KR101171401B1 - Method of manufacturing liquid koji - Google Patents

Abstract

An object of the present invention is to provide a liquid station with high enzymatic activity of a glucoamylase and an acid resistant α-amylase which can be used for the production of fermented foods and drinks, especially for brewing shochu.

The present invention relates to a method for producing a liquid station used in the production of fermented food and beverages, as a raw material for cereals covered with cereals, cereals (only chaff, etc.) removed from the surface, raw beans or potatoes covered with the skin, And culturing the bacterium in a liquid medium comprising any one of amaranthus and / or quinoa in cereals.

According to the present invention, it is possible to produce a liquid station in which both enzymes of glucoamylase and acid resistant α-amylase are simultaneously produced in good balance and having the enzyme activity necessary for brewing shochu. By using this liquid station, fermented foods and beverages such as shochu can be efficiently produced.

Liquid soup, shochu brewing, glucoamylase, acid resistant α-amylase

Description

Method for manufacturing liquid station {Method of manufacturing liquid koji}

TECHNICAL FIELD This invention relates to the manufacturing method of the liquid station used for manufacture of fermented food-drinks, especially the liquid station which has the enzyme activity required for brewing shochu.

The soup used in the production of alcoholic beverages is a solid state in which the spores of filamentous fungi are inoculated and cultured on raw materials after steaming or the like, and raw materials in water. And other nutrient sources to prepare a liquid medium, and there are liquid stations inoculated and cultured with spores of mycelium or mycelium pre-cultured.

In the manufacture of conventional liquor or fermented food or drink, for example, Japanese sake, shochu, soy sauce, miso, sweet sake, etc., so-called solids impregnated by a solid culture method Bureau is widely used. This solid culture method is aspergillus kawachii, aspergillus awamori, aspergillus niger, aspergillus niger, aspergillus oryzae. ) Or spores of domestic bacteria such as Aspergillus sojae are sprinkled with solid raw materials such as increased grains, and are grown on the surface thereof.

For example, in the production of soju, solid countries such as Aspergillus kawachii and Aspergillus awamori are widely used. However, since the solid culture method is a culture system in which raw materials and Korean bacteria are uniformly dispersed, it is difficult to uniformize factors such as temperature, water content, and various nutrients, and the culture control is very difficult. Troublesome In addition, in many cases, the empire is opened in an open state, and in this case, attention must be paid in terms of quality control such as contamination by various germs. Therefore, it can also be said that it is not suitable for large scale manufacturing.

In contrast, the liquid culture method is a culture form suitable for efficient production because of easy culture control and quality control, but a culture obtained by liquid culture of Korean bacteria, for example, due to insufficient enzymatic activity necessary for brewing shochu. There are few examples of actually using as a minority country. Herein, the culture obtained by the liquid culture method may be a culture solution, cells, their concentrates or their dried product, in addition to the culture itself obtained by the liquid culture method (hereinafter also referred to as a liquid station).

As a large reason why the culture obtained by the liquid culture method is not used for the production of fermented foods and beverages such as soju, in the liquid culture, not only the production of enzymes such as amylase and cellulase of Korean bacteria differs greatly from the solid culture, but overall productivity is high. It is known to fall (refer nonpatent literature 1).

Usually, in the production of alcoholic beverages, including soju, alcohol is produced by parallel fermentation. Therefore, the bacterium glycolysis-related enzymes, particularly glucoamylase and acid-stable α-amylase, which affect glucose supply to the bacterium, are key enzymes in alcohol fermentation. However, in the culture obtained by the liquid culture method, it is known that the activity of glucoamylase is remarkably low, and production behavior differs greatly from solid culture (refer nonpatent literature 2).

As a method for improving the glucoamylase activity of Korean bacteria, a method of culturing Korean bacteria while stressing the growth of mycelia (see Patent Document 1) or a method of adding edible grains to the Korean culture medium (see Patent Document 2). ) Is reported. The method disclosed in Patent Document 1 is to increase the same enzyme activity by expressing a novel gene glaB encoding glucoamylase by culturing in an inclusive fixing agent having a porous membrance or an aperture. Rigorous control or special incubators are required and not practical. Moreover, the method disclosed by patent document 2 is culture | cultivated a domestic bacterium in the liquid medium using the cereal grain which carried out at least one part of a raw material, and the new manufacturing process of culturing a grain becomes added.

Therefore, the present inventors proposed the invention regarding the cultivation method of the bacterium using the liquid medium containing hardly degradable saccharide in a bacterium (refer patent document 3). According to the present invention, it is possible to easily and inexpensively obtain a domestic bacteria culture having high activity of glycolysis-related enzymes such as glucoamylase, which can be used for the production of alcoholic beverages or fermented foods and beverages.

On the other hand, about the acid-resistant (alpha)-amylase, molecular biological analysis is started in detail recently (refer nonpatent literature 3). According to him, Baekkukyun has two types of amylase genes, which differ in properties such as non-acid-resistant α-amylase and acid-resistant α-amylase, but their expression patterns differ greatly, and in liquid culture, non-acid-resistant It has been reported that α-amylase is sufficiently produced, but little acid-resistant α-amylase, which is a key enzyme of shochu brewing, is produced.

In shochu production, it is brewed in a low pH environment to prevent putrefaction of shochu mash. However, since the non-acid-resistant α-amylase is rapidly inactivated under low pH conditions, it hardly contributes to the glycolysis of shochu brewing. Therefore, it is indispensable for the production of soju to produce a large amount of acid resistant α-amylase, which is thought to contribute to the glycolysis of shochu brewing, by liquid culture of Korean bacteria.

In the past, there have been reports on the production behavior of acid resistant α-amylase in liquid culture of Korean bacteria, but the method uses a synthetic medium containing peptone or citric acid buffer, and the culture time takes 100 hours or more. It is hard to say that it is a manufacturing method of the liquid station which can be applied to actual shochu brewing (refer nonpatent literature 4).

Patent Document 1: Japanese Patent Application Laid-Open No. 11-225746

Patent Document 2: Japanese Patent Application Laid-Open No. 2001-321154

Patent Document 3: Japanese Unexamined Patent Publication No. 2003-265165

[Non-Patent Document 1] Iwashita K. et al: Biosci. Biotechnol. Biochem., 62, 1938-1946 (1998), Yamane Yuichi et al .: Japan Brewing Association., 99, 84-92 (2004)

[Non-Patent Document 2] Hata Y. et al: J. Ferment. Bioeng., 84, 532-537 (1997), Hata Y. et al: Gene., 207, 127-134 (1998), Ishida H. et al: J. Ferment. Bioeng., 86, 301-307 (1998), Ishida H. et al: Curr Genet., 37, 373-379 (2000)

[Non-Patent Document 3] Nagamine K. et al: Biosci. Biotechnol. Biochem., 67, 2194-2202 (2003)

[Non-Patent Document 4] Sudo S. et al: J. Ferment. Bioeng., 76, 105-110 (1993), Sudo S. et al: J. Ferment. Bioeng., 77, 483-489 (1994), Shigeto-shi Sudo, etc .: Japan Brewing Association Magazine, 89, 768-774 (1994)

DISCLOSURE OF INVENTION

Problems to be solved by the invention

However, in the method of Patent Literature 3, a Korean culture having a high activity of glucoamylase is a culture of Korean bacteria in a liquid medium prepared by adding hardly degradable saccharide, and a normal liquid medium prepared from culture materials such as cereals is used. It is not cultured using.

In addition, although a technique for obtaining a bacterium culture having high glucoamylase activity by culturing the bacterium in a liquid medium has been disclosed, a liquid medium having a high activity of acid-resistant α-amylase, which is another key enzyme in alcohol fermentation, is used as a liquid medium. It is not disclosed that the technique obtained by culturing the bacterium in Korea. It is generally said that this acid resistant alpha-amylase is an enzyme which is not produced | generated in liquid culture, and the liquid station with high activity of acid resistant alpha-amylase has not been developed until now.

An object of the present invention is to add a special sugar or the like to a liquid station used in the manufacture of fermented food and drink, particularly a liquid station having high activity of glucoamylase and acid resistant α-amylase, which are key enzymes in alcohol fermentation of brewing shochu, It is not a special liquid medium that uses a raw material that has been treated with pears, but as a raw material, grains of white rice, that is, grains whose surface is covered with husk, or grains from which only the surface grains (chaff, etc.) are removed, It is to provide a method for producing a liquid station by culturing the bacterium in a liquid medium using raw beans or potatoes, or specific grains.

