JP2001178431A - Proliferation inhibitor against heat-resistant end spore- forming bacterium - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a proliferation inhibitor which is used against heat-resistant end spore-forming bacteria, has a high proliferation inhibiting action against the heat-resistant end spore-forming bacteria, has good biodegradability, and has excellent safety for human bodies. SOLUTION: This proliferation inhibitor against heat-resistant end spore- forming bacteria, characterized by containing as an active ingredient a fermentation liquid obtained by fermenting raw coffee beans.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION The present invention relates to the growth of heat-resistant spore-forming bacteria, which generally have no effective means other than heat sterilization methods under high temperature and high pressure conditions known as retort sterilization and chemical treatment. The present invention relates to a thermostable spore-forming bacterial growth inhibitor that can be suppressed and a method for producing the same.

[0002]

2. Description of the Related Art Conventionally, the biggest problem in maintaining the quality of processed foods using natural raw materials and materials is the control of microorganisms, and it is the manufacturer of the manufacturer that minimizes spoilage and deterioration caused by microorganisms and distributes them stably. Is required. In general, there are many microorganisms in nature and usually coexist with human beings, but their presence can sometimes be harmful. Thermostable spore-forming bacteria are known to have strong resistance to drugs and heat, and do not die under normal heat treatment conditions, but remain in the final product in the production stage and adversely affect quality. It is.

[0003] Heat sterilization, which is usually performed, has been used for a long time as a simple and effective means, and is still commonly used at present. However, the heat sterilization method has a remarkable adverse effect on the flavor, flavor, stability of useful components, and the like when the load and influence on the food material itself due to heating are considered,
Since there is a limit to the heating temperature and time from the balance with the bactericidal effect, a sufficient effect may not be expected for heat-resistant spore-forming bacteria represented by Bacillus, which is a resident bacterium.

[0004] Conventionally, in the food field, preservatives as food additives and shelf life improvers have been used for the purpose of preservation and fungicide.
It has greatly contributed to quality preservation during the preservation and distribution period of products such as processed foods and cosmetics (“Food Chemical 5 Preservatives Overview”, Food Chemistry Newspaper, May 2, 1993)
5th).

[0005] Under these circumstances, the importance of food preservation techniques has been increasing more and more than ever, and among them, the development of control techniques that have a remarkable effect on thermostable spore-forming bacteria has been desired. is the current situation. In particular, in the food field, it is essential to be safe in consideration of ingestion by the human body, but from such a viewpoint, preservatives mainly containing food additives such as sucrose fatty acid esters and glycerin fatty acid esters Many of the shelf life improvers are used ("Essential Emulsifiers, 2nd Edition", by Tohru Hidaka, ed., Eds., Published by Koshobo).

However, these preservatives and preservatives containing food additives as main agents have not been sufficiently satisfactory in terms of bactericidal efficacy against heat-resistant spore-forming bacteria.

[0007]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above-mentioned conventional problems and the present situation, and has been made in order to solve the problems. An object of the present invention is to provide a thermostable spore-forming bacterial growth inhibitor capable of exhibiting an effect.

[0008]

Means for Solving the Problems The present inventors have conducted intensive studies in order to solve the above-mentioned conventional problems and the like. As a result, fermented liquid obtained by fermenting green coffee beans has been added to heat-resistant spore-forming bacteria. They found a growth inhibitory effect and completed the present invention. That is, the present invention provides the following (1) and (2)
Exists. (1) A heat-resistant spore-forming bacterial growth inhibitor comprising, as an active ingredient, a fermented liquid obtained by fermenting green coffee beans. (2) The heat-resistant spore-forming bacterial growth inhibitor according to the above (1), wherein the fermented liquid contains a component represented by the following analysis data. Analytical data: Using an analytical ODS column (analytical ODS column manufactured by YMC, YMC-PACK ODS-AQ AQ-312, 6.0 × 150 mm)
Column temperature 30 ° C., flow rate 1.0 ml / min, methanol /
In high performance liquid chromatography using water / acetic acid (20/80 / 0.3) as an eluent, chlorogenic acid and caffeine, which do not exist in commercial coffee drinks but are detected under the same conditions, must be used. Contains two components that are eluted before.

