KR100780030B1 - Method for manufacturing soybean fermented beverage using vegetable lactic acid bacteria or Bacillus bacteria - Google Patents

Abstract

The present invention relates to a method for producing a soybean fermented beverage, comprising the steps of: (a) preparing a medium comprising a soybean pulverized liquid obtained by milling soybeans; (b) inoculating the medium with vegetable lactic acid bacteria, plant Bacillus bacteria or mixed bacteria thereof; And (c) culturing the bacteria inoculated in the medium to obtain a soybean fermented beverage.

Soybean, pretreatment, vegetable lactic acid bacteria, Bacillus bacteria, culture, fermented beverage

Description

Method for preparing fermented bean beverage using plant lactic acid bacteria or Bacillus bacterium {Method for Preparing Fermented Bean Beverage Using Plant-Originated Lactic Acid Bacteria or Bacillus}

Figure 1 is a schematic diagram showing an embodiment of the present invention for producing a soybean fermented beverage as a raw material of the method of the present invention.

The present invention relates to a method for producing a soybean fermented beverage, and more particularly, to a method for producing a soybean fermented beverage having a distinctive flavor and various physiological activities by inoculating and culturing the soybeans pretreated with vegetable lactic acid bacteria and / or Bacillus. It is about.

Lactic acid bacteria (LAB) is an industrially important microorganism that is widely applied worldwide for the production of various fermented food products, including fermented dairy products such as cheese, fermented milk and lactic acid bacteria drinks. In particular, most of the fermented milk in Korea can be said to be a representative food prepared to facilitate the intake of live lactic acid bacteria prepared using milk as a raw material.

Lactobacillus taken from fermented milk promotes the growth of intestinal useful bacteria, while improving the balance of the intestinal flora so that beneficial bacteria take the lead by inhibiting the growth of harmful bacteria. As a result, ① Intestinal infection and food poisoning prevention, ② Intestinal corruption suppression It has been shown to have various physiological functions such as preventing constipation, increasing immune system, inhibiting carcinogenesis, and producing vitamin B groups. Therefore, in the West, such as Europe, yogurt, lactic acid bacteria beverages and probiotic products in the form of powders, granules, and tablets form the largest market as functional foods.

In the case of fermented milk having such a function, most fermented strains are imported from foreign countries. In particular, these lactic acid bacteria are strains isolated from fermented dairy products such as cheese. Therefore, in the case of general lactic acid bacteria, it is true that the fermentation of products other than milk has limitations on the flavor of fermented products and various bioactive substances produced during fermentation.

Recently, apart from the lactic acid bacteria isolated from milk, cheese, etc., the advantages of vegetable lactic acid bacteria have been highlighted. Plant Lactobacillus is called Plant Originated Lactic Acid Bacteria (POLAB) and is treated differently from general lactic acid bacteria and refers to lactic acid bacteria isolated from fermented foods of plant raw materials that have been ingested since ancient times. In other words, the Kimchi of Korea is a representative report of the plant lactic acid bacteria (POLAB). These plant lactic acid bacteria are known to be excellent in the production of various bioactive substances because they grow in a harsh environment than the general lactic acid bacteria.

Soy is a protein and fat rich food that has been used in our diet. In general, there are various types of soybeans, soybeans, green beans, black beans, Seomoktae, peas, and kidney beans, which are consumed as processed products such as tofu and soy milk, or when they are cooked or cooked rice flour according to their characteristics. Sometimes used.

Soybeans have a slight difference in their function and composition, but 40% of the soybean protein contains essential amino acids in a balanced proportion, and other food proteins contain lysine which is easy to lack. to be. The fat content is about 18%, mostly unsaturated fatty acids, and more than 50% is the finest linolenic acid. In addition, vitamin E, which helps the action of linolenic acid, is enough to remove cholesterol caused by excessive intake of animal fat. In addition, it is rich in vitamin B, which helps to recover from fatigue, and it is also rich in calcium and iron. In addition, the food value of soybeans, such as anticancer action, atherosclerosis and anti-aging, is incredible.

Soybeans play an important role as a nutritional source of protein and fat, but their tissues are hard to absorb and digest. In addition, physiologically inhibited substances such as carbohydrates and trypsin inhibitors that are poorly digested may cause problems with absorption of digestion and cause side effects such as diarrhea.

When fermentation using microorganisms that can be used for food with soybean as raw material, it can solve such problems of digestion absorption and flavor by various enzymes produced by microorganisms during fermentation. It is possible to add the function of various bioactive substances produced.

Fermented beverages made from soybeans, which have been recently produced in Korea, are products that use lactic acid bacteria to produce fermented milk. Since soybean is a vegetable raw material, it is advantageous to enhance the flavor and function of vegetable lactic acid bacteria rather than general lactic acid bacteria for fermentation.

Also, doenjang, a pronoun of fermented foods except Kimchi, is a report of various Bacillus bacteria. As is widely known, Bacillus bacteria fermenting soybean paste are known to produce various bioactive substances such as thrombolytic enzymes.

When looking at a beverage-related patent that uses soybeans as a raw material applied or registered in Korea, raw materials are not only limited to soybeans, but also soybean powder prepared by extracting only water-soluble components of soybean (Patent No. 0348461), or soymilk. Was used (Patent No. 0471631). This is because the first processed soybean is removed from the rich dietary fiber contained in soybean hull and bean curd to reduce the physiological effect of the final product and has the disadvantage that some of the useful components of the soybean is removed. In addition, the strains used in the fermentation products using soybean powder and soybean oil, Bifidobacterium brave (Patent No. 0471631), Lactobacillus vulgaris, Lactobacillus casei and Lactobacillus Ashdophyllus mixed strains or one of them As the (Patent No. 04547620), lactic acid bacteria used for fermentation of dairy products other than vegetable lactic acid bacteria were mainly used. Soybean was fermented using Bacillus pilmus (Patent Application No. 2005-0023518), or Bacillus subtilis (Patent Application No. 2001-0069777), respectively, or using a two-stage fermentation method, By using a solid fermentation type rather than a beverage can be said that there are many differences in terms of configuration and function to seek in the present invention.

Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

The present inventors have diligently researched to establish a production system of soybean fermented beverages improved from the prior art, and as a result, fermented with vegetable lactic acid bacteria and / or vegetable Bacillus bacteria as a fermentation raw material as a whole part of soybean, the taste and nutrition By confirming that the fermentation beverage can be produced excellent in both aspects, the present invention was completed.

Accordingly, an object of the present invention is to provide a method for producing a soybean fermented beverage using plant lactic acid bacteria and / or Bacillus bacteria.

Another object of the present invention to provide a soybean fermented beverage comprising a vegetable lactic acid bacteria and / or Bacillus bacteria.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

According to one aspect of the invention, the present invention provides a method for producing a soybean fermented beverage, comprising the steps of: (a) preparing a medium comprising a soybean pulverized liquid obtained by grinding the beans; (b) inoculating the medium with vegetable lactic acid bacteria, plant Bacillus bacteria or mixed bacteria thereof; And (c) culturing the bacteria inoculated in the medium to obtain a soybean fermented beverage.

According to another aspect of the present invention, the present invention is a soybean fermented beverage prepared by the method of the present invention, the soybean fermented beverage is soybean fermentation, characterized in that it comprises vegetable lactic acid bacteria, vegetable Bacillus bacteria or mixed bacteria thereof Serve a drink.

The present inventors first selected plant originated Lactic Acid Bacteria (POLAB) and / or plant Bacillus bacteria as fermentation strains, and also to pre-process various soybeans in order to establish a production system of soybean fermented beverages improved from the prior art. The best protocol for producing soybean fermented beverages was established. According to the method of the present invention, through the minimum heat treatment and enzyme treatment can be used as a fermentation raw material without removing the whole bean, and to remove the soybeans to provide a soybean fermentation beverage with excellent taste and rich nutritional ingredients can do.

Described in each step the manufacturing method of soybean fermented beverage of the present invention as follows:

(a) preparation of soybean medium

First, a medium containing a soy flour obtained by milling soybeans is prepared.

In the present invention, it is advantageous to use the same type of beans continuously in the same region so that the final soybean fermented beverage maintains the same quality, but the difference of the finished product according to the quality difference is a slight difference in flavor and the fermentation itself. It doesn't matter much. In particular, there are some differences in nutritional components depending on the type of soybean, but the type of soybean does not have a significant effect on the fermentation itself because the difference in carbohydrate and protein content does not affect fermentation. In addition, the use of imported soybeans with the same nutritional composition does not significantly affect the quality of the product. According to a preferred embodiment of the present invention, the beans used as raw materials in the present invention are soybeans, green beans, black beans, Seomoktae, peas or kidney beans.

Before grinding the beans, it is preferable to first go through the washing process. Soybeans are usually encased in a sheath, so there is very little pesticide left on the surface, so excessive cleaning is unnecessary. It is preferable to use cold water of 30 ° C. or less, and the washing process is advantageously treated in the shortest time possible within 30 minutes in order to prevent leakage of the water-soluble nutrient. If the wash water temperature is over 30 ℃ and the washing time is over 30 minutes, the nutrition of soybean may be lost.

The washed beans are preferably called water. The process called water is usually carried out up to the maximum absorption of soybeans. The process is also good to proceed at a temperature of less than 30 ℃ If the water temperature is higher than this, there is a problem in that the growth of other microorganisms in the summer is possible during the process. The time required is 4-5 hours in summer (25-32 ° C) and 16-24 hours in winter (5-15 ° C).

This is crushed using beans soaked in water. Grinding of the beans can be carried out according to a process commonly known in the art, preferably using a fine grinding machine. The final particle size of the milled product is preferably 20-200 μm, more preferably 30-150 μm. When the particle size of the pulverized product exceeds 200 μm, there is a problem in that the texture is poor after fermentation, and when the particle size is less than 20 μm, a large amount of energy is consumed when pulverizing. It is preferable to add water at the time of grinding. The amount of water to be added can be adjusted according to the solids content of the desired soybean in the final fermentation (culture) step, usually 3 to 20 times the weight of the soybeans, preferably 5-15 times. When the amount of water is too small, there is a disadvantage that the grinding efficiency is lowered, and when the amount of water is large, there is a problem that the solid content of the beans of the ground product is lower than the solid content suitable for the final fermentation.

According to a preferred embodiment of the present invention, the soybean meal is obtained by grinding the whole part of the bean. Soy beverages (such as soy milk) that are generally sold are not obtained using the entire portion of the soybean, but rather extracts obtained by hydrothermal extraction of a portion or soybean. These conventional soy drinks do not fully utilize the nutrients the beans have. However, in the present invention, since the pulverized product obtained by pulverizing using all the beans is used as a raw material of the beverage, it is very nutritionally more advantageous than conventional soy beverages. For example, in the present invention, since the medium is prepared using the whole bean without removing the skin of the bean, the finally obtained fermented beverage is rich in natural dietary fiber, improving the intestinal flora and improving constipation. do.

According to a preferred embodiment of the present invention, the method further comprises the step of first sterilizing the medium of step (a), more preferably the grinding liquid. The primary steaming process has the advantage of increasing the efficiency of the use of nutrients by microorganisms in the fermentation of soybeans, and has the effect of preventing the decay of soybean crushed during the enzyme treatment process by sterilizing contaminated various bacteria. Sterilization is preferably carried out for 5-20 minutes at a temperature of 80-121 ℃. If the sterilization time is too long, the color of the final product turns brown due to discoloration due to brown change, which leads to a decrease in commerciality and a reduction in the refreshing aroma generated during fermentation. Abnormal fermentation may occur during the process.

