sweetener), but are not limited thereto. Examples of monosaccharides may include arabinose, xylose, fructose, tagatose, allose and galactose. Disaccharides refer to a group of carbohydrates consisting of two monosaccharide units linked together, and examples thereof may include lactose, maltose, trehalose, turanose, and cellobiose. Oligosaccharides refer to a group of carbohydrates consisting of 3 or more monosaccharide units linked together, and examples thereof may include fructooligosaccharides, isomaltooligosaccharides, xylooligosaccharides, gentiooligosaccharides, maltooligosaccharides, and galactooligosaccharides. Sugar alcohol refers to a compound obtained by reducing a carbonyl group in sugar, and examples thereof may include erythritol, xylitol, arabitol, mannitol, sorbitol, maltitol, and lactitol. The high intensity sweetener refers to a sweetener having a sweetness of ten times or more than that of sucrose, and examples thereof may include aspartame, acesulfame k, rebaudioside a and sucralose, but is not limited thereto. In another embodiment, the saccharide according to the invention may be free of sucrose, glucose or a combination thereof.

The growth inhibitor may inhibit the growth of lactobacillus casei or lactococcus lactis subspecies lactis such that the number of single cell colonies measured after 12 hours, 24 hours or 48 hours of culture in MRS medium at 37 ℃ is 300% or less, 200% or less, 180% or less, 150% or less, 130% or less, 120% or less or 100% or less of the initial number of single cell colonies cultured for 0 hours.

The growth inhibitor can inhibit the growth of aspergillus oryzae, aspergillus awamori, aspergillus kawachii, aspergillus rubrus, or rhizopus oryzae such that the number of single cell colonies measured after 24 hours, 48 hours, or 72 hours of culture in PD medium at 25 ℃ is 3,500% or less, 2,000% or less, 500% or less, 400% or less, 300% or less, 200% or less, 180% or less, 150% or less, 130% or less, 120% or less, 110% or less, or 100% or less of the initial number of single cell colonies cultured for 0 hour.

The growth inhibitor may inhibit growth of saccharomyces cerevisiae or saccharomyces pastorianus such that the number of single cell colonies measured after 24 hours, 48 hours, or 72 hours of culture in YM medium at 25 ℃ is 400% or less, 350% or less, 300% or less, 200% or less, 150% or less, 130% or less, 120% or less, 110% or less, or 100% or less of the initial number of single cell colonies cultured for 0 hour.

According to another aspect of the invention, a fermented alcoholic beverage comprises a growth inhibitor according to the invention.

The fermented alcoholic beverages may include any known fermented alcoholic beverages obtained by inoculating a fruit or grain with an alcohol-producing microorganism and then fermenting under specific conditions. Specifically, the fermented alcoholic beverage according to the present invention may be maggri wine, dongdong wine (dongdongju), low-alcohol rice wine (takju), wine or beer, more specifically, maggri wine or wine.

The fermented alcoholic beverage may be an alcoholic beverage fermented by at least one microorganism selected from the group consisting of the following bacteria, fungi and yeasts: (i) lactobacillus casei and lactococcus lactis subspecies lactis as bacteria; (ii) aspergillus oryzae, Aspergillus awamori, Aspergillus violaceus, Aspergillus rubrum, and Rhizopus oryzae as fungi; and (iii) Saccharomyces cerevisiae and Saccharomyces pastorianus as yeasts.

The fermented alcoholic beverage may be a fermented alcoholic beverage comprising at least one microorganism selected from the group consisting of the following bacteria, fungi and yeasts: (i) lactobacillus casei and lactococcus lactis subspecies lactis as bacteria; (ii) aspergillus oryzae, Aspergillus awamori, Aspergillus violaceus, Aspergillus rubrum, and Rhizopus oryzae as fungi; and (iii) Saccharomyces cerevisiae and Saccharomyces pastorianus as yeasts.