The term "thermostable spore-forming bacterium" as defined in the present invention refers to a bacterium which forms a spore (spore) having heat resistance, and specifically, a bacterium belonging to the genus Bacillus (for example, subt.
ilis, cereus, stearothermophilus, megaterium, etc.), Clostridium bacteria (for example, Clostridium botulinum, spo
rpgenes, pertringens, etc.) and bacteria such as Actinomyces bacteria (eg, violaceus). The term “growth inhibition (effect)” for thermostable spore-forming bacteria as defined in the present invention refers to the increase or decrease in turbidity observed during the growth of microorganisms in a liquid medium, and the provision of a prescribed nutrient in accordance with ordinary methods. It refers to the difference in increase or decrease in the number of colonies that can be measured after a culture period after being poured into a petri dish together with a contained agar medium.

[0010]

Embodiments of the present invention will be described below in detail. The heat-resistant spore-forming bacterial growth inhibitor of the present invention is characterized in that a fermented liquid obtained by fermenting green coffee beans is used as an active ingredient.

The green coffee beans used in the present invention include, for example, Robusta Indonesian Robusta, etc., Riberica and Arabica mandelins, Mocamatari, Guatemala, and the like.
Kilimanjaro, Blue Mountain, Ethiopia Mocha,
It is not particularly limited as long as it is commonly used for coffee beverages, such as Hawaii Kona and Santos. However, these green coffee beans may be any commercially available as a raw material for drinking,
It is green beans before producing a coffee beverage, and excludes all roasted coffee beans in the present invention. In addition,
The above green coffee beans may be used alone or as a mixture of two or more.

In the present invention, when fermenting green coffee beans, nutrients can be fermented in the absence of nutrients, but preferably fermentation is performed in the coexistence of nutrients. The nutrient is not particularly limited as long as it can ferment green coffee beans efficiently. For example, 0 to 5% by mass (hereinafter, simply referred to as “%”) of glucose and soy peptone may be 0 to 5% by mass. 1%, dipotassium phosphate 0 to 0.5%, tryptone 0 to 2%,
A nutrient medium containing 0 to 1% of sodium chloride may be mentioned, and it varies depending on the type of green coffee bean. More preferably, glucose 5%, soypeptone 0.5%, dipotassium phosphate 0.1%, sodium chloride 1. Those containing 0% are desirable.

In the present invention, the amount of green coffee beans to be fermented when green coffee beans are fermented is 5 to 25%, preferably 10 to 20%, based on the total amount of green coffee beans.
It is. If the charged amount is less than 5%, the progress of fermentation is slow,
Since the amount of the obtained heat-resistant spore-forming bacteria inhibitor is small, the effect on the heat-resistant spore-forming bacteria is weak. In addition, when the charged amount exceeds 25%, the green coffee beans absorb moisture, so that the fermentation filtrate (insoluble from the fermentation solution) A solution obtained by separating and removing substances and the like is not sufficiently obtained, which is not preferable. In the present invention, the green coffee beans may be used as they are, or may be ground and used, and the ground may be ground to any particle size. The green coffee beans vary depending on the variety before fermentation. In that case, a heat treatment at a temperature of 60 ° C. or higher, preferably 60 to 70 ° C., for 10 to 60 minutes is preferable. Note that roasted coffee beans used for normal edible purposes are treated with boiling water according to boiling, which is different from the above-described heat treatment.