According to a preferred embodiment of the present invention, the soybean pulverized liquid after the first sterilization process is subjected to the enzyme treatment process. This enzyme treatment process, which breaks down the fibers and proteins present in the soybeans and converts them into a form suitable for use by various microorganisms added during the fermentation process. The enzyme used in the enzymatic treatment process is a protease, cellulase or combinations thereof, more preferably a mixed enzyme of protease and cellulase. As such enzyme, it is preferable to use what is marketed for food. The amount of enzyme used is 0.05-1.0% (w / w) based on the total weight of soy solids in the soy flour. When the amount of enzyme is less than 0.05% (w / w), the degradation rate of cellulose or protein is low, and the growth of lactic acid bacteria and Bacillus bacteria is not active during fermentation, whereas the amount of enzyme exceeds 1.0% (w / w). Not only is the processing cost too high, but the excipients used in the enzymes can adversely affect fermentation.

The enzyme treatment process is carried out at 20-80 ° C., preferably 30-80 ° C., more preferably 40-70 ° C., most preferably 40-70 ° C. for 2-24 hours. If the temperature is too low, the activity of the enzyme is weak, on the contrary too high a problem occurs that the activity of the enzyme is lost. If the reaction time is too short, the action of the enzyme is not enough, if too long, a problem arises that the process is expensive.

According to a preferred embodiment of the present invention, a small amount of carbon source is additionally added to the enzymatically treated soy flour. Suitable as further carbon sources of the present invention are selected from the group consisting of glucose, sucrose, maltose, fructose, lactose, xylose, galactose, arabinose and combinations thereof, preferably sucrose, fructose, Glucose, galactose, arabinose and lactose, more preferably sucrose, fructose and glucose, most preferably sucrose.

The amount of additional carbon source is preferably 0.1-20% by weight of the soy flour, more preferably 0.5-10% by weight, most preferably 1-3% by weight.

According to a preferred embodiment of the present invention, the soybean pulverized liquid added with an additional carbon source is subjected to a secondary sterilization process, and pasteurization is preferable in order to minimize nutrient change and browning caused by heat. Secondary sterilization is 10-30 minutes at 70-100 ° C., preferably 80-100 ° C. If the secondary sterilization temperature is low or the time is too short, it is difficult to achieve the desired sterilization effect. If the temperature is too high or the time is too long, there is a problem of nutrient destruction or color change.

One of the characteristics of the present invention is that in culturing vegetable lactic acid bacteria and / or vegetable Bacillus bacteria, only soybean pulverized liquid can be used as a component of the medium. When the above-mentioned additional carbon source is added to the soybean pulverized liquid as a subsidiary material, the microorganisms are cultured, but the number and finality of the final bacteria increase, but the soybean pulverized liquid may partially achieve the object of the present invention. In addition, it is one of the features of the present invention that when adding an additional carbon source to the soybean pulverized liquid and using it as a medium, no other medium component (nitrogen source, minerals, etc.) is required.

(b) inoculating a plant lactic acid bacterium, a plant bacillus bacterium or a mixture thereof

The soybean medium prepared by the above process is inoculated with vegetable lactic acid bacteria, plant Bacillus bacteria or mixed bacteria thereof.

A major feature of the present invention is the use of vegetable lactic acid bacteria and / or vegetable Bacillus bacteria, rather than conventional animal lactic acid bacteria, in preparing soybean fermented beverages.

As used herein, the term “lactic acid bacteria” refers to a group of bacteria that produce lactic acid as a major product of carbohydrate metabolism, eg, Lactococcus , Lactobacillus , Leuconostoc , Pro Rio nibacterium ( Protonibacterium ), Bifidobacterium (Bifidobacterium), Streptococcus (Streptococcus) and Pediococcus , most preferably Lactobacillus.

As used herein, the term “vegetable lactic acid bacteria” means lactic acid bacteria derived from fermented foods of plant raw materials. The vegetable lactic acid bacteria used in the present invention are preferably lactic acid bacteria isolated from kimchi, and more preferably, Lactobacillus sakei , Lactobacillus brevis and Lactobacillus plantarum More preferably, they are Lactobacillus sakei and Lactobacillus plantarum, and most preferably, a mixed lactic acid bacterium of Lactobacillus sakei and Lactobacillus plantarum. Lactobacillus most preferable examples of the Bacillus four Kay, a preferable example of a Lactobacillus four K B2-16 (, KFCC-11321, Republic of Korea Patent No. 0,549,094) isolated from Kimchi by the present inventors, Lactobacillus brevis, the present inventors Lactobacillus brevis B3-18 (Korean Patent No. 0549094) isolated from Kimchi.

As used herein, the term “vegetable Bacillus bacteria” means Bacillus bacteria derived from fermented foods of plant raw materials. The plant Bacillus bacteria used in the present invention are preferably lactic acid bacteria isolated from doenjang, more preferably Bacillus subtilis , and most preferably Bacillus subtilis separated from Chungkookjang by the present inventors. TP6 (Korean Patent Application No. 2005-0008322, KFCC 11343P). Bacillus subtilis TP6 has the ability to ferment carbohydrates contained in soybeans.

Suitable medium for spawning lactic acid bacteria isolated from kimchi is MRS broth (deMan Rogosa Sharpe broth), APT (All Purpose with Tween) medium or BHI (Brain Heart Infusion) medium, most preferably MRS broth medium. The lactic acid bacteria isolated from kimchi in this medium can be incubated at 30 ° C. for 8-16 hours to obtain the desired bacterial activity. In addition, Bacillus subtilis incubated for 12-24 hours at 37 ℃ using a modified nutrition medium (5 g / L Soyton, 5 g / L beef extract and 20 g / L xylose, pH 7.0) It is suitable. Since both Lactobacillus strains and Bacillus bacteria used in the cultivation can be grown in the soybean grinding medium of the present invention, the soybean grinding medium of the present invention may be used for seed culture, and the growth conditions at this time are MRS broth and Same as when using modified nutritional medium.