The fermented alcoholic beverage may further comprise food ingredients that are commonly used in fermented alcoholic beverages. In particular, the fermented alcoholic beverage according to the invention may further comprise at least one food ingredient selected from the group consisting of: purified water, oligosaccharides, rice, starch (e.g., maltodextrin), carbon dioxide gas, aspartame, organic acids (e.g., citric acid), and plant extracts.

In the fermented alcoholic beverages, the growth inhibitor, psicose, saccharides, bacteria, fungi and yeast are the same as described above.

According to yet another aspect of the present invention, a method for inhibiting post-fermentation of a fermented alcoholic beverage comprises adding a growth inhibitor according to the present invention to the fermented alcoholic beverage.

Embodiments of the present invention will now be described in more detail by a method of preparing a fermented alcoholic beverage. As described above, examples of fermented alcoholic beverages include representative Margri's wine and wine. First, embodiments of the present invention will be described in more detail by a method of manufacturing maogly wine, comprising the steps of:

(1) preparing a mother liquor by fermenting an unripe or cooked starch feedstock using bacteria, yeast or fungi;

(2) saccharifying the mother liquor prepared in step (1) by adding starch feedstock, saccharifying agent and water;

(3) fermenting the saccharified product obtained in the step (2), and then curing;

(4) removing undissolved starch solids by sieving the cooked product obtained in step (3); and

(5) preparing a final product by adding allulose, sugars and water to the sieved product obtained in step (4).

As in the above-described method of preparing magelline, since the method of inhibiting post-fermentation of fermented alcoholic beverages (such as wine) other than magelline according to the present invention is substantially the same as a typical wine production method except that saccharides and allulose according to the present invention are added instead of typical saccharides or sweeteners, a description thereof will be omitted.

In the method for suppressing the after-fermentation of fermented alcoholic beverages, the growth inhibitor, psicose, saccharides, bacteria, fungi and yeast are the same as described above.

Advantageous effects

According to the present invention, the growth of microorganisms in fermented alcoholic beverages can be suppressed, thereby suppressing the post-fermentation of fermented alcoholic beverages. Specifically, when allulose is used in place of sugars (e.g., sucrose) that are conventionally added to fermented alcoholic beverages (e.g., Margeli wine or wine)

The present invention can obtain the following effects: preventing unintentional quality deterioration that may occur during storage or distribution of fermented alcoholic beverages, and eliminating the need for a separate process or apparatus for improving storage stability, thereby simplifying the manufacturing process while reducing manufacturing costs.

Drawings

Fig. 1 and 2 are graphs showing time-dependent CFU values of bacterial targets (lactobacillus casei (fig. 1) and lactococcus lactis subspecies lactis (fig. 2)) as measured on samples of comparative examples using glucose or sucrose as a saccharide and examples using psicose as a saccharide.

Fig. 3 to 7 are graphs showing time-dependent CFU values of fungal targets (aspergillus oryzae (fig. 3), aspergillus awamori (fig. 4), aspergillus kawachii (fig. 5), aspergillus rubrus (fig. 6), and rhizopus oryzae (fig. 7)) as measured on samples of comparative examples using glucose or sucrose as a saccharide and examples using psicose as a saccharide.

Fig. 8 and 9 are graphs showing time-dependent CFU values of yeast targets (saccharomyces cerevisiae (fig. 8) and saccharomyces pastorianus (fig. 9)) as measured on samples of comparative examples using glucose or sucrose as a saccharide and examples using psicose as a saccharide.

Detailed Description

Hereinafter, the present invention will be described in more detail with reference to examples. It should be noted, however, that these examples are provided for illustration only and should not be construed as limiting the invention in any way. In addition, these examples are provided so that those of ordinary skill in the art will more fully understand the present invention.

Examples of the present invention

Experimental example 1: preparation of microorganisms and culture media for fermentation of fermented alcoholic beverages

In order to evaluate the inhibition of post-fermentation of fermented alcoholic beverages, the extent of inhibition of the growth of microorganisms commonly used in fermented alcoholic beverages was determined.