Means for solving the problem

As a result of intensive studies to solve the above problems, the present inventors and the like, in the production of a liquid station for use in the production of fermented food and drink, as described above, cereals covered with cereals or surface grains as a culture raw material. Glucoamylase activity and acid resistance by culturing the bacterium in liquid medium containing only grains (such as chaff), soybeans or potatoes covered with shells, or amaranthhus and / or quinoas of cereals It has been found that liquid stations with enhanced α-amylase activity are produced. Moreover, the liquid station which enhanced this enzyme activity discovered that it is suitable for brewing shochu, and came to complete this invention.

That is, this invention provides what is shown below.

(1) A method for producing a liquid station for use in producing fermented food or beverage, comprising: cereal grains covered with a surface of grains as a culture raw material; Cereals from which only the surface grains (chaff etc.) are removed; Raw legumes or potatoes covered with an outer sheath; And culturing the bacterium in a liquid medium comprising any one of Amaranthus and / or Quinoa.

(2) The method for producing a liquid state according to (1), wherein the bacterium is Baekkukyun and / or Black Bacillus.

(3) The method for producing a liquid station according to (1), wherein the grain covered with the surface of the grain is equal to or greater than the whitening ratio to which white grain or at least grain is left on the surface of the kernel.

(4) The method for producing a liquid station according to (1) or (3), wherein the cereals are barley.

(5) The method for producing a liquid state according to (4), wherein the surface barley has the aforementioned whitening ratio of 90% or more.

(6) The method of producing a liquid state according to (1) or (3), wherein the cereals are rice, wheat, buckwheat, barnyard millet, foxtail millet, millet, kaoliang or corn. .

(7) The method for producing a liquid station according to (1), wherein the grains obtained by removing only surface grains of rice (rice husk, etc.) are brown rice.

(8) The method for producing a liquid soup according to (1), wherein the raw beans or potatoes covered with the outer skin are soybeans, red beans, or sweet potatoes.

(9) The method for producing a liquid state according to (1), wherein Amaranthus and / or Quinoa are not subjected to pretreatment such as grinding or crushing.

(10) The liquid station according to any one of (1) to (9), wherein at least a glucoamylase and an acid resistant α-amylase are simultaneously produced and accumulated in a domestic culture cultured in a liquid medium containing the above-mentioned culture raw material. Manufacturing method.

(11) A method for producing a fermented food or drink, wherein the fermented food or drink is produced using the liquid station obtained by any of the methods (1) to (10).

(12) The method for producing a fermented food or drink according to (11), wherein the production of the fermented food and drink is carried out in a liquid phase.

(13) The method for producing a fermented food or drink according to (11) or (12), wherein the production of the fermented food or drink is performed in a liquid phase in a state where an external system and a shielding state are maintained.

(14) The method for producing a fermented food or drink according to any one of (11) to (13), wherein the production of the fermented food or drink is performed by adding a raw material to the liquid station to produce a primary mash.

(15) The method for producing a fermented food or drink according to any one of (11) to (14), wherein the fermented food and drink is shochu.
(16) A liquid station set for the production of fermented food and drink having at least a glucoamylase activity and an acid resistant α-amylase activity obtained by the method for producing a liquid station in any one of (1) to (10).
(17) The rate of release of sugars derived from starch in the culture material into the culture system according to any one of (1) to (9), in the production of a liquid station for culturing Korean bacteria in a liquid medium containing the culture material described above. A method for producing a liquid station, in which the enzyme activity of the liquid station is adjusted by suppressing

Effects of the Invention

According to the present invention, by culturing the bacterium in a liquid medium containing a variety of the above-mentioned culture raw materials, a liquid state in which a group of enzymes necessary for brewing shochu, such as glucoamylase or acid-resistant α-amylase, is produced at the same time, is produced. can do. Since liquid culture enables precise culture control as compared to solid culture, it is possible to produce a liquid station with stable quality and to manufacture it efficiently.

In addition, the use of the liquid station prepared according to the present invention results in the same level of fermentation as the soju mash using the conventional solid station, and the soju prepared has the same quality as the soju prepared using the solid station. Soju can be produced in a sensual sense.

In addition, since the culture raw material used in the present invention is whitened to unwhitened, unprocessed, or at least to the extent that the outer skin remains on the surface, it is expected to improve raw material utilization and yield.

In the case of producing shochu using the liquid station prepared according to the present invention, since the whole process can be performed in a liquid state, unlike the conventional preparation of shochu using a solid station, it is more efficient than in the past. And a stable shochu production system can be provided.

Brief description of the drawings

BRIEF DESCRIPTION OF THE DRAWINGS It is a figure which shows the relationship between the usage amount of the string in the domestic bacteria culture | cultivation using the liquid medium which used the crude barley, and the production | generation amount of glucoamylase and acid-resistant alpha-amylase.

Fig. 2 is a diagram showing the relationship between the amount of circumference of vein in domestic culture using a liquid medium using a polished barley of a control group, and the amount of glucoamylase and acid-resistant α-amylase produced.

It is a figure which shows the progress of fermentation in the production of barley shochu by the liquid station which used string wheat.

It is a figure which shows the fermentation process in the rice shochu production by the liquid soup which used buckwheat.

It is a figure which shows the fermentation progress in rice shochu production by the liquid station which used the tank.

It is a figure which shows the fermentation progress in rice shochu production by the liquid soup which used blood.

It is a figure which shows the fermentation progress in the manufacture of rice shochu by the liquid station which used millet.

Fig. 8 is a diagram showing the fermentation process in the production of rice shochu by the liquid soup using a large amount.

9 is a diagram showing the progress of fermentation in the production of shochu using various stations.

It is a figure which shows the fermentation process in the preparation of the soju using the liquid station obtained by the bacterium culture using the liquid medium which used soybean.

It is a figure which shows the progress of the fermentation in the preparation of the soju obtained by the bacteria culture | cultivation using the liquid medium which used the adzuki bean.

Carrying out the invention  Best form for

Hereinafter, the present invention will be described in detail.

In the method for producing a liquid station according to the present invention, cultivation of the bacterium in a liquid medium prepared by adding raw materials such as cereals, beans, potatoes, and specific grains, and the enzyme of glucoamylase and acid-resistant α-amylase It includes a process for producing a liquid station with increased activity. That is, since the bacterium is cultured using the above various raw materials, saccharification of starch in the raw materials takes time, the release rate of sugar to the culture system is suppressed, and the enzyme activity of the liquid station is enhanced. Glucoamylase and acid resistant α-amylase are simultaneously produced and accumulated in a balanced manner.

In the present invention, grains used as raw materials may include barley, rice, wheat, buckwheat, blood, crude, millet, corn amount, corn, and the like. As the shape of these raw materials, a fine white matter, or at least a fine white ratio of whitened to the extent that grains remain on the surface of the grain can be used. For example, in the case of grains of barley, the percentage of whitening of 100% of white rice or the percentage of whitening of white rice is 100%, and the percentage of grain of barley (generally 7) Minus 8%), that is, above 92% to 93%.

Here, the whitening ratio refers to the ratio of the grains remaining by whitening the grains. For example, the whitening ratio 90% means to cut 10% of the grains of the surface layer portion of the grains. In addition, the wort in this invention includes the thing from the unrefined barley to the thing whitened to the extent that the grains remain on the surface of a grain, ie, the thing with 90% or more of whitening ratio. In addition, a grain is the thing of the outer side which covers the surface of grain of a grain.

In the present invention, brown rice, which is a grain obtained by removing only grains of the surface used as a raw material, removes only rice husks.

In the present invention, soybeans, red beans, sweet potatoes and the like can be cited as beans and potatoes used as raw materials. These raw materials are only for wiping off the contamination of the outer shell, and have not been processed at all, such as cutting and grinding.

In the present invention, amaranthus, which is used as a raw material, is a generic term for amaranth and amaranth plants, and has a high protein content in cereals, and the content of lysine, which is one of amino acids, is comparable to soybeans. In addition, as a highly nutritious grain containing more calcium, iron, and fiber than white rice, the country of origin is a specific region of Latin America, India, the Himalayas, and Nepal. Quinoa, on the other hand, is a year old in the Agatha family and is cultivated mainly in the highlands of southern Peru and the Andes in western Bolivia, and contains abundant minerals, vitamins, proteins, and plant fiber.

Amaranthus and quinoa of a raw material may be used individually, or may be used in combination. These are used for the preparation of a liquid medium without pretreatment such as grinding or crushing.

The above raw material is mixed with water to prepare a liquid medium. The blending ratio of the raw materials is selected in an appropriate amount so that the glucoamylase and the acid resistant α-amylase are selectively generated and accumulated during the culture of the domestic bacteria, to prepare a liquid medium.