Lactobacillus isolated from Kimchi has representative characteristics of vegetable lactobacillus. Among them, Lactobacillus sakei, Lactobacillus brevis and Lactobacillus plantarum show strong growth with soybean as a medium, and they have high preference by forming good flavor during growth. Fermented soymilk can be produced. In addition, Bacillus subtilis isolated from soybean paste is typical of the flavor of the doenjang and chungkukjang during solid fermentation over a long time, but in the soybean medium of the present invention grows in a form similar to lactic acid bacteria and has good flavor, so it is not suitable for use as fermentation microorganisms. It is possible.

Lactobacillus sakei, Lactobacillus brevis, Lactobacillus brevis and Lactobacillus plantarum isolated from Kimchi and Bacillus subtilis isolated from doenjang have good symbiotic relationship among strains to produce the desired soybean fermented beverage. There is an advantage to this. In addition, Lactobacillus Sakei strain isolated from Kimchi tends to have a sweet taste, Lactobacillus plantarum has a sour taste, and Bacillus subtilis tends to give a delicious taste. Therefore, in consideration of the contribution to the taste of these strains, by selecting and using the type of strain can be produced soybean fermentation beverages of various preferences.

The inoculation amount of the microorganism is preferably such that the initial bacterial count after inoculation is 10 ⁵ -10 ⁹ cfu / g or 10 ⁵ -10 ⁹ cfu / ml. If the inoculation amount is less than 10 ⁵ cfu / g, it takes a long time to ferment soybean, and the incubation time for the production of the product is long, and if the inoculum exceeds 10 ⁹ cfu / g, burden on the production of spawn seedling .

(c) culturing the inoculated bacteria to obtain a soybean fermented beverage

The bacteria inoculated in the soybean culture are finally cultured to obtain a soybean fermented beverage.

The soybean medium inoculated with the microorganisms is evenly mixed and fermented at a suitable temperature. The fermentation temperature is preferably 20-40 ° C., most preferably about 30 ° C. If it is less than 20 ℃ fermentation does not occur easily, and if it exceeds 40 ℃ there is a problem that can not produce fermented soymilk because the weak lactic acid bacteria do not grow. The time for incubating the lactic acid bacteria is preferably 3-24 hours, more preferably 4-12 hours, and most preferably 6-10 hours.

For details of the culture and fermentation of microorganisms, see Kubitschek, HE, Introduction to Research with Continuous Cultures . Englewood Cliffs, NJ: Prentice-Hall, Inc., 1970; Mandelstam, J., et al., Biochemistry of Bacterial Growth , 3rd ed. Oxford: Blackwell, 1982; Meynell, GG, et al., Theory and Practice in Experimental Bacteriology , 2nd ed. Cambridge: Cambridge University Press, 1970; Gerhardt, P., ed., Manual of Methods for General Bacteriology , Washington: Am. Soc. Microbiol, 1981, which is incorporated herein by reference.

Soybean fermented beverages which have been fermented by vegetable lactic acid bacteria and / or Bacillus bacteria according to the present invention can be commercialized directly into a plain type because they have good palatability in themselves, and the ingredients commonly added to beverages can be added. You can make a variety of products. Soybean fermented beverages of the present invention may include flavoring agents or natural carbohydrates as additional ingredients. For example, natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.); Disaccharides (eg maltose, sucrose, etc.); oligosaccharide; Polysaccharides (eg, dextrins, cyclodextrins, etc.); And sugar alcohols (eg, xylitol, sorbitol, erythritol, and the like). As the flavoring agent, natural flavoring agents (e.g., taumartin, stevia extract, etc.) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) can be used.

The soybean fermented beverage of the present invention is preferably 5 x 10 ⁸ -3.5 x 10 ⁹ cfu / ml, more preferably 1 x 10 ⁹ -3.5 x 10 ⁹ cfu / ml, even more preferably 1.5 x 10 ⁹ -3.5 x 10 ⁹ cfu / ml, most preferably 2 x 10 ⁹ -3.5 x 10 ⁹ cfu / ml.

The pH of the soybean fermented beverage of this invention becomes like this. Preferably it is 4.0-5.0, More preferably, it is 4.2-4.8, Most preferably, it is 4.3-4.6.

The soybean fermented beverage of the present invention is not only excellent in flavor compared to conventionally fermented soy milk, but also contains a large number of bacteria of lactic acid bacteria and / or Bacillus which are known to be beneficial to the human body. In addition, since the soybean fermented beverage of the present invention was prepared using the whole soybean without removing the husks of the soybeans, all of the soybean nutrients are included, in particular, rich in natural dietary fiber to improve the intestinal flora and improve constipation It is effective. In addition, since the fermented strains are vegetable lactic acid bacteria and Bacillus bacteria isolated from kimchi and doenjang, rather than the existing animal lactic acid bacteria, soybean fermented beverage of the present invention can be said to have the advantage of being made of 100% domestic materials.

In addition, by using various types of soybeans, which have many functional and nutritional ingredients, but have a limited form of consumption such as soy milk, tofu, etc., or mixed with rice, the present invention provides preference and functionality. It means that it contributes to people's health by developing products that all have.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention more specifically, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example

Throughout this specification, unless otherwise indicated, “%” used to indicate the concentration of a particular substance is solid / solid (weight / weight)%, solid / liquid (weight / volume)%, and Liquid / liquid is (volume / volume)%.

Example  1: Preparation of Soybean Medium for Soybean Fermented Beverage Production

Using soybeans purchased from the Agricultural Cooperatives, the product was washed three times in cold water to remove dust and foreign matter from the surface. 1 kg of washed soybeans were soaked in 4 L of cold water for 12 hours. 6 L of water was further added to the soaked soybeans, and then pulverized into 200 mesh using a pulverizer, followed by primary sterilization for 10 minutes at 110 ° C.

After the sterilized soybean pulverized product was cooled to 50 ° C, 1 g of cellulase (Bioland) and 1 g of protease (Bioland) were added, and left at 55 ° C for 2 hours. A portion of the cellulose component and the protein component was decomposed and converted into a form readily used by the lactic acid bacteria and / or Bacillus bacteria added.