In the present invention, when fermenting green coffee beans, the pH is in the range of 4-6.5, preferably 5-5.
It is desirable to ferment in the range of ~ 6. Fermentation is initially p
The fermentation is started from H7, but when fermented to a pH of less than 4, the effect of inhibiting the growth of thermostable spore-forming bacteria is poor, and if the fermentation time is 6.5 or more, the effect may not be exhibited. Examples of the alkaline compound that adjusts the pH at the start of fermentation include, for example, carbonates or hydroxides of alkali metals such as sodium and potassium, specifically, sodium carbonate, monosodium phosphate, disodium phosphate,
Examples include potassium carbonate, sodium hydroxide, potassium hydroxide and the like. Preferably, those having a buffering capacity such as sodium carbonate, monosodium phosphate and disodium phosphate are desirable. The reaction temperature in the fermentation treatment is not particularly limited, but is usually 20 to 40 ° C, preferably 2 to 40 ° C.
5 to 37 ° C, and the fermentation time is usually 24 hours to 12 hours.
0 hours. Further, in the fermentation performed in the present invention, a bacterial strain to be inoculated and used is unnecessary, and the fermentation step is performed by a microorganism that naturally adheres to and remains on green coffee beans. For this fermentation, a simple open tank is sufficient, but any device used as a fermenter may be used, for example, commercially available from Shibata Scientific Instruments Co., Ltd.
An MDL-500 type bio-mentor can be used.

The heat-resistant spore-forming bacterial growth inhibitor of the present invention contains, as an active ingredient, a fermented liquid (solution) obtained by fermenting green coffee beans. It is preferable to use a green coffee bean fermentation filtrate from which microorganisms and the like have been removed. Examples of the method for obtaining a fermentation filtrate include, for example, a method using centrifugation, filter filtration using a sterilizing filter or the like, ultrafiltration, or the like, and any method capable of removing insoluble substances, microorganisms, and the like. May be used. The fermented liquid obtained from the fermentation of green coffee beans in the present invention refers to an analytical ODS column (analytical ODS column manufactured by YMC, YMC-PACK ODS-AQ AQ-3).
12,6.0 × 150 mm), high-speed liquid chromatography using methanol / water / acetic acid (20/80 / 0.3) as the eluent at a flow rate of 1.0 ml / min. It does not exist and means that it contains two components (a heat-resistant spore-forming bacterial growth inhibitor) that are always eluted before chlorogenic acid and caffeine detected under the same conditions.

FIG. 1 (a) is a characteristic diagram showing the results of high performance liquid chromatography of a commercially available coffee beverage.
(B) is a characteristic diagram showing the results of high performance liquid chromatography of a green coffee bean fermented liquid. Comparing FIG. 1 (a) and FIG. 1 (b), the fermented liquid of green coffee beans of (b) is not present in a commercial coffee beverage, but is always higher than chlorogenic acid and caffeine detected under the same conditions. It is evident that two components eluted earlier have been detected, and these components serve as thermostable spore-forming bacterial inhibitors as described below.

As described above, the heat-resistant spore-forming bacterial growth inhibitor of the present invention is obtained by using a fermented liquid obtained by fermenting green coffee beans in the absence or presence of nutrients using an active ingredient, preferably a sterile filter or the like. Fermented filtrate (aqueous solution) excluding insoluble substances and contaminants such as microorganisms is used as an active ingredient. It can be suitably used as a preservative or the like to be used. The amount of the heat-resistant spore-forming bacterial growth inhibitor of the present invention in processed foods and cosmetics varies depending on the use of processed foods such as soft drinks. 0.001%) to 5%. If the compounding amount of the heat-resistant spore-forming bacterial growth inhibitor is less than 10 ppm, the desired heat-resistant spore-forming bacterium growth inhibitory effect cannot be exhibited. Although the effect of inhibiting the growth of germinal spore-forming bacteria does not change, it may show unnecessary coloration and cause deterioration in quality.

The present invention also provides a method of fermenting green coffee beans in the absence or presence of nutrients. Preferably, the obtained fermented solution is further subjected to the use of a sterilizing filter or the like to remove insoluble substances and microorganisms. The resulting fermentation liquor, preferably a fermentation filtrate, is a heat-resistant spore-forming bacterial growth inhibitor that exhibits a growth-suppressing effect on heat-resistant spore-forming bacteria. .