For example, in the case of using barley as a raw material, it is prepared by a liquid medium in which 1 to 20% (w / vol) of wort is added to water. In the case where a white barley is used as the wort, more preferably 8 to 10% (w / vol) is added to the liquid medium, and more preferably 95% white barley is used as the raw material. Is prepared with a liquid medium added 1-4% (w / vol).

Subsequently, in the case of using brown rice without chaff as a raw material, brown rice is 1% (w / vol) to 20% (w / vol), preferably 5% (w / vol) to 13% (w / vol), more preferably 8% (w / vol) to 10% (w / vol).

When using legumes as raw materials, 1-10% (w / vol) legumes with respect to water, preferably 8-10% (w / vol) with soybeans and 1-2% (w / vol) is prepared with the added liquid medium. In addition, when potatoes are used as a raw material, they are prepared in a liquid medium in which 1 to 10% (w / vol) of potatoes is added to water.

For example, when amaranthus is used as a raw material, 1.5% (w / vol) to 15% (w / vol) with respect to water, preferably 2% (w / vol) to 10% (w) / vol), more preferably 2% (w / vol) to 8% (w / vol). On the other hand, in the case of quinoa, 1.5% (w / vol) to 7% (w / vol) to water, preferably 2% (w / vol) to 6% (w / vol), more preferably It is prepared with a liquid medium added with 2% (w / vol) to 4% (w / vol).

Thus, since the optimum compounding usage amount differs depending on the polishing degree of the raw material to be used, the national strain used, the kind of raw material, etc., it is good to select arbitrarily.

By culturing the bacterium in a liquid medium to which the appropriate amount of the raw material is added, the enzymes of glucoamylase and acid resistant α-amylase are produced in a balanced and high yield, and a liquid station having sufficient enzymatic activity for use in brewing shochu is obtained. When the amount of the raw material used exceeds the upper limit, the viscosity of the culture solution becomes high, the supply of oxygen and air necessary for aerobic culture of Korean bacteria is insufficient, the oxygen concentration in the culture decreases, and the culture proceeds. It is not desirable because it becomes difficult. On the other hand, when the usage-amount of the said raw material is less than a lower limit, the target enzyme will not be produced high.

The starch contained in the raw material may be gelatinized before culturing. There is no restriction | limiting in particular about the gelatinization method of starch, What is necessary is just to perform it by conventional methods, such as a steaming method and a roasting method. In the sterilization step of the liquid medium, which will be described later, in the case of heating at a temperature higher than the gelatinization temperature of the starch by high temperature and high pressure sterilization or the like, the gelatinization of the starch is simultaneously performed by this treatment.

It is preferable to add an organic substance, an inorganic salt, etc. to a liquid medium suitably as a nutrient source other than the raw material mentioned above. These additives are not particularly limited as long as they are generally used for cultivation of bacteria, but as organic matter, rice bran, wheat bran, corn steep liquor, soybean cake, skim soybean ( defatted soybean) and the like, and the inorganic salts include water-soluble compounds such as ammonium salt, nitrate salt, potassium salt, acidic phosphate salt, calcium salt and magnesium salt, and two or more kinds of organic substances and / or inorganic salts may be used simultaneously. The amount of these additions is not particularly limited as long as it promotes the growth of bacteria, but it is preferable to add about 0.1 to 5% (w / vol) as the organic material and about 0.1 to 1% (w / vol) as the inorganic salt. The liquid medium of the bacterium obtained in this manner may be sterilized as necessary, and the treatment method is not particularly limited. As an example, a high temperature high pressure sterilization method is mentioned and what is necessary is just to carry out at 121 degreeC for 15 minutes.

After cooling the sterilized liquid medium to the culture temperature, the bacteria are inoculated into the liquid medium. The bacterium used in the present invention is a bacterium having a glycolytic enzyme producing ability, preferably a bacterium having a glucoamylase producing ability and an acid resistant α-amylase producing ability, for example, Aspergillus kawachii (Aspergillus kawachii). Bacillus bacillus, Aspergillus niger, Aspergillus awamori, Aspergillus niger, etc. Aspergillus oryzae, aspergillus sojae (Aspergillus sojae) and the like are represented by the Kukguk (黄 麴菌). In addition, the form of the domestic bacteria inoculated into a medium is arbitrary, and spores or mycelium can be used.

These bacterium can be used either by culturing with one strain or mixed culture with two or more strains of the same or different strains. Although they do not have any problem even if they use any form of mycelium obtained by spores or preculture, the use of mycelium is preferable because the time required for logarithmic growth is shortened. There is no particular restriction on the amount of inoculation of Korean bacteria into the liquid medium.However, in the case of spores, about 1 × 10 ⁴ to 1 × 10 ⁶ per ml of liquid medium is inoculated, and about 0.1 to 10% of the preculture solution is used for mycelia. desirable.

The culture temperature of the bacterium is not particularly limited as long as it does not affect growth, but preferably 25 to 45 ° C, more preferably 30 to 40 ° C. When the incubation temperature is low, the growth of Korean bacteria is slowed, and contamination by various bacteria is likely to occur. Incubation time is preferably 24 to 72 hours. The culture apparatus may be any one capable of liquid culture, but since the bacteria are required to perform aerobic culture, it is necessary to carry out under aerobic conditions that can supply oxygen or air to the medium. In addition, it is preferable to stir so that the raw material, oxygen, and a domestic bacteria in a medium may be distributed uniformly in an apparatus during culturing. As for the stirring conditions and the aeration amount, any conditions may be used as long as the culture environment can be maintained aerobicly, and may be appropriately selected depending on the viscosity of the culture apparatus and the medium.

By culturing in the above-mentioned culture method, enzymes of glucoamylase and acid resistant α-amylase are simultaneously produced in a balanced manner and become a liquid station having enzymatic activity that can be used for brewing shochu. In addition to the culture itself, the liquid station obtained by the above culture method may be a culture solution obtained by centrifugation or the like, a concentrate thereof, or a dried product thereof.

The liquid station obtained by the manufacturing method of this invention, etc. can be used for manufacture of liquor or a fermented food-drink. For example, in the case of making sake, in the sake brewing or in each mashing preparing step, in the case of making soju, in the mash soaking step, in the case of making soy sauce, piling In the) step, in the case of making miso, in the mashing step, in the case of preparing sweet sake, in the preparing step, a liquid station or the like may be used instead of the solid station.

In the case of producing liquor or fermented food or drink using the culture broth obtained from the liquid station or culture described above, concentrates thereof, or the like, the whole process can be performed in the liquid phase. As a method for producing liquor that performs the whole process in a liquid phase, for example, when preparing soju, corn, barley, rice, potato, sugar cane, etc. are used as an additive raw material, and the raw material is heat-resistant at a high temperature of about 80 ° C. After melting and liquefying using an enzyme agent, a method of distilling the mash subjected to alcohol fermentation by adding the above-mentioned liquid soup and yeast to distillation by atmospheric distillation or vacuum distillation may be used. .

Hereinafter, although an Example etc. demonstrate this invention more concretely, this invention is not limited to these Examples.

<Experimental example 1> [Investigation of the use amount of string in manufacturing a liquid station]

Five types of liquid media were prepared by changing the wort ratio of the raw materials as shown in Table 1, and cultured the bacterium in each liquid media to prepare a liquid station.

First, 5% of potassium nitrate and 0.3% potassium dihydrogen phosphate (w / vol) were added so that the string was 1, 2, 4, 8, 10% (w / vol). A kind of liquid medium was prepared. For each liquid medium, 100 ml of the prepared liquid medium was placed in a baffle flask with a volume of 500 ml, sterilized by autoclave, and pre-cultured in liquid medium (Aspergillus kawachii IFO4308). Was inoculated to 1% (v / vol) relative to the liquid medium. In addition, the string was used as a fine white of Japanese two-rowed barley (barley).

Then, the culture was performed for 48 hours at a temperature of 37 ° C. and a shaking speed of 100 rpm. After the completion of the culture, the amount of production of glucoamylase and acid resistant α-amylase was measured for each of the obtained cultures. In addition, Table 1 and Figure 1 shows the production amount of glucoamylase and acid resistant α-amylase of the culture obtained by culturing in liquid medium for each usage of the string. In addition, the saccharification power fractional quantification kit (made by Gigcommann) was used for the measurement of the enzyme activity of a glucoamylase. In addition, the measurement of the enzyme activity of acid-resistant alpha-amylase is a slight improvement of the method described in <Patent Document 3>, and after inactivating a non-acid-resistant alpha-amylase by acid-treating a culture, the alpha-amylase measurement kit ( Acid-resistant α-amylase was measured using Gikcommann. More specifically, 9 ml of 100 mM acetic acid buffer solution (pH 3) was added to 1 ml of the culture solution, and acid treatment was performed at 37 ° C. for 1 hour, followed by measurement using an α-amylase measurement kit (manufactured by Gikkoman).