20 g of sucrose was added to the enzyme-treated soybean pulverized solution, followed by secondary sterilization at 80 ° C. for 15 minutes to complete the soymilk medium.

Example  2: Kimchi lactic acid bacteria  And Bacillus  Production of Fermented Soymilk Using Bacteria

Lactobacillus sakei incubated for 18 hours in MRS broth (deMan Rogosa Sharpe broth) in four soymilk medium prepared in 1 L of Erlenmeyer flasks (prepared in Example 1). B2-16 , KFCC-11321, Korean Patent No. 0549094) , Lactobacillus brevis B3-18, Bioben Co., Korea Patent No. 0549094) and Lactobacillus plantarum P-23 and Bioben) were each inoculated with a final bacterial concentration of 10 ⁸ cfu / g and incubated for 12 hours. Bacillus subtilis also cultured for 16 hours in modified nutrient medium (5 g / L Soyton, 5 g / L Beef Extract and 20 g / L Xylose, pH 7.0). TP6, Republic of Korea Patent Application No. 2005-0008322, KFCC 11343) was inoculated in soybean medium so that the final bacteria concentration is 10 ⁸ cfu / g and incubated for 8 hours. The temperature was maintained at 30 ° C. and cultured. At this time, the initial pH of the medium was 6.3. The pH and viable cell number of the fermented soymilk are listed in Table 1.

All three Lactobacillus bacteria used in this experiment were isolated from Kimchi and Bacillus subtilis bacteria were isolated from Doenjang.

Inoculation strain Cell number after fermentation (cfu / ml) PH after fermentation Remarks Lactobacillus sakei 1.7 × 10 ⁹ 4.6 Kimchi isolate Lactobacillus brevis 9.5 × 10 ⁸ 4.5 Kimchi isolate Lactobacillus plantarum 2.5 × 10 ⁹ 4.3 Kimchi isolate Bacillus subtilis 1.1 × 10 ⁹ 4.6 Miso isolate

As shown in Table 1, pH of the final product was all there was no significant difference in the Fermented Milk and Milk existing materials in the range of 4.2 to 4.8, all bacteria may be enough to hold the 10 conditions as indicated for ⁸ cfu / ml or more fermented beverages I could see that.

Example  3: lactic acid bacteria and Bacillus bacteria  Production of Fermented Soymilk by Mixed Culture

Lactobacillus sakei incubated in MRS broth (deMan Rogosa Sharpe broth) for 18 hours B2-16 , KFCC-11321, Korean Patent No. 0549094) , Lactobacillus brevis B3-18, Bioben Co., Korea Patent No. 0549094) and Lactobacillus plantarum P-23 , Bioben Co.) and Bacillus subtilis ( Bacillus subtilis , Korean Patent Application No. 2005-0008322, KFCC 11343) incubated for 18 hours in modified nutrient medium, Example 2-4 Soymilk medium prepared in the same manner as 1 was inoculated so that the final bacteria concentration 3 × 10 ⁸ cfu / g and incubated for 8 hours.

Evaluation of pH, viable cell number, and taste of fermented soymilk using mixed strains is summarized in Table 2. The evaluation of taste showed the result which carried out the perfect score of 5 points using 7 panelists.

Inoculation strain Cell number after fermentation (cfu / ml) PH after fermentation Sensory evaluation Sour taste sweetness bitter Astringent Sweet taste Lactobacillus sakei Lactobacillus brevis 2.7 × 10 ⁹ 4.5 3.5 3.6 4.0 3.5 4.0 Lactobacillus sakei Lactobacillus plantarum 3.1 × 10 ⁹ 4.3 4.0 4.1 3.5 4.0 4.1 Lactobacillus sakei Bacillus subtilis 9.8 × 10 ⁸ 4.6 3.0 3.3 4.3 3.3 3.8 Lactobacillus brevis Lactobacillus plantarum 2.5 × 10 ⁹ 4.5 3.3 3.2 3.5 3.4 3.9 Lactobacillus brevis bacillus subtilis 6.4 × 10 ⁸ 4.3 3.0 3.1 3.1 4.0 4.2 Lactobacillus plantarum bacillus subtilis 2.7 × 10 ⁹ 4.2 3.6 3.4 3.3 3.6 4.1 Lactobacillus sakei Lactobacillus brevis Lactobacillus plantarum 2.2 × 10 ⁹ 4.3 4.1 3.5 3.2 3.5 3.9 Lactobacillus brevis Lactobacillus plantarum bacillus subtilis 1.8 × 10 ⁹ 4.4 4.2 3.3 4.0 3.5 4.0 Lactobacillus sakei Lactobacillus plantarum bacillus subtilis 2.5 × 10 ⁹ 4.4 4.3 4.1 3.9 4.2 4.4 Lactobacillus sakei Lactobacillus brevis Lactobacillus plantarum bacillus subtilis 1.7 × 10 ⁹ 4.3 3.9 4.0 3.6 3.4 3.9

* Sensory evaluation: 1; Very bad, two; Bad; 3; Normal, 4; Good five; Very good

As shown in Table 2, the soybean medium prepared by the present invention does not significantly change the degree of preference even when inoculated from 2 to 4 species of lactic acid bacteria isolated from Kimchi and Bacillus bacteria isolated from Cheonggukjang and excellent symbiotic relationship between the strains. There was no problem in making fermented soymilk. Especially in the sensory evaluation, Lactobacillus Sakei strains showed a sweet taste, Lactobacillus plantarum showed a sour taste, and Bacillus subtilis tended to give a delicious taste. Lactobacillus brevis showed high viable cell numbers.

In addition, it can be seen that the strain used in the present invention can be used alone or in combination in the preparation of soybean fermented beverages by the effect of the reinforcement of the characteristics of each bacteria by mixing the strains.