Specifically, each of the microorganisms (bacteria, fungi and yeast) obtained from the korean microbial culture center (KCCM) was subcultured three or four times in a culture medium (table 1) by a typical method to enhance activity. Then, the cultured microorganisms were inoculated into a medium (Table 1), and then were over-cultured to a concentration of 10⁵CFU/mL or more, thereby obtaining a strain mother liquor (Table 1).

TABLE 1

Experimental example 2: confirmation of microbial growth based on carbohydrate composition

In order to compare the degree of inhibition of microbial growth caused by the variation due to the kind of saccharides added alone, not due to the alcohol and other additives contained in the fermented alcoholic beverages, only the kind of saccharides added to each microbial culture medium was changed and studied.

Specifically, as sugars suitable for the medium of each strain, aqueous crystalline glucose (CJ cherijedang, purity 98% by weight or more), sucrose (CJ cherijedang, purity 98% by weight or more) and crystalline psicose (CJ cherijedang, purity 98% by weight or more) were added to each medium, thereby preparing modified media (tables 2 to 4). Then, 1mL of the mother liquor of each strain prepared in Experimental example 1 was inoculated into 100mL of the prepared modified medium, and then the time-dependent change in the number of cells of each strain was observed. That is, bacteria (lactobacillus casei and lactococcus lactis subspecies lactis), fungi (aspergillus oryzae, aspergillus awamori, aspergillus purpureus, aspergillus rubrus, and rhizopus oryzae), and yeasts (saccharomyces cerevisiae and saccharomyces pastorianus) were inoculated in the same manner in the MRS medium listed in table 2, the PDB medium listed in table 3, and the YMB medium listed in table 4, respectively. Then, each of the media inoculated with bacteria was stored at 37 ℃ for 12 hours, 24 hours, and 48 hours to be used as a sample for measuring the number of single cell colonies, and each of the media inoculated with fungi or yeast was stored at 25 ℃ for 24 hours, 48 hours, and 72 hours to be used as a sample for measuring the number of single cell colonies.

The measurement of the number of single cell colonies was performed by a typical microbiological assay. Specifically, 1mL of each sample was gradually diluted by a 10-fold dilution method using 9mL of 0.9% sterile physiological saline to obtain diluted samples. Then, 1mL of the diluted sample was placed in a petri dish, and 25mL of a standard medium supplemented with agar was poured into the petri dish so as to be sufficiently mixed and solidified, thereby obtaining an inoculated sample. Here, MRSA, PDA and YMA were used as standard media supplemented with agar for bacteria, fungi and yeast, respectively.

Thereafter, the number of measured cell colonies was multiplied by a dilution factor, thereby calculating a value of Colony Forming Units (CFU) per mL of sample.

TABLE 2

TABLE 3

TABLE 4

As a result, it was confirmed that the 9 strains of examples 1 to 3 using psicose as a saccharide had significantly lower cell colony numbers than the strains of comparative examples. Further, it was confirmed that the strains of comparative examples 1-1, 2-1 and 3-1 using glucose as a saccharide in a typical medium exhibited high growth of microorganisms, and the strains of comparative examples 1-2, 2-2 and 3-2 using sucrose as a saccharide exhibited growth of microorganisms without significant difference from the inoculated strain using glucose as a saccharide.

Specifically, in the medium using glucose as a saccharide (comparative example 1-1) and the medium using sucrose as a saccharide (comparative example 1-2), the cell numbers of Lactobacillus casei and lactococcus lactis as bacteria increased 40.6 to 137 times. In contrast, in the medium using psicose as a saccharide, the cell number of the bacteria was increased only 1.3-fold to 2.2-fold. Therefore, substantially no growth of the strain was observed (fig. 1 and 2).