On the other hand, as a control, a liquid medium was prepared in the same manner as the experimental group using barley (hereinafter referred to as circular vein) in which raw Japanese barley barley was whitened to a raw material ratio of 70% as a raw material, under the same conditions. After the completion of the culture, the amount of production of glucoamylase and acid resistant α-amylase was measured for each of the obtained cultures. Table 1 and Figure 2 show the glucoamylase and acid-resistant α-amylase production of the culture obtained by culturing in the liquid medium for each use of circular vein.

As shown in Table 1 and FIG. 1, the cultivation using the wort showed that the enzymes of glucoamylase and acid-resistant α-amylase were simultaneously balanced and high-produced with the increase in the amount of the wort used, and the yield was also used in the control vein of the control. It was confirmed that the increase significantly compared with that. In particular, 217.3 U / ml of glucoamylase and 14.0 U / ml of acid-resistant α-amylase were produced in the liquid medium to which 10% (w / vol) of wort was added, so that sufficient enzyme activity for use in brewing shochu was simultaneously obtained. (For reference, the enzyme activity values of glucoamylase and acid-resistant α-amylase required for the preparation of soju are at least 100 U / ml for glucoamylase and at least 10 U / ml for acid-resistant α-amylase.).

On the other hand, in the control group using circular vein, as shown in Table 1 and FIG. 2, the glucoamylase activity was maximum in the liquid medium to which 2% (w / vol) of circular vein was added, and acid-resistant α-amylase activity was 8% ( w / vol) in the added liquid medium, but both enzymes were not produced at the same time.

In this way, glucoamylase and acid resistant α-amylase are simultaneously produced in good balance by culturing the bacterium in a liquid medium to which 1 ~ 20% (w / vol) of wort is added. In particular, addition of 8 to 10% (w / vol) of wort (white rice) enables the production of a liquid station in which sufficient enzyme activity is simultaneously obtained for use in soju brewing.

When cultivation of Korean bacteria in a liquid medium using a string of wheat, glucoamylase and acid-resistant α-amylase are simultaneously produced in a well-balanced manner. Since it is suppressed by this grain and is culture | cultivated in the state where sugar concentration in culture is comparatively low, it is thought that it is because it is easy to produce enzymes, such as glucoamylase and acid-resistant alpha-amylase.

Example 1 Preparation of Liquid Station Using Strings

First, a liquid medium was added so that the string was 10% (w / vol) in water to which 0.2% (w / vol) of potassium nitrate and 0.3% (w / vol) of potassium dihydrogen phosphate were added. Subsequently, 100 ml of the prepared liquid medium was placed in a 500 ml baffle conical flask, and after autoclave sterilization, 1% (v / vol) of the Bacillus bacterium (Aspergillus kawachii IFO4308) pre-incubated in the liquid medium was added to the liquid medium. ) Was inoculated. In addition, the string used the thing which is a white of Japanese two lined barley.

Then, the culture was performed for 48 hours at a temperature of 37 ° C. and a shaking speed of 100 rpm. After the end of the culture, the amount of glucoamylase and acid resistant α-amylase produced was measured for each of the cultures obtained. As a result, glucoamylase produced 217.3 U / ml and acid resistant α-amylase 14.0 U / ml. Enzyme activity sufficient for use was obtained simultaneously.

Example 2 Preparation of Barley Shochu by Liquid Soup with Strings

In Example 1, soju was prepared using a liquid station obtained by culturing in a liquid medium prepared by adding 10% (w / vol) of strings (a culture enhanced with glucoamylase and acid resistant α-amylase).

That is, the total barley 1328.6 was obtained as a mashing combination shown in Table 2 using 500 ml of the liquid broth obtained by culturing in a liquid medium prepared by adding 10% (w / vol) of string in Example 1. g soaking was performed, and the fermentation temperature was maintained at 25 ° C., and three stages of immersion were performed for 5 days of primary immersion, 2 days of secondary immersion, and 13 days of 3rd immersion. In addition, as the additive barley, 70% white barley of Japanese barley was washed with water, immersed in water for 60 minutes, drained for 30 minutes, and steamed for 35 minutes. In addition, in primary immersion, since the amount of 50.0 g of barley taken from the liquid state is insufficient for fermentation, 262.9 g of mixed barley was immersed so that the same amount of barley as the solid state immersion entered. In addition, the yeast was 50 microliters culture | cultivated using the soju yeast (Kagoshima yeast), and statically incubated for 48 hours at 30 degreeC in YPD medium.

Moreover, as a control immersion (solid station immersion), the soju was produced by the immersion formulation shown in Table 3 using the burley of the solid station. In the production method of the solid state, using 70% white barley, washing the barley, immersing for 40 minutes, draining for 30 minutes, steaming for 40 minutes, and cooling down to 40 ° C, 1 kg of white barley 1 g of the final product (aspergillus Aspergillus kawachii IFO4308) was inoculated per 24 hours at 40 ° C and 95% RH, 6 hours at 35 ° C and 95% RH, and 30 ° C and 90% RH, respectively. Incubated for 18 hours. The fermentation conditions and the like were the same as those of the above-described immersion (liquid station immersion).

Its fermentation progress is shown in FIG. 3 as compared to the solid station dip of the control. As is apparent from FIG. 3, the fermentation process using the liquid station was almost the same as the control soaking using the solid station. In addition, the alcohol content of the obtained final mash was also the same at 17.8% using either a liquid station or a solid station.

Subsequently, the final mash obtained by distillation under reduced pressure was adjusted to 25% alcohol content by sensory evaluation by a panelist scoring method (five-point evaluation method, 1: good to 5: poor), Its average point is shown in Table 4.

As a result, the difference in shochu quality was hardly recognized, and it was confirmed that even if a liquid station was used, the production of shochu of the same quality as in the case of using a solid station was possible.

From the above results, according to the present invention, glucoamylase and acid resistant α-amylase are simultaneously produced in good balance by culturing the bacterium in a liquid medium containing 1 to 20% (w / vol) of wort. In particular, the addition of 8 to 10% (w / vol) of wort (white rice) enables the production of a liquid station in which sufficient enzyme activity is simultaneously obtained for use in soju brewing. For this reason, by using this liquid station, it becomes possible to manufacture the soju of the quality equivalent to the soju manufactured using the solid station. Moreover, a liquid station with high glucoamylase activity and acid resistance α-amylase activity can be produced in a simple liquid medium without strict culture control by a special culture device or a special culture engineering technique, and is also very rigid compared to solid culture. By easily performing the station manufacturing management, stable production of a station of high quality is possible. Furthermore, the liquidification of the soup makes it possible not only to facilitate fermentation management by fluidizing the mash, but also to save and efficiency of the soup manufacturing process, furthermore, the soju manufacturing process.

Example 3 Preparation of Rice Shochu with Liquid Bureau Using Buckwheat

1. Manufacturing method of solid state

After washing rice using 90% white rice, immersed for 15 minutes, drained for 10 minutes, steamed for 30 minutes, cooled to 40 ° C, and 1 g of final product (kg Aspergillus kawachii IFO4308) per kg of white rice was inoculated. The cells were incubated for 24 hours at 40 ° C and 95% RH, for 6 hours at 35 ° C and 95% RH, and for 18 hours at 30 ° C and 90% RH.

2. Liquid station preparation

(1) preculture method; 8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 40 g of buckwheat, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water were charged into a 2000 ml baffle conical flask, followed by autoclave sterilization at 121 ° C for 15 minutes. The buckwheat liquid state was produced by inoculating 5 ml of preculture liquids with this main culture medium, and shaking-culturing at 37 degreeC, 48 hours, and 100 rpm. The enzymatic activity of the liquid station at this time was GA activity 112.4 U / mL and ASAA activity 10.4 U / mL.

3. Rice shochu manufacturing method

(1) use yeast; Shochu yeast (Kagoshima yeast)

(2) immersion formulations; Dipping formulations are shown in Tables 5 and 6. 90% rice was used for rice, and after washing, the rice was immersed for 15 minutes, drained for 10 minutes, and steamed for 30 minutes. The test zones were 2 test zones of 1) solid state immersion and 2) buckwheat liquid state immersion, and the total rice and water supply amount of both test spheres were mixed so as to be the same amount. Yeast was incubated in 50 µl of the culture in a stationary culture at 30 ℃, 48 hours in YPD medium.