Example  4: preparation of fermented beverage using various beans

Soybeans, mung beans, black beans, seomoktae, peas and kidney beans were purchased from the Agricultural Cooperatives and washed three times in cold water to remove dirt and foreign matter from the surface. 100 g of washed beans were soaked in 1 L of cold water for 10 hours. 2 L of water was added to the immersed soybean, and then pulverized into 300 mesh using a pulverizer, followed by primary sterilization at 110 ° C. for 10 minutes.

After cooling the sterilized soybean pulverized product at 50 ° C., 0.5 g of cellulase and 0.5 g of protease were added, and then left at 60 ° C. for 4 hours, and then 3 g of sucrose was added. Tea sterilization was completed to complete the medium using various beans.

Lactobacillus sakei incubated in deMan Rogosa Sharpe broth for 15 hours in sterilized soymilk medium B2-16 , KFCC-11321) and Lactobacillus plantarum p-23, Bioben Co., Ltd. and Bacillus subtilis (Korean Patent Application No. 2005-0008322, KFCC 11343) incubated for 18 hours in a modified nutrient medium in an equal volume of 8.0 x 10 ⁷ Inoculated to cfu / ml and incubated for 9 hours.

 Table 3 shows the pH, viable cell count and preference of soybean fermented beverages of various soybean raw materials. The sensory evaluation result is a result of evaluating comprehensive preference degree using a panel of seven people with a perfect score of 5 points.

Kinds pH Cell number after fermentation (cfu / ml) Sensory evaluation Big head 4.38 2.7 × 10 ⁹ 4.5 green gram 4.43 1.1 × 10 ⁹ 3.8 Black Beans 4.48 9.2 × 10 ⁸ 4.3 Seomoktae 4.52 5.5 × 10 ⁸ 4.2 pea 4.62 6.1 × 10 ⁸ 3.7 kidney bean 4.58 4.7 × 10 ⁸ 3.5

* Sensory evaluation: 1; Very bad, two; Bad; 3; Normal, 4; Good five; Very good

As shown in Table 3, soybean fermented beverages could be prepared using various types of beans. Soybean, black soybean, and Seomoktae were good in preference. In case of kidney beans and peas, the preference was slightly decreased, but drinking was possible, and the number of viable cells showed sufficient commerciality as fermented beverage.

Example  5: Microbial Changes During Storage of Fermented Soymilk

Soybeans purchased from the Agricultural Cooperatives were washed in cold water to remove dirt and foreign matter from the surface. After washing, 2 kg of soybeans were soaked in 15 L of cold water for 12 hours. 6 L of water was added to the immersed soybeans, and then pulverized into 250 mesh using a pulverizer, followed by primary sterilization for 15 minutes at 110 ° C.

After cooling the sterilized soybean pulverized product to 55 ° C, add 1 g of cellulase and 1 g of protease, and leave it at 60 ° C for 4 hours, add 30 g of sucrose, and then secondary at 80 ° C for 15 minutes. Sterilization to complete the soymilk medium.

Lactobacillus sakei incubated in deMan Rogosa Sharpe broth for 15 hours in sterilized soymilk medium B2-16 , KFCC-11321) and Lactobacillus plantarum p-23, Bioben Co., Ltd. and Bacillus subtilis (Korean Patent Application No. 2005-0008322, KFCC 11343) incubated for 16 hours in a modified nutritional medium were mixed in the same volume to obtain a final bacterial concentration of 3 × 10 ^8. Inoculated to cfu / ml and incubated for 7 hours.

 The culture and fermented soymilk were stored at 10 ° C. for 30 days. The sensory evaluation result is a result of evaluating comprehensive preference degree using a panel of seven people with a perfect score of 5 points.

Storage days pH Final cell count (cfu / ml) Sensory evaluation 0 4.48 1.60 × 10 ⁹ 4.5 One 4.43 2.30 × 10 ⁹ 4.6 2 4.28 1.50 × 10 ⁹ 4.6 5 4.22 1.20 × 10 ⁹ 4.6 7 4.2 1.00 × 10 ⁹ 4.5 10 4.29 2.10 × 10 ⁹ 4.4 15 4.33 1.30 × 10 ⁹ 4.5 20 4.28 1.80 × 10 ⁹ 4.3 30 4.14 4.50 × 10 ⁹ 4.3

* Sensory evaluation: 1; Very bad, two; Bad; 3; Normal, 4; Good five; Very good

As shown in Table 4, even after 30 days of fermentation, the viable cell count was maintained at 10 ⁹ cfu / ml or more, and the pH was maintained at 4.1 or higher, so there was no problem in drinking, and there was no significant decrease in storage. It was found that distribution was possible for more than 30 days after production.

Example  6: Enzyme Treatment Effect during Soymilk Medium Preparation

Soybeans purchased from the Agricultural Cooperatives were washed twice with cold water to remove dirt and foreign matter from the surface. 1 kg of washed soybeans were immersed in 5 L of cold water for 6 hours. 6 L of water was added to the immersed soybeans, and then pulverized into 300 mesh using a pulverizer, followed by primary sterilization for 15 minutes at 110 ° C.

After cooling the sterilized soybean pulverized product to 50 ° C., 1 g of cellulase and 1 g of protease were added alone or in a mixture, and then left at 60 ° C. for 4 hours, and then 20 g of sucrose was added, followed by 15 at 80 ° C. Sterilization for 2 minutes to complete the soymilk medium. In addition, the second sterilized soymilk medium was prepared immediately without addition of enzyme and used as a control.

Lactobacillus plantarum incubated in MRS broth (deMan Rogosa Sharpe broth) for 15 hours in each sterilized soymilk medium p-23, Bioben Co., Ltd. and Bacillus subtilis (Korean Patent Application No. 2005-0008322, KFCC 11343) incubated for 18 hours in modified nutrient medium at a ratio of 1: 0.7 (v: v) Table 5 shows the results of incubation for 6 hours after inoculation with a final bacterial concentration of 1.5 × 10 ⁸ cfu / ml.