The heat-resistant spore-forming bacterial growth inhibitor of the present invention thus constituted is obtained by fermenting green coffee beans, and the obtained heat-resistant spore-forming bacterial growth inhibitor is markedly resistant to heat. It exerts a growth inhibitory effect on sexually spore-forming bacteria. In the present invention,
Since raw coffee beans that are derived from natural products can be produced as a starting material, they are expected to have good biodegradability and high safety to the human body, and can be used by adding them to foods, soft drinks, etc. . Furthermore, in the present invention, it is possible to suppress the growth of bacterial spores that are difficult to control in the process of producing processed food, so that when used in the process of production, it is possible to provide stable production quality, and to use the heat-resistant spores of the present invention. In the case of processed foods and the like to which a forming bacteria growth inhibitor is added, it is not necessary to perform excessive heat sterilization, so that deterioration in quality due to heating can be suppressed as much as possible, and processed foods with a rich flavor can be provided. Even if it is added to a processed food having a useful component that is unstable with respect to, the growth of bacterial spores can be suppressed.

[0020]

EXAMPLES Next, according to test examples (Examples and Comparative Examples),
The present invention will be described in more detail, but the present invention is not limited to the following examples.

Test Example 1: Spore growth inhibitory effect test on heat-resistant spore-forming bacteria by fermented liquid of green coffee beans Bacillus was added to tryptic soy broth medium (manufactured by Eiken Chemical Co., Ltd.).
The bacterial spore concentration of the subtilis ATCC9372 strain spores was 1.75 ×
Inoculated and prepared by a conventional method so as to obtain 10 ³ cfu / ml, and fermented liquids of green coffee beans of Examples 1 and 2 shown below, and
A culture test tube to which 0.05% (1/2000 parts by mass) of Comparative Examples 1 and 2 are added is prepared. Using a small shaking culture device (Bio Photo Recorder TN-1506 manufactured by ADVANTECH) equipped with an optical system measurement function,
Shaking culture was performed while maintaining the environment at 5 ° C., and changes in turbidity over time (measurement wavelength: 660 nm) were measured. In the obtained microorganism growth curve, the lower the value of turbidity, the higher the spore growth inhibitory effect on thermostable spore-forming bacteria.
These results are shown in FIG.

(Example 1) An aqueous solution prepared by adding 10% of green coffee beans (Brazil) to sterilized water was prepared, and phosphoric acid was added using a nutrient medium (hereinafter, simply referred to as "nutrient source") shown in Table 1 below. PH with 1% aqueous solution of monosodium (Kanto Chemical)
Was adjusted to 7.0, and fermented at 25 ° C. for 24 hours. (Example 2) Raw coffee beans (manderin type) in sterile water
A 20% aqueous solution was prepared, and the pH was adjusted to 6.5 with a 1% aqueous solution of sodium carbonate (manufactured by Kanto Chemical Co., Ltd.) using the nutrients shown in Table 1 below, followed by fermentation at 32 ° C. for 30 hours. Was set to Example 2. (Comparative Example 1) Sucrose fatty acid ester (P-167 manufactured by Mitsubishi Chemical Corporation)
0) was taken as Comparative Example 1. (Comparative Example 2) Sucrose fatty acid ester (S-167 manufactured by Mitsubishi Chemical Corporation)
0) was designated as Comparative Example 2.

[0023]

[Table 1]

As shown in FIG. 2, the first embodiment according to the present invention
And 2 are Comparative Example 1 (P-1670) and Comparative Example 2 (S-1670)
As compared to Bacillus subtilis ATCC9372 strain spores,
It was found that a remarkable growth inhibitory effect was exhibited.