(3) fermentation conditions; 25 ℃ schedule

(4) distillation conditions; Vacuum distillation

The fermentation progress is shown in FIG. 4 as compared to the solid station immersion of the control. As is apparent from Fig. 4, the fermentation process using the buckwheat liquid station was substantially the same as the control immersion using the solid station. In addition, the alcohol counts of the final mash of the obtained solid-state mashing group and the buckwheat-liquid stationary immersion were 19.1% and 18.9%, respectively, and were about the same.

Subsequently, the sensory evaluation of the soju circumference produced by distilling the soju mash of the obtained solid state immersion sphere and the buckwheat liquid state immersion sphere by vacuum distillation was performed by a five-point evaluation method (good 1-3-5 poor) of six panelists. It was found that there is no significant difference between the solid state soaking and the buckwheat liquid soaking sauces, and even soba can be produced with sufficient quality of soju. Moreover, as shown in Table 7, in the buckwheat liquid soup district, there was also a comment "tastes of buckwheat", and it was confirmed that the use of the buckwheat liquid soup can impart "buckwheat" to rice shochu. .

Example 4 Preparation of Rice Shochu with Liquid Soup Using Bath

1. Manufacturing method of solid state

After washing rice using 90% white rice, immersed for 15 minutes, drained for 10 minutes, steamed for 30 minutes, cooled to 40 ° C, and 1 g of final product (kg Aspergillus kawachii IFO4308) per kg of white rice was inoculated. The cells were incubated for 24 hours at 40 ° C and 95% RH, for 6 hours at 35 ° C and 95% RH, and for 18 hours at 30 ° C and 90% RH.

2. Liquid station preparation

(1) preculture method; 8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 40 g of the crude tank, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water were charged into a 2000 ml baffle conical flask, followed by autoclave sterilization at 121 ° C for 15 minutes. 5 ml of the preculture was inoculated with this main culture medium, and a crude liquid station was prepared by shaking culture at 37 ° C for 48 hours at 100 rpm. Enzyme activity of the liquid station at this time was GA activity 101.3 U / mL and ASAA activity 11.0 U / mL.

3. Rice shochu manufacturing method

(1) use yeast; Shochu yeast (Kagoshima yeast)

(2) immersion formulations; Dipping formulations are shown in Table 8 and Table 9. 90% rice was used for rice, and after washing the rice, it was immersed for 15 minutes, drained for 10 minutes, and used for 30 minutes. The test zones were 2 test zones of 1) solid station immersion and 2) crude liquid station immersion, and the total rice and water supply amount of both test spheres were blended so as to be the same amount. Yeast was incubated in 50 µl of the culture in a stationary culture at 30 ℃, 48 hours in YPD medium.

(3) fermentation conditions; 25 ℃ schedule

(4) distillation conditions; Vacuum distillation

The fermentation progress is shown in FIG. 5 as compared to the solid station immersion of the control. As is apparent from FIG. 5, the fermentation process using the crude liquid station showed almost the same fermentation progress as compared to the control soaking using the solid state. In addition, the alcohol content of the final mash of the obtained solid-station quenching apparatus and the crude liquid-station quenching apparatus was all the same at 19.1%.

Subsequently, the sensory evaluation of the soju circumference produced by distilling the soju mash in the solid state immersion and the crude liquid state immersion by the distillation under reduced pressure distillation was carried out by a five-point evaluation method of six expert panelists (good 1-3-5 poor). It was found that there is no big difference between the soaking soup and the soaking soup, and even a soaking soup can be made of soju of sufficient quality. Furthermore, as shown in Table 10, there was also a comment that the crude liquid soup was "clear tastes," suggesting the possibility of producing a rice wine liquor of a quality different from the conventional solid soup manufacturing method.

Example 5 Preparation of Rice Shochu with Liquid Soup Using Blood

1. Manufacturing method of solid state

After washing rice using 90% white rice, immersed for 15 minutes, drained for 10 minutes, steamed for 30 minutes, cooled to 40 ° C, and 1 g of final product (kg Aspergillus kawachii IFO4308) per kg of white rice was inoculated. The cells were incubated for 24 hours at 40 ° C and 95% RH, for 6 hours at 35 ° C and 95% RH, and for 18 hours at 30 ° C and 90% RH.

2. Liquid station preparation

(1) preculture method; 8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 40 g of blood, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water were charged into a 2000 ml baffle flask, and autoclave sterilized for 15 minutes at 121 ° C. 5 mL of the preculture solution was inoculated with this main culture medium, and the blood station was prepared by shaking culture at 37 ° C for 48 hours at 100 rpm. The enzymatic activity of the liquid station at this time was GA activity 113.0 U / mL and ASAA activity 10.2 U / mL.

3. Rice shochu manufacturing method

(1) use yeast; Shochu yeast (Kagoshima yeast)

(2) immersion formulations; Dipping formulations are shown in Tables 11 and 12. 90% rice was used for rice, and after washing the rice, it was immersed for 15 minutes, drained for 10 minutes, and used for 30 minutes. The test zones were 2 test zones of 1) solid station immersion and 2) liquid station immersion, and the total rice and water supply of both test spheres were blended so as to be the same amount. Yeast was incubated in 50 µl of the culture in a stationary culture at 30 ℃, 48 hours in YPD medium.

(3) fermentation conditions; 25 ℃ schedule

(4) distillation conditions; Vacuum distillation

The fermentation progress is shown in FIG. 6 as compared to the solid station immersion of the control. As is apparent from FIG. 6, the fermentation process using the liquid station showed almost the same fermentation progress as compared with the control soaking using the solid station. In addition, the alcohol content of the final mash of the obtained solid-state immersion tool and the liquid station immersion tool was 19.1% and 18.8%, respectively, about the same grade.

Subsequently, the sensory evaluation of the soju circumference produced by distilling the soju mash in the solid state immersion sphere and the liquid state immersion sphere by vacuum distillation was carried out by a five-point evaluation method (good 1-3-5 poor) of six panelists. It was found that there is no significant difference between the soaking soup and the soaking soup, and even the soaking soup can be produced with sufficient quality. Moreover, as shown in Table 13, there was also a comment that the liquid soup ball was "neat taste", suggesting the possibility of producing rice shochu circumference of a quality different from the conventional solid soup production method.

Example 6 Preparation of Rice Shochu with Liquid Soup Using Millet

1. Manufacturing method of solid state

After washing rice using 90% white rice, immersed for 15 minutes, drained for 10 minutes, steamed for 30 minutes, cooled to 40 ° C, and 1 g of final product (kg Aspergillus kawachii IFO4308) per kg of white rice was inoculated. The cells were incubated for 24 hours at 40 ° C and 95% RH, for 6 hours at 35 ° C and 95% RH, and for 18 hours at 30 ° C and 90% RH.

2. Liquid station preparation

(1) preculture method; 8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 40 g of millet, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water were charged into a 2000 ml baffle conical flask, followed by autoclave sterilization at 121 ° C for 15 minutes. 5 ml of the preculture was inoculated with this main culture medium, and the milled liquid station was prepared by shaking culture at 37 ° C for 48 hours at 100 rpm. At this time, the enzyme activities of the liquid station were GA activity 90.3 U / ml and ASAA activity 8.5 U / ml.

3. Rice shochu manufacturing method

(1) use yeast; Shochu yeast (Kagoshima yeast)

(2) immersion formulations; Dipping formulations are shown in Table 14 and Table 15. 90% rice was used for rice, and after washing the rice, it was immersed for 15 minutes, drained for 10 minutes, and used for 30 minutes. The test zones were 2 test zones of 1) solid station immersion and 2) millet liquid station immersion, and the total amount of rice and water supply of both test spheres were blended to be the same amount. Yeast was incubated in 50 µl of the culture in a stationary culture at 30 ℃, 48 hours in YPD medium.

(3) fermentation conditions; 25 ℃ schedule

(4) distillation conditions; Vacuum distillation

The fermentation progress is shown in FIG. 7 as compared to the solid station dip of the control. As is apparent from FIG. 7, the fermentation using the milled liquid station was almost the same as the control immersion using the solid station. In addition, the alcohol content of the final mash of the obtained solid-state immersion tool and millet liquid-station immersion tool was 19.1% and 18.8%, respectively, about the same grade.

Subsequently, the sensory evaluation of the soju circumference produced by distilling the soju mash in the solid state immersion sphere and the millet liquid state immersion sphere by vacuum distillation was performed by a five-point evaluation method (good 1-3-5 poor) of six panelists. It was found that there is no big difference between the solid state soak and the millet liquid station soak, and even the milled liquid station can produce sake of sufficient quality. Furthermore, as shown in Table 16, there was also a comment that the milled liquid soup ball had "mild and sweet tastes," suggesting the possibility of producing rice shochu spirits of a quality different from the conventional solid soup method. .