Used enzymes Cell number after fermentation (cfu / ml) PH after fermentation Sensory evaluation Sour taste sweetness bitter Astringent Sweet taste Protease 1.7 × 10 ⁹ 4.6 3.5 3.5 3.2 3.5 4.1 Cellulase 1.9 × 10 ⁹ 4.6 3.7 3.5 3.3 3.8 3.9 Protease + Cellulase 2.1 × 10 ⁹ 4.4 3.6 3.4 3.3 3.6 4.1 No treatment 8.5 × 10 ⁸ 4.8 2.6 2.1 2.9 1.9 2.4

* Sensory evaluation: 1; Very bad, two; Bad; 3; Normal, 4; Good five; Very good

As shown in Table 5, when the enzyme was not treated, soybean utilization decreased and the number of bacteria in the final fermented soymilk decreased, as well as a low value in the degree of preference. Therefore, in order to fully exhibit the effects of the present invention, it can be said that it is necessary to treat proteases and cellulase during the preparation of the medium. Each enzyme did not show a big difference, it could be seen that the two enzymes can be used alone or in combination.

Example  7: Changes in the Characteristics of Soybean Fermented Milk According to the Type of Adjunct

Soybeans purchased from the Agricultural Cooperatives were washed twice with cold water to remove dirt and foreign matter from the surface. 1 kg of washed soybeans were immersed in 5 L of cold water for 12 hours. 6 L of water was further added to the soaked soybeans, and then pulverized into 150 mesh using a pulverizer, followed by primary sterilization for 15 minutes at 110 ° C.

After cooling the sterilized soybean pulverized to 55 ℃, 1 g of cellulase and 1 g of protease was added and left at 60 ℃ for 4 hours and then sucrose, fructose, glucose, galactose, Lactose and a carbon source consisting of two or more of them were added so as to have a total added amount of 30 g at the same ratio, followed by secondary sterilization at 80 ° C. for 15 minutes to complete the soybean medium.

Lactobacillus sakei B2-16 , KFCC-11321, Lactobacillus plantarum p-23 (Bioben) and Bacillus subtilis (Korean patent) Application No. 2005-0008322, KFCC 11343) was mixed in the same volume and inoculated so that the final bacterial concentration is 1 x 10 ⁸ cfu / ml, and the results of incubation for 6 hours are shown in Table 6.

Carbon source Cell number after fermentation (cfu / ml) PH after fermentation Sensory evaluation Sour taste sweetness bitter Astringent Sweet taste Sucrose 2.7 × 10 ⁹ 4.4 3.5 4.6 4.0 3.5 4.0 Fructose 2.1 × 10 ⁹ 4.3 4.0 4.1 3.5 4.0 4.1 Glucose 9.2 × 10 ⁸ 4.6 3.0 3.3 4.3 3.3 3.8 Galactose 1.5 × 10 ⁹ 4.5 3.3 3.2 3.5 3.4 3.9 Lactose 6.4 × 10 ⁸ 4.7 3.0 3.1 3.1 3.5 3.2 Sucrose + fructose 2.4 × 10 ⁹ 4.3 4.3 4.4 4.3 3.6 4.1 Sucrose + glucose 2.1 × 10 ⁹ 4.3 4.1 4.2 3.2 3.5 3.9 Sucrose + galactose 1.8 × 10 ⁹ 4.4 3.6 3.3 4.0 3.5 4.0 Sucrose + lactose 2.2 × 10 ⁹ 4.4 4.0 4.1 3.9 4.2 4.0 Sucrose + fructose + glucose 1.5 × 10 ⁹ 4.4 3.9 4.0 3.6 3.4 3.9 Sucrose + fructose + galactose 2.3 × 10 ⁹ 4.3 3.5 3.6 4.0 3.5 4.0 Sucrose + fructose + lactose 3.1 × 10 ⁹ 4.3 4.0 4.0 3.5 4.0 4.1 Sucrose + fructose + glucose + galactose 9.8 × 10 ⁸ 4.6 3.0 3.5 4.3 3.3 3.8 Sucrose + fructose + glucose + lactose 2.5 × 10 ⁹ 4.5 3.3 3.3 3.5 3.4 3.9 Sucrose + fructose + glucose + galactose + lactose 6.4 × 10 ⁸ 4.3 3.8 3.1 3.1 4.0 4.2

* Sensory evaluation: 1; Very bad, two; Bad; 3; Normal, 4; Good five; Very good

As shown in Table 6, it can be seen that the subsidiary material as a carbon source added to the soybean grinding medium prepared according to the present invention is not significantly affected by the type and combination thereof. The high viable cell counts were found in all carbon sources alone and in the mixed use group, and the organoleptic results showed above average results in all combinations.

Example  8: Change of Characteristics of Soybean Fermented Beverages According to Addition of Supplementary Ingredients

1 kg of soybean purchased from Nonghyup was washed in cold water and then immersed in 12 L of cold water for 12 hours. After soaking, the soybean was pulverized into 200 mesh using a pulverizer and subjected to primary sterilization at 110 ° C. for 15 minutes.

After cooling the sterilized soybean pulverized product at 60 ° C, add 0.7 g of cellulase and protease, and leave it at 60 ° C for 4 hours, and then add sucrose and fructose according to the concentration. Sterilization to complete the soymilk medium.

Lactobacillus sakei incubated in deMan Rogosa Sharpe broth for 15 hours in sterilized soymilk medium B2-16 , KFCC-11321) and Lactobacillus plantarum p-23, Bioben Co., Ltd. and Bacillus subtilis cultured in modified medium nutrient medium for 18 hours were mixed in the same volume and inoculated so that the final bacterial concentration was 1.5 × 10 ⁸ cfu / ml, followed by incubation for 6 hours. The results are shown in Table 7. The sensory evaluation result is a result of evaluating comprehensive preference degree using a panel of seven people with a perfect score of 5 points.