Test Example 2: Test of Spore Growth Inhibition Effect on Thermostable Spore-Forming Bacteria by Fermented Liquid of Coffee Beans Using the same bacterial spore medium solution and culture method as in Test Example 1 above, the following examples were used. 0.5% (1/200 parts by mass) of Comparative Examples 3 and 4 and Comparative Examples 3 and 4 were added, and the change in turbidity over time using the fermentation filtrate was measured in the same manner as in Test Example 1. These results are shown in FIG.

Example 3 An aqueous solution was prepared by adding 5% by weight of green coffee beans (Gatema seeds) to sterilized water, and potassium hydroxide (manufactured by Kanto Chemical Co., Ltd.) 1 was prepared using the nutrients shown in Table 1.
After adjusting the pH to 8.5 using a weight% aqueous solution, 25
What fermented at 50 degreeC for 50 hours was set to Example 3. (Example 4) Raw coffee beans (Santos species) 2 in sterile water
An aqueous solution containing 0% by weight was prepared, and 1% by weight of monosodium phosphate (manufactured by Kanto Chemical Co., Ltd.) was used using the nutrients shown in Table 1.
After adjusting the pH to 8.0 using an aqueous solution, the pH was adjusted to 2 at 25 ° C.
Example 3 was fermented for 0 hours. (Comparative Example 3) Sucrose fatty acid ester (P-167 manufactured by Mitsubishi Chemical Corporation)
0) was taken as Comparative Example 3. (Comparative Example 4) Sucrose fatty acid ester (S-167 manufactured by Mitsubishi Chemical Corporation)
0) is Comparative Example 4.

FIG. 3 shows a third embodiment of the present invention.
And 4 are Comparative Example 3 (P-1670) and Comparative Example 4 (S-1670)
As compared to Bacillus subtilis ATCC9372 strain spores,
It was found that a remarkable growth inhibitory effect was exhibited.

[Test Example 3: Spore growth inhibitory effect test in milk coffee beverage] Coffee beans (Kilimanjaro sp./
(Hawaii Kona seed; 60/40%) 75 g was added to 1 liter of hot water at 95 ° C., allowed to stand, extracted for 20 minutes, and then quenched with a plate cooler to 900 g of a coffee solution to obtain 10 g of milk.
0 g and granulated sugar (70 g) were added and dissolved by stirring to prepare milk coffee. Bacillus stearothermophilus ATC so that this milk coffee has 10 ³ per ml
C7953 strain spore solution was added, and the following Examples 5 and 6 and Comparative Examples 5 and 6 were added to 0.05% (1/20
After the addition and filling in a sterilized closed container, 55
The number of microorganisms derived from spores detected after storage at 3 ° C. for 3 months was evaluated by the number of colonies counted after culturing at 32 ° C. for 5 days after pour-out into a standard agar medium according to a conventional method. The results are shown in Table 2 below.

Example 5 An aqueous solution was prepared by adding 15% of green coffee beans (Gatema seeds) to sterilized water, and potassium hydroxide (manufactured by Kanto Kagaku Co., Ltd.) was prepared using the nutrients shown in Table 1 below.
After adjusting the pH to 7.0 using a 1% aqueous solution, 25 ° C.
Example 5 was obtained by fermenting for 24 hours. (Example 6) Raw coffee beans (Gatemala species) 2 in sterile water
An aqueous solution containing 0% was prepared, and the pH was adjusted to 7.5 using a 1% by weight aqueous solution of potassium carbonate (manufactured by Kanto Kagaku) using the nutrients shown in Table 1, and then fermented at 25 ° C. for 28 hours. This was used as Example 6. (Comparative Example 5) Sucrose fatty acid ester (P-167 manufactured by Mitsubishi Chemical Corporation)
0) was taken as Comparative Example 5. (Comparative Example 6) Sucrose fatty acid ester (S-167 manufactured by Mitsubishi Chemical Corporation)
0) is Comparative Example 6.