<Example 7> [Preparation of Rice Shochu by Liquid Soup Using High Amount]

1. Manufacturing method of solid state

After washing rice using 90% white rice, immersed for 15 minutes, drained for 10 minutes, steamed for 30 minutes, cooled to 40 ° C, and 1 g of final product (kg Aspergillus kawachii IFO4308) per kg of white rice was inoculated. The cells were incubated for 24 hours at 40 ° C and 95% RH, for 6 hours at 35 ° C and 95% RH, and for 18 hours at 30 ° C and 90% RH.

2. Liquid station preparation

(1) preculture method; 8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 40 g of a large amount, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water were charged into a 2000 ml baffle conical flask, followed by autoclave sterilization at 121 ° C for 15 minutes. 5 ml of the preculture was inoculated with this main culture medium, and a high amount of liquid station was prepared by shaking culture at 37 ° C for 48 hours at 100 rpm. The enzymatic activity of the liquid station at this time was GA activity 111.2 U / mL and ASAA activity 10.5 U / mL.

3. Rice shochu manufacturing method

(1) use yeast; Shochu yeast (Kagoshima yeast)

(2) immersion formulations; Dipping formulations are shown in Table 17 and Table 18. 90% rice was used for rice, and after washing the rice, it was immersed for 15 minutes, drained for 10 minutes, and used for 30 minutes. The test zones were 2 test zones of 1) solid station immersion and 2) high-volume liquid station immersion, and the total rice and water supply amount of both test spheres were blended so as to be the same amount. Yeast was incubated in 50 µl of the culture in a stationary culture at 30 ℃, 48 hours in YPD medium.

(3) fermentation conditions; 25 ℃ schedule

(4) distillation conditions; Vacuum distillation

The fermentation progress is shown in FIG. 8 as compared to the solid station immersion of the control. As is apparent from FIG. 8, almost the same fermentation process was exhibited in the immersion using a high amount of liquid station as compared to the control immersion using a solid station. In addition, the alcohol content of the final mash of the obtained solid-state immersion tool and the high-volume liquid-station quench both was the same at 19.1%.

Subsequently, the sensory evaluation of the soju circumference produced by distilling the soju mash in the solid state immersion sphere and the high-volume liquid state immersion sphere by vacuum distillation was carried out by a five-point evaluation method (good 1-3-5 poor) of six panelists. It was found that there is no big difference between the soak soup and the high liquid soak soup, and even a high soak liquid soup can produce sake of sufficient quality. Furthermore, as shown in Table 19, there is also a comment that the high-volume liquid soup has a "cereal-like sweetness," suggesting the possibility of making a distilled liquor that is distinctly different from the conventional solid soup manufacturing method. .

Example 8 Preparation of Liquid Station Using Corn

(1) preculture method; 8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 1-8 g of corn, 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized for 15 minutes at 121 ° C. 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm. The production amount of the enzyme in the culture supernatant after the culture, that is, the glucoamylase (GA) activity and the acid resistance α-amylase (ASAA) activity was measured by the method described in Experimental Example 1. The results are shown in Table 20.

As is apparent from Table 20, in a test plot of 4% or more corn usage, 100 U / ml of glucoamylase, which is a target of enzymatic activity required for brewing soju, was achieved. On the other hand, the target value of the acid-resistant α-amylase was 10 U / ml, but this target value was not reached, but as the amount of corn used increased, the production of the enzyme was confirmed to increase.

As mentioned above, although the target value of ASAA activity was not able to be achieved in this test, since it showed the ability to produce GA enzyme and ASAA enzyme simultaneously, the possibility of raising enzyme productivity to the target value level by the optimization of culture conditions is high. . In addition, for example, in the soju dipping using 8% corn liquid soup, it was inferred that it is possible to manufacture soju sufficiently if the soup ratio is increased more than normal mixing.

Example 9 Preparation of Liquid Station Using Strings

(1) preculture method; 8 g of 65% white rice and 100 ml of water were charged into a 500 ml baffle conical flask and autoclaved sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Aspergillus awamori IFO4388 were seeded in 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 1-8 g of string (95% white barley), 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm. The production amount of the enzyme in the culture supernatant after the culture, that is, the glucoamylase (GA) activity and the acid resistance α-amylase (ASAA) activity was measured by the method described in Experimental Example 1. The results are shown in Table 21.

As is apparent from Table 21, in the test plot of 4% of the amount of use of the wort, 100 U / ml of glucoamylase and 10 U / ml of acid resistant α-amylase, which are targets of enzymatic activity required for brewing soju, were simultaneously achieved. From this, it was confirmed that the high enzyme production effect appeared in the same manner as in the case of Baekkukyun even when using the Baekkukyun.

Example 10 Preparation of Liquid Soup Using Brown Rice (rice with chaff)

(1) preculture method; 8 g of 90% white rice (rice for eating) and 100 ml of water were charged into a 500 ml baffle conical flask and autoclave sterilized at 121 ° C. for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 1-8 g of brown rice (with chaff), 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were charged into a 500 ml baffle conical flask and sterilized at 121 ° C for 15 minutes. In addition, the brown rice used was used without threshing the thing with the grain (chaff) attached. 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm. The production amount of the enzyme in the culture supernatant after the culture, that is, the glucoamylase (GA) activity and the acid resistance α-amylase (ASAA) activity was measured by the method described in Experimental Example 1. The results are shown in Table 22.

As is apparent from Table 22, in the test plot of 4% of the brown rice consumption, 100 U / ml of glucoamylase and 10 U / ml of acid resistant α-amylase, which were targets of enzymatic activity required for brewing soju, were simultaneously achieved. From this, the enzyme high production effect was confirmed also about rice with grain (rice husk) similarly to string rice.

Example 11 Preparation of Liquid Soup Using Brown Rice (rice with chaff)

(1) preculture method; 8 g of 90% white rice (rice) and 100 ml of water were charged into a 500 ml baffle conical flask and autoclave sterilized for 15 minutes at 121 占 폚. After allowing to cool, the preculture medium was inoculated with 1 × 10 ⁶ spores of final spores of Aspergillus awamori IFO4388, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) the present culture method; 1-8 g of brown rice (with chaff), 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were charged into a 500 ml baffle conical flask and sterilized at 121 ° C for 15 minutes. In addition, the brown rice used was used, without threshing the thing with the grain of rice (rice husk). 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm. The production amount of the enzyme in the culture supernatant after the culture, that is, the glucoamylase (GA) activity and the acid resistance α-amylase (ASAA) activity was measured by the method described in Experimental Example 1. The results are shown in Table 23.

As is apparent from Table 23, in the test plot of 8% of the brown rice consumed, 100 U / ml of glucoamylase and 10 U / ml of acid resistant α-amylase, which are targets of enzymatic activity required for brewing soju, were simultaneously achieved. From this, the enzyme high production effect was confirmed even if the rice with grains (chaff) was used, and the black bacterium was used.

<Example 12> [Preparation of liquid station using brown rice without rice husk]

1. Pre-culture

8 g of 90% polished rice and 100 ml of water were charged into a 500 ml baffle conical flask and autoclaved sterilized at 121 ° C for 15 minutes. After cooling, the preculture medium was inoculated with final spores of Bacillus agar (Aspergillus kawachii IFO4308) to 1 × 10 ⁶ cells / ml, and cultured at 37 ° C. for 24 hours at 100 rpm.

2. Main culture method

1-10 g of rice with rice hulls removed, 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate and 100 ml of water were charged into a 500 ml baffle conical flask and autoclaved at 121 ° C. for 15 minutes. After cooling, 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm.

On the other hand, as a control, 1-10 g of 90% polished rice, 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were charged into a 500 ml baffle conical flask and autoclaved sterilized at 121 ° C for 15 minutes. After cooling, 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm.

After the completion of the culture, glucoamylase activity (GA) and acid resistant α-amylase (ASAA) in each culture supernatant were measured. The measurement of glucoamylase activity (GA) is performed using a glycosylation ability fractionation quantitative kit (Geek Coman), and the measurement of acid-resistant (alpha) -amylase activity (ASAA) slightly improves the method as described in <nonpatent literature 3>, Non-acid-resistant α-amylase activity was inactivated by acid treatment of the culture, followed by the α-amylase measurement kit (manufactured by Gigcommann). More specifically, 9 ml of 100 mM acetic acid buffer solution (pH 3) was added to 1 ml of the culture solution, and acid treatment was performed at 37 ° C. for 1 hour, followed by measurement using an α-amylase measurement kit (manufactured by Gikkoman).