Raw materials Addition amount (w / v) pH Cell number after fermentation (cfu / ml) Sensory evaluation Sucrose 0 4.78 7.60 × 10 ⁸ 3.3 0.5 4.63 1.10 × 10 ⁹ 4.2 One 4.58 1.50 × 10 ⁹ 4.4 2 4.50 1.20 × 10 ⁹ 4.5 3 4.48 2.00 × 10 ⁹ 4.7 5 4.48 2.10 × 10 ⁹ 4.3 10 4.45 2.30 × 10 ⁹ 4.2 Fructose 0 4.79 7.60 × 10 ⁸ 3.3 0.5 4.64 1.00 × 10 ⁹ 4.2 One 4.57 1.54 × 10 ⁹ 4.4 2 4.49 1.29 × 10 ⁹ 4.4 3 4.47 2.10 × 10 ⁹ 4.6 5 4.48 2.10 × 10 ⁹ 4.5 10 4.44 2.30 × 10 ⁹ 4.1

* Sensory evaluation: 1; Very bad, two; Bad; 3; Normal, 4; Good five; Very good

As shown in Table 7, even when no additional ingredients were added, the bacteria added by the nutrients decomposed by the enzymes were grown, but the growth degree was weaker and the sensory values were much lower than those when the additional ingredients were added. . In addition, even when the amount of carbon source was increased, there was no substrate inhibition effect, but the sweetness was too strong due to the unused substrate.

Example  9: Preparation of Staple Bean Fermented Beverage

1 kg of black beans purchased from the NH was washed in cold water and then immersed in 8 L of cold water for 10 hours. After soaking, the soybeans were pulverized into 300 mesh using a pulverizer and subjected to primary sterilization for 15 minutes at 110 ° C.

After cooling the sterile black soybean pulverized product to 55 ℃, add 2 g of cellulase and 1 g of protease, and leave it at 60 ℃ for 8 hours, add sucrose and glucose by concentration, and then at 80 ℃ for 15 minutes The second sterilization completed the black soybean medium.

Lactobacillus sakei incubated in deMan Rogosa Sharpe broth for 15 hours in sterilized soymilk medium B2-16 , KFCC-11321) and Lactobacillus plantarum p-23, Bioben Co., Ltd. and Bacillus subtilis (Korean Patent Application No. 2005-0008322, KFCC 11343) incubated for 15 hours in modified nutrient medium were mixed to the same volume to obtain a final bacterial concentration of 4 x 10 ⁸ After inoculating to cfu / ml to incubate for 8 hours to complete a black bean fermented beverage that can be eaten with a spoon.

As described in detail above, the present invention provides a method for producing a soybean fermented beverage using vegetable lactic acid bacteria and / or Bacillus as a raw material and soybean fermented beverage.

The soybean fermented beverage of the present invention is not only excellent in flavor compared to conventionally fermented soy milk, but also contains a large number of bacteria of lactic acid bacteria and / or Bacillus which are known to be beneficial to the human body. In addition, since the soybean fermented beverage of the present invention was prepared using the whole soybean without removing the husks of the soybeans, all of the soybean nutrients are included, in particular, rich in natural dietary fiber to improve the intestinal flora and improve constipation It is effective. In addition, since the fermented strains are vegetable lactic acid bacteria and Bacillus bacteria isolated from kimchi and doenjang, rather than the existing animal lactic acid bacteria, soybean fermented beverage of the present invention can be said to have the advantage of being made of 100% domestic materials.

In addition, by using various types of soybeans, which have many functional and nutritional ingredients, but have a limited form of consumption such as soy milk, tofu, etc., or mixed with rice, the present invention provides preference and functionality. It means that it contributes to people's health by developing products that all have.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (16)

Soybean fermented beverage manufacturing method comprising the following steps: (a) preparing a medium comprising soy flour obtained by milling all parts of the soybeans; (b-1) enzymatically treating the medium of step (a) with a protease, cellulase or a combination thereof; (b-2) inoculating the culture of lactic acid bacteria, plant Bacillus or mixed bacteria thereof to the medium of step (b-1); And (c) culturing the bacteria inoculated in the medium to obtain a soybean fermented beverage.        The method of claim 1, wherein the beans are soybeans, green beans, black beans, Seomoktae, peas or kidney beans.        The method of claim 1, wherein step (a) is performed by pulverizing soybeans called water to a size of 20-200 μm.        The method of claim 1, wherein the method further comprises the step of first sterilizing the medium of step (a) at 80-121 ° C. for 5-20 minutes. delete delete        The method of claim 1, wherein the medium further comprises a carbon source selected from the group consisting of glucose, sucrose, maltose, fructose, lactose, xylose, galactose, arabinose and combinations thereof.       The method of claim 1, wherein the vegetable lactic acid bacteria is a Lactobacillus strain isolated from Kimchi. The method of claim 8, wherein the Lactobacillus sp. Strain is Lactobacillus sakei , Lactobacillus brevis or Lactobacillus plantarum . The method according to claim 1, wherein the Bacillus bacteria is a strain of the genus Bacillus isolated from doenjang. The method of claim 1, wherein the inoculum of the plant lactic acid bacteria, plant Bacillus bacteria or mixed bacteria thereof is 10 ⁵ -10 ⁹ cfu / g or 10 ⁵ -10 ⁹ cfu / ml. The method of claim 1, wherein the culturing is carried out at 20-40 ℃.        The method of claim 1, wherein said culturing is carried out for 3-24 hours. A soybean fermented beverage prepared by the method of any one of claims 1 to 4 and 7 to 13, wherein the soybean fermented beverage comprises a vegetable lactic acid bacterium, a vegetable Bacillus bacteria or a mixture thereof Fermented soybean drink. ^15. The soybean fermented beverage of claim 14, wherein the soybean fermented beverage has a viable cell count of 5 x 10 ⁸ -3.5 x 10 ⁹ cfu / ml. 15. The soya fermented beverage according to claim 14, wherein the soybean fermented beverage has a pH of 4.0-5.0.

Images