[0030]

[Table 2]

As is evident from the results in Table 2, the number of detected microorganisms was significantly lower in Examples 5 and 6 according to the present invention than in Comparative Examples 5 and 6.

Test Example 4: Test of Suppressing Spore Growth in Milk Coffee Beverage by Fermented Liquid of Heated Green Coffee Beans Coffee Beans (Kilimanjaro / Hawaii Kona; 60
Add 75 g to 1 liter of hot water at 95 ° C., extract for 20 minutes after standing, add 100 g of milk and 70 g of granulated sugar to 900 g of a coffee solution obtained by rapid cooling with a plate cooler, and stir to dissolve. Milk coffee was prepared. Bacillus stearothermophilus ATCC7953 strain spore solution was added to this milk coffee so that the number of Bacillus stearothermophilus ATCC7953 strain became 10 ³ cells / ml, and the following Examples 7 and 8 and Comparative Examples 7 and 8 were added to 0.05% (1/2000 mass). Part) was added and filled in a sterilized airtight container. After storage at 55 ° C. for 3 months, the number of microorganisms derived from spores detected was mixed with a standard agar medium according to a conventional method, and then cultured at 32 ° C. for 5 days. It was evaluated by the number of colonies counted later. The results are shown in Table 3 below.

Example 7 In sterile water, 6
An aqueous solution to which 15% by weight of green coffee beans (Gatemala sp.) Heated at 5 ° C. for 30 minutes was added, and 1% by weight of disodium phosphate (manufactured by Kanto Kagaku) using the nutrients shown in Table 1 below. % Was adjusted to 6.5 using an aqueous solution, and fermented at 25 ° C. for 41 hours. [Example 8] 10% by weight of green coffee beans (Gatemaara species) previously heated in sterile water at 70 ° C for 40 minutes in an aqueous solution
Was adjusted to pH 8.0 using a 1% by weight aqueous solution of sodium carbonate (manufactured by Kanto Kagaku) using the nutrients shown in Table 1 below, and then fermented at 25 ° C. for 25 hours. This was designated as Example 8. [Comparative Example 7] Sucrose fatty acid ester (P-167 manufactured by Mitsubishi Chemical Corporation)
0) is Comparative Example 7. [Comparative Example 8] Sucrose fatty acid ester (S-167 manufactured by Mitsubishi Chemical Corporation)
0) is Comparative Example 8.

[0034]

[Table 3]

As is evident from the results in Table 3, the number of microorganisms detected in Examples 7 and 8 according to the present invention was significantly lower than that in Comparative Examples 7 and 8.

[Test Example 5: Spore growth inhibitory effect test in boiled soybeans] 100 g of commercially available soybeans were put into 500 ml of tap water and cooked for 60 minutes. After the completion, the excess broth was discarded, and the temperature was returned to room temperature. Opening the plastic bag under sterile, and this soybean boiled beans 10 g, 10 ³ cells / 10
g of Bacillus stearothermophilus ATCC7953 strain spore fluid, and the following Examples 9 and 10,
And Comparative Examples 9 and 10 were 0.05% (1/2000).
After adding, mixing and filling, the mixture was stored in a constant temperature room at 55 ° C. for 3 months. The number of spore-derived microorganisms detected in each test was 32
The evaluation was performed based on the number of colonies counted after culturing at 5 ° C. for 5 days. The results are shown in Table 4 below.

[Example 9] In sterile water, 6
An aqueous solution containing 10% of green coffee beans (mandellin species) heated at 5 ° C. for 30 minutes was prepared, and disodium phosphate (manufactured by Kanto Chemical Co., Ltd.) 1 was prepared using the nutrients shown in Table 1 below.
After adjusting the pH to 6.0 using a weight% aqueous solution,
Example 9 was fermented at 44 ° C. for 44 hours. Example 10 Sterile water and 50 ° C., 35
An aqueous solution containing 20% by weight of green coffee beans (Gatema seeds) heated for 1 minute was prepared, and the pH was adjusted using a 1% aqueous solution of sodium carbonate (manufactured by Kanto Kagaku) using the nutrients shown in Table 1 below. After adjusting to 7.0 and fermenting at 32 ° C. for 20 hours, Example 10 was used. [Comparative Example 9] Sucrose fatty acid ester (P-167 manufactured by Mitsubishi Chemical Corporation)
0) is Comparative Example 7. [Comparative Example 10] Sucrose fatty acid ester (S-16 manufactured by Mitsubishi Chemical Corporation)
70) was designated as Comparative Example 8.