3. Results

As shown in Table 24. Enzyme activity required for brewing shochu from the above studies is sufficient if it is about 100 U / mL of glucoamylase and about 10 U / mL of acid-resistant alpha-amylase. When the white rice of the control was used, GA and ASAA did not exceed the target at the same time, but when the brown rice of the test was used, both GA and ASAA tended to be produced in a balanced manner, and in particular, the target enzyme was used with 8% or more of brown rice. Activity values were achieved. From this result, it was shown that brown rice is suitable as a raw material of a liquid state compared with white rice.

Example 13 Preparation of Shochu by Liquid Soup Using Brown Rice from Rice Husk

1. Manufacturing method of solid state

After washing rice using 90% white rice, immersed for 15 minutes, drained for 10 minutes, steamed for 30 minutes, allowed to cool to 40 ° C, and 1 g of final product (kg Aspergillus kawachii IFO4308) per kg of white rice was inoculated. Then, the cultures were incubated for 18 hours at 40 ° C and 95% RH for 6 hours, and at 30 ° C and 90% RH.

2. Liquid station preparation

(1) Pre-cultivation method

8 g of 90% polished rice and 100 ml of water were charged into a 500 ml baffle conical flask and autoclaved sterilized at 121 ° C for 15 minutes. After cooling, the preculture medium was inoculated with final spores of Bacillus agar (Aspergillus kawachii IFO4308) to 1 × 10 ⁶ cells / ml, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) main culture method

40 g of rice with rice hulls removed, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water were charged into a 2000 ml baffle conical flask and autoclaved at 121 ° C. for 15 minutes. 5 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm to prepare a brown rice liquid soup.

As a control, 40 g of 90% polished rice, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate and 500 ml of water were charged into a 2000 ml baffle conical flask and autoclaved sterilized at 121 ° C. for 15 minutes. 5 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm to prepare a white rice liquid soup.

3. Rice shochu manufacturing method

(1) use yeast; Shochu yeast (Kagoshima yeast)

(2) immersion formulation

Dipping formulations are as shown in Tables 25-27. 90% rice was used for rice, and after washing the rice, it was immersed for 15 minutes, drained for 10 minutes, and used for 30 minutes. The experimental zones (test zone, control zone) were three spheres of 1) solid soup immersion, 2) brown rice liquid soup immersion, and 3) white rice liquid soup immersion, and the total rice and water supply of each ball were blended so as to be the same amount. Yeast was incubated in 50 µl of the culture in a stationary culture at 30 ℃, 48 hours in YPD medium.

(3) fermentation conditions; 25 ℃ schedule

(4) distillation conditions; Vacuum distillation

4. Results and Discussion

The fermentation progress is as shown in FIG. As is apparent from the figure, the solid-state immersion sphere and the brown rice liquid-state immersion sphere showed almost the same fermentation progress. However, the fermentation process of the white rice liquid soup soaking was inferior. In addition, the alcohol content of the final mash obtained was 19.1% of the solid state immersion sphere and the 18.9% of the brown rice liquid state immersion sphere, and the alcohol frequency of the final mash of the white rice liquid state immersion sphere was 12.5%. It is greatly under).

The sensory evaluation of the distilled liquor made by distillation of the soju mash in the solid soak soup and brown rice liquid soup soak by the distillation under reduced pressure distillation was performed by a five-point evaluation method (good 1-3-5 bad) of six panelists. As shown in the figure, there was no significant difference between the solid station immersion and the brown rice liquid station. However, there was a comment that the brown rice liquid soup was "clear and refreshing", and it was found that rice shochu with a refreshing flavor was obtained. From the above result, it was shown that the rice wine liquor of the quality equivalent to a solid state can also be produced by a brown rice liquid state.

Example 14 Preparation of Liquid Soup Using Brown Rice from Rice Husk

1. Pre-culture

8 g of 90% white rice (rice) and 100 ml of water were charged to a 500 ml baffle conical flask and autoclaved at 121 ° C for 15 minutes. After allowing to cool, the preculture medium was inoculated with 1 × 10 ⁶ spores of final spores of Aspergillus awamori IFO4388, and cultured at 37 ° C. for 24 hours at 100 rpm.

2. Main culture method

1-8 g of rice with rice husks removed, 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were charged into a 500 ml baffle conical flask and autoclaved at 121 ° C. for 15 minutes. After cooling, 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm.

After the completion of the culture, glucoamylase activity (GA) and acid resistant α-amylase (ASAA) in the culture supernatant were measured by the method described in Example 12.

3. Results

As shown in Table 29. As mentioned above, it is considered that the target value of the enzyme activity necessary for brewing shochu is about 100 U / mL of glucoamylase and about 10 U / mL of acid resistant α-amylase. As is apparent from Table 29, it was confirmed that in the test plot of 8% brown rice consumption, both GA and ASAA activities achieved the target values, and even if the black bacillus bacterium was used, the high enzyme production effect appeared as in the case of the Bacillus bacillus. . In addition, even when the amount of brown rice used is further vaporized, high enzyme production effects can be expected in the same manner as Baekkukyun.

Example 15 Preparation of Soybean Liquid Soup

1. Pre-culture

8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, the preculture medium was inoculated with final spores of Bacillus agar (Aspergillus kawachii IFO4308) to 1 × 10 ⁶ cells / ml, and cultured at 37 ° C. for 24 hours at 100 rpm.

2. Main culture method

1-10 g of soybeans, 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were charged into a 500 ml baffle conical flask, followed by autoclave sterilization at 121 ° C. for 15 minutes. 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm. After incubation, the activity of glucoamylase (GA) and acid resistant α-amylase (ASAA) in the culture supernatant was measured. Measurement of GA activity is performed using a glycosylation fractionation quantitative kit (manufactured by Gikkoman), and measurement of ASAA activity is slightly improved by the method described in <Non-Patent Document 3>, and the acid-treated culture of the non-acid-resistant α- After inactivation of amylase activity, it was carried out using the α-amylase measurement kit (Gikkomann). More specifically, 9 ml of 100 mM acetic acid buffer solution (pH 3) was added to 1 ml of the culture solution, and acid treatment was performed at 37 ° C. for 1 hour, followed by measurement using an α-amylase measurement kit (manufactured by Gikkoman).

3. Results

As shown in Table 30. The target values of the enzyme activity required for the preparation of soju in the above studies are 100 U / ml for glucoamylase activity and 10 U / ml for acid resistant α-amylase activity. As is apparent from the table, GA activity and ASAA activity also increased with increasing amount of soybean, and target enzyme activity was achieved with the use of 8% or more soybean. From this result, it was shown that soybean is suitable as a raw material of the liquid station.

Example 16 Preparation of Rice Shochu Using Soybean Liquid Soup

1. Manufacturing method of solid state

After washing rice using 90% white rice, immersed for 15 minutes, drained for 10 minutes, steamed for 30 minutes, cooled to 40 ° C, and 1 g of final product (kg Aspergillus kawachii IFO4308) per kg of white rice was inoculated. The cells were incubated for 24 hours at 40 ° C and 95% RH, for 6 hours at 35 ° C and 95% RH, and for 18 hours at 30 ° C and 90% RH.

2. Liquid station preparation

(1) Pre-cultivation method

8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) main culture method

40 g of soybean, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water were charged into a 2000 ml baffle conical flask and autoclaved at 121 ° C for 15 minutes. Soybean liquid soup was prepared by inoculating 5 ml of the pre-culture solution with this main culture medium and shaking culture at 37 ° C for 48 hours at 100 rpm.

3. Rice shochu manufacturing method

(1) use yeast; Shochu yeast (Kagoshima yeast)

(2) immersion formulation

Dipping formulations are as shown in Table 31 and Table 32. 90% pure rice was used as the rice, and after washing the rice, it was immersed for 15 minutes, drained for 10 minutes, and steamed for 30 minutes. The test zones were two test zones of (1) solid state immersion and (2) soybean liquid state immersion, and the total rice and water supply amount of both test spheres were blended so as to be the same amount. In addition, 50 μl of the yeast was cultured in a stationary culture at 30 ° C. for 48 hours in YPD medium.

(3) fermentation conditions; 25 ℃ schedule

(4) distillation conditions; Vacuum distillation

4. Results and Discussion

The fermentation progress is shown in FIG. As is apparent from the figure, the rice solid soup immersion sphere of the control and the soybean liquid soup immersion sphere of the test sphere showed almost the same fermentation progress. However, the alcohol content of the obtained final mash was the same as that of the rice solid soup immersion apparatus in 19.1% and the soybean liquid soup immersion apparatus in 18.7%.