Further, in the medium using glucose as a saccharide (comparative example 2-1) and the medium using sucrose as a saccharide (comparative example 2-2), the cell numbers of Aspergillus oryzae, Aspergillus awamori, Aspergillus kawachii, Aspergillus erythraea, and Rhizopus oryzae as fungi increased 9.8-fold to 148.5-fold and 5.9-fold to 81.1-fold, respectively. In contrast, in the medium using psicose as a saccharide, the cell number of the fungus was increased only 1.6-fold to 9.8-fold (example 2). Thus, a statistically significant inhibition of microbial growth was observed (fig. 3 to 7).

Finally, in the medium using glucose as a saccharide (comparative example 3-1) and the medium using sucrose as a saccharide (comparative example 3-2), the cell numbers of Saccharomyces cerevisiae and Saccharomyces pastorianus as yeasts increased 27.4-fold to 134.8-fold. In contrast, in the medium using psicose as a saccharide, the cell number of yeast was increased only 1.3 to 3 times. Therefore, substantially no growth of the strain was observed (fig. 8 and 9).

For each strain, the number of individual colonies was measured after 0 hour, 12 hours, 24 hours, 48 hours and 72 hours of culture in each medium. The results are shown in Table 5.

TABLE 5

In summary, it can be seen that when psicose is used in place of sucrose, which is commonly used as a sweetener added to fermented alcoholic beverages such as magelli, post-fermentation caused by microbial growth that may occur during the manufacture or circulation of fermented alcoholic beverages can be reduced.

Although a few exemplary embodiments have been described herein, it will be appreciated by those skilled in the art that these embodiments are given by way of illustration only, and that various modifications, changes, and alterations may be made without departing from the spirit and scope of the invention. Therefore, the embodiments and the drawings should not be construed as limiting the technical spirit of the present invention, but as illustrating the technical spirit of the present invention. The scope of the invention should be construed in accordance with the following appended claims so as to encompass all modifications and changes derived from the appended claims and equivalents thereof.

Claims (8)

1. A growth inhibitory agent for microorganisms comprising:

a saccharide containing psicose, a saccharide,

wherein the microorganism comprises at least one selected from the group consisting of:

(i) lactobacillus casei and lactococcus lactis subspecies lactis as bacteria;

(ii) aspergillus oryzae, Aspergillus awamori, Aspergillus violaceus, Aspergillus rubrum, and Rhizopus oryzae as fungi; and

(iii) saccharomyces cerevisiae and Saccharomyces pastorianus as yeasts.

2. The growth inhibitor according to claim 1, wherein the psicose is contained in an amount of 50 to 100 parts by weight relative to 100 parts by weight of the saccharide on a dry solid content basis.

3. The growth inhibitory agent of claim 1, wherein the saccharide is free of sucrose, glucose, or a combination thereof.

4. A fermented alcoholic beverage comprising the growth inhibitor according to any one of claims 1 to 3.

5. The fermented alcoholic beverage of claim 4, wherein the fermented alcoholic beverage is a Mageni wine or a wine.

6. The fermented alcoholic beverage of claim 4, wherein the fermented alcoholic beverage is fermented using at least one microorganism selected from the group consisting of:

(i) lactobacillus casei and lactococcus lactis subspecies lactis as bacteria;

(ii) aspergillus oryzae, Aspergillus awamori, Aspergillus violaceus, Aspergillus rubrum, and Rhizopus oryzae as fungi; and

(iii) saccharomyces cerevisiae and Saccharomyces pastorianus as yeasts.

7. The fermented alcoholic beverage of claim 4, wherein the fermented alcoholic beverage comprises at least one microorganism selected from the group consisting of:

(i) lactobacillus casei and lactococcus lactis subspecies lactis as bacteria;

(ii) aspergillus oryzae, Aspergillus awamori, Aspergillus violaceus, Aspergillus rubrum, and Rhizopus oryzae as fungi; and

(iii) saccharomyces cerevisiae and Saccharomyces pastorianus as yeasts.

8. A method of inhibiting post-fermentation of a fermented alcoholic beverage comprising: adding the growth inhibitor according to any one of claims 1 to 3 to the fermented alcoholic beverage.