[0038]

[Table 4]

As is clear from the results in Table 4 above, Examples 9 and 10 according to the present invention correspond to Comparative Examples 9 and 10
It was found that the number of detected microorganisms was significantly lower than that of

Test Example 6: Spore growth inhibitory effect test in fresh tsubuan 100 g of commercially available red beans were placed in 500 ml of tap water, heat-treated for 30 minutes, and excess broth was discarded.
500 ml of fresh tap water was added again and cooked for 60 minutes. After completion, the excess broth was discarded, the temperature was returned to room temperature, and the red beans were crushed to obtain crushed beans. Opening the plastic bag under sterile, and this Tsubuan 10 g, sets the Bacillus stearothermophilus ATCC7953 strain spores solution such as a 10 ^3/10 g, further, Examples 11 and 12 below, as well as,
After adding Comparative Examples 11 and 12 in an amount of 0.05% (1/2000 parts by mass), mixing and filling, the mixture was placed in a 55 ° C.
Stored for months. The number of spore-derived microorganisms detected in each test was evaluated by the number of colonies counted after culturing at 32 ° C for 5 days after pour- ing into a standard agar medium according to a conventional method.
The results are shown in Table 5 below.

Example 11 An aqueous solution was prepared by adding 15% of green coffee beans (Santos species) previously heated in an aqueous solution to 65 ° C. for 30 minutes in sterile water, and the nutrient sources shown in Table 1 below were prepared. Disodium phosphate (Kanto Chemical) 1
% Was adjusted to 7.5 using an aqueous solution, and fermented at 25 ° C. for 26 hours. Example 12 In sterile water, an aqueous solution was previously prepared at 60 ° C., 35
An aqueous solution to which 16% of green coffee beans (Gatamara sp.) Heated for 1 minute was added was prepared, and the pH was adjusted using a 1% aqueous solution of sodium carbonate (manufactured by Kanto Chemical Co.) using the nutrients shown in Table 1.
Was adjusted to 8.5 and then fermented at 25 ° C. for 34 hours. [Comparative Example 11] Sucrose fatty acid ester (P-16 manufactured by Mitsubishi Chemical Corporation)
70) was set as Comparative Example 11. [Comparative Example 12] Sucrose fatty acid ester (S-16 manufactured by Mitsubishi Chemical Corporation)
70) was set as Comparative Example 12.

[0042]

[Table 5]

As is clear from the results shown in Table 5, the number of detected microorganisms in Examples 11 and 12 according to the present invention was significantly lower than those in Comparative Examples 11 and 12.

[Test Example 7: Test of Spore Growth Inhibition Effect in Fresh Tsubuan Using Coffee Fermented Extract Filtrate] 150 g of commercially available red beans were put into 500 ml of tap water, heated for 45 minutes, and excess broth was discarded. Thereafter, 500 ml of fresh tap water was added again and cooked for 60 minutes. After completion, the excess broth was discarded, the temperature was returned to room temperature, and the red beans were crushed to obtain crushed beans. Open the plastic bag under sterile conditions and
g and Bacillus stearothe to be 103 pieces / 10g
rmophilus ATCC7953 strain spores were inoculated, 0.03% (1/3333 parts by mass) of Examples 13 and 14 and Comparative Examples 13 and 14 shown below were added, mixed, filled, and filled at 55 ° C. It was stored in a constant temperature room for 3 months. The number of spore-derived microorganisms detected in each test was evaluated by the number of colonies counted after culturing at 32 ° C for 5 days after pour- ing into a standard agar medium according to a conventional method. The results are shown in Table 6 below.
Shown in

Example 13 An aqueous solution was prepared by adding 20% by weight of green coffee beans (manderin type) preliminarily heated in an aqueous solution to 60 ° C. for 45 minutes in sterile water, and the nutrients shown in Table 1 below were prepared. After adjusting the pH to 7.0 using a 1% by weight aqueous solution of disodium phosphate (manufactured by Kanto Chemical Co., Ltd.) using a source, fermentation was performed at 25 ° C. for 26 hours, and a sterilizing filter (Durapore membrane filter, Nippon Millipore) was used. 5μm
Next, Example 13 was filtered through a similar membrane filter of 0.22 μm. [Example 14] In sterile water, previously in an aqueous solution at 60 ° C, 35
An aqueous solution containing 20% by weight of green coffee beans (Gatamara sp.) Overheated for one minute was prepared, and the pH was adjusted using a 1% by weight aqueous solution of sodium carbonate (manufactured by Kanto Kagaku) using the nutrients shown in Table 1. After adjusting to 8.0, the mixture was fermented at 25 ° C. for 25 hours and filtered with a sterile filter (Durapore membrane filter, Nippon Millipore) 5 μm and then with a similar membrane filter of 0.22 μm to obtain Example 14. [Comparative Example 13] Sucrose fatty acid ester (P-16 manufactured by Mitsubishi Chemical Corporation)
70) was set as Comparative Example 13. [Comparative Example 14] Sucrose fatty acid ester (S-16 manufactured by Mitsubishi Chemical Corporation)
70) was set as Comparative Example 14.

[0046]

[Table 6]

As is clear from the results shown in Table 6, the number of detected microorganisms was significantly lower in Examples 13 and 14 according to the present invention than in Comparative Examples 13 and 14.

Comprehensively considering the results of the above Test Examples 1 to 7 (including FIGS. 2 and 3 and Tables 2 to 6), the fermented liquid or the fermented filtrate obtained by fermenting green coffee beans is used as the active ingredient. It was found that the thermostable spore-forming bacterial growth inhibitor exhibited a remarkable growth-suppressing effect on thermostable spore-forming bacteria.

[0049]

Industrial Applicability According to the present invention, a heat-resistant spore-forming bacterial growth inhibitor which has a high growth inhibitory activity against heat-resistant spore-forming bacteria, has good biodegradability, and is excellent in safety to the human body. Is provided.

[Brief description of the drawings]

1 (a) and 1 (b) are characteristic diagrams showing the results of high performance liquid chromatography between a commercially available coffee beverage and a fermented liquid coffee bean.

FIG. 2 is a characteristic diagram showing the effect of inhibiting the growth of thermostable spore-forming bacteria in Examples 1 and 2 and Comparative Examples 1 and 2.

FIG. 3 is a characteristic diagram showing the effect of inhibiting the growth of thermostable spore-forming bacteria in Examples 3 and 4 and Comparative Examples 3 and 4.

Claims (2)

[Claims] 1. A heat-resistant spore-forming bacterial growth inhibitor comprising, as an active ingredient, a fermented liquid obtained by fermenting green coffee beans. 2. The heat-resistant spore-forming bacterial growth inhibitor according to claim 1, wherein the fermented liquid contains a component represented by the following analysis data. Analytical data: Using an analytical ODS column (analytical ODS column manufactured by YMC, YMC-PACK ODS-AQ AQ-312, 6.0 × 150 mm)
Column temperature 30 ° C., flow rate 1.0 ml / min, methanol /
In high performance liquid chromatography using water / acetic acid (20/80 / 0.3) as an eluent, it was not present in commercially available coffee beverages, and was always present before chlorogenic acid and caffeine detected under the same conditions. Contain two components eluted in