The sensory evaluation of Suzhou brewed liquor made by distilling the soju mash of rice solid soup and soybean liquid soup by distillation under reduced pressure distillation was carried out by five panel evaluation method of six expert panelists (Good 1-3-5 bad). As shown in 33, there was no significant difference between the rice solid soup soaking and soybean liquid soup soaking, and it was found that even soybean liquid soup could be made of soju of sufficient quality. In addition, there was also a comment that the soybean liquid soup was "gorgeous flavors," suggesting the possibility of making a distilled liquor that is clearly different from the conventional solid state recipe.

Example 17 Preparation of Red Bean Liquid Soup

1. Pre-culture

8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, the preculture medium was inoculated with final spores of Bacillus agar (Aspergillus kawachii IFO4308) to 1 × 10 ⁶ cells / ml, and cultured at 37 ° C. for 24 hours at 100 rpm.

2. Main culture method

1-10 g of red beans, 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were filled into a 500 ml baffle conical flask, and autoclave sterilized for 15 minutes at 121 ° C. 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm. Glucoamylase (GA) activity and acid resistant α-amylase (ASAA) activity in the culture supernatant after culture were measured by the same method as in Example 15.

3. Results

As shown in Table 34. As mentioned above, the target value of the enzyme activity required for the preparation of soju is 100 U / ml for glucoamylase activity and 10 U / ml for acid resistant α-amylase activity. As is apparent from the table, the GA activity and ASAA activity were maximized by 2% red beans usage, and the target enzyme activity was achieved by 1-2% red beans usage. From this result, it was shown that adzuki beans are suitable as a raw material for liquid soup.

Example 18 Preparation of Rice Shochu with Red Bean Liquid Soup

1. Manufacturing method of solid state

After washing rice using 90% white rice, immersed for 15 minutes, drained for 10 minutes, steamed for 30 minutes, cooled to 40 ° C, and 1 g of final product (kg Aspergillus kawachii IFO4308) per kg of white rice was inoculated. The cells were incubated for 24 hours at 40 ° C and 95% RH, for 6 hours at 35 ° C and 95% RH, and for 18 hours at 30 ° C and 90% RH.

2. Liquid station preparation

(1) Pre-cultivation method

8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, seed culture spores of Bacillus agar (Aspergillus kawachii IFO4308) were inoculated to 1 × 10 ⁶ cells / ml in the pre-culture medium, and cultured at 37 ° C. for 24 hours at 100 rpm.

(2) main culture method

10 g of red beans, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water were filled into a 2000 ml baffle flask, followed by autoclave sterilization at 121 ° C for 15 minutes. The red bean liquid soup was produced by inoculating 5 ml of preculture liquids with this main culture medium, and shaking-culturing at 37 degreeC, 48 hours, and 100 rpm.

3. Rice shochu manufacturing method

(1) use yeast; Shochu yeast (Kagoshima yeast)

(2) immersion formulation

Dipping formulations are as shown in Table 35 and Table 36. 90% pure rice was used as the rice, and after washing the rice, it was immersed for 15 minutes, drained for 10 minutes, and steamed for 30 minutes. The test zones were two test zones of (1) solid state immersion sphere and (2) adzuki bean liquid state immersion, and the total rice and water supply amount of both test spheres were blended so as to be the same amount. In addition, 50 μl of the yeast was cultured in a stationary culture at 30 ° C. for 48 hours in YPD medium.

(3) fermentation conditions; 25 ℃ schedule

(4) distillation conditions; Vacuum distillation

4. Results and Discussion

The fermentation progress is shown in FIG. As is apparent from the figure, the rice solid soup soaking and the red bean liquid soup soaking showed almost the same fermentation progress. In addition, the alcohol content of the obtained final mash was about the same in 19.1% of rice solid soup immersion spheres and 19.2% of adzuki bean liquid soup immersion spheres.

The sensory evaluation of the distilled liquor made by distilling the soju mash in the solid soup and the red bean liquid soup was carried out by distillation under reduced pressure distillation. As shown in the figure, there is no significant difference between the solid soup and the red bean liquid soup, and it can be seen that even adzuki bean soup can be manufactured with sufficient quality. Moreover, the adzuki bean liquid soup was also commented as "plump tastes and sweet scent," suggesting the possibility of making soju circumference which is clearly different from the conventional solid soup method.

Example 19 Preparation of Sweet Potato Liquid Soup

1. Pre-culture

8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, the preculture medium was inoculated with final spores of Bacillus agar (Aspergillus kawachii IFO4308) to 1 × 10 ⁶ cells / ml, and cultured at 37 ° C. for 24 hours at 100 rpm.

2. Main culture method

Lightly wash the outside of one raw sweet potato (approximately 20 g), and process 1.0 g of potassium nitrate and 1.5 g of potassium dihydrogen phosphate as it is without processing such as cutting or peeling off the stem. A 2000 ml baffle conical flask was filled with the ml and autoclaved sterilized at 121 ° C. for 15 minutes. 1 ml of the preculture was inoculated with this main culture medium and gently shaken at 37 ° C. for 48 hours at about 80 rpm to incubate the sweet potatoes in a culture medium.

Glucoamylase (GA) activity and acid resistant α-amylase (ASAA) activity in the culture supernatant after culture were measured by the same method as in Example 15.

3. Results

It is as shown in Table 38. As mentioned above, the target value of the enzyme activity required for the preparation of soju is 100 U / ml for glucoamylase activity and 10 U / ml for acid resistant α-amylase activity.

As is apparent from Table 38, both GA and ASAA both enzymes were produced simultaneously. The ASAA did not achieve the target enzyme activity, but it could be expected to increase the enzyme activity by further optimizing liquid station culture conditions such as aeration conditions, or increase the proportion of soup in soju soaking even in the current state of sweet potato liquid soup. Soju can be manufactured sufficiently.

Example 20 Preparation of Amaranthus Liquid State

1. Preculture Method; 8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, the preculture medium was inoculated with Bacillus agar (Aspergillus kawachii IFO4308) to 1 × 10 ⁶ cells / ml, and cultured at 37 ° C. for 24 hours at 100 rpm.

2. The present culture method; An amaranthus 1-8 g, potassium nitrate 0.2 g, potassium dihydrogen phosphate 0.3 g and 100 ml of water were charged into a 500 ml baffle conical flask and autoclave sterilized for 15 minutes at 121 ° C. 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm. Glucoamylase activity (GA) and acid resistant α-amylase activity (ASAA) in the culture supernatant after culture were measured. The measurement of GA activity is performed using a glycosylation fractionation quantitative kit (manufactured by Gikkoman), and the measurement of ASAA activity is slightly improved by the method described in <Non-Patent Document 4>, and the acid-treated culture of the non-heat-resistant α- After inactivation of amylase activity, it was carried out using the α-amylase measurement kit (Gikkomann). More specifically, 9 ml of 100 mM acetic acid buffer solution (pH3) was added to 1 ml of the culture broth, and acid treatment was performed at 37 ° C for 1 hour, followed by measurement using the α-amylase activity measurement kit (Gikkomann). .

3. results; As shown in Table 39. The targets of enzyme activity necessary for brewing soju in the review so far are 100 U / ml of glucoamylase and 10 U / ml of acid resistant α-amylase. As apparent from the table, it was shown that with the amount of Amaranthus used at 2% or more, the activities of both GA and ASAA enzymes met the target values, and Amaranthus was effective as a raw material for the liquid station.

Example 21 Preparation of Quinoa Liquid Station

1. Preculture Method; 8 g of 90% white rice and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized at 121 ° C for 15 minutes. After cooling, the preculture medium was inoculated with Bacillus agar (Aspergillus kawachii IFO4308) to 1 × 10 ⁶ cells / ml, and cultured at 37 ° C. for 24 hours at 100 rpm.

2. The present culture method; 1-8 g of quinoa, 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were charged into a 500 ml baffle conical flask, and autoclave sterilized for 15 minutes at 121 ° C. 1 ml of the preculture was inoculated with this main culture medium, followed by shaking culture at 37 ° C for 48 hours at 100 rpm. Glucoamylase activity (GA) and acid resistant α-amylase activity (ASAA) in the culture supernatant after culture were measured in the same manner as in Example 20.

3. results; As shown in Table 40. The targets of enzyme activity necessary for brewing soju in the review so far are 100 U / ml of glucoamylase and 10 U / ml of acid resistant α-amylase. As is clear from the table, it was found that both enzymes were produced in a good balance with 2 to 4% of quinoa used, and that quinoa was effective as a raw material for liquid station.

According to the present invention, grains of which the surface is covered with grains or cereals (only chaff, etc.) removed only, grain-covered beans or potatoes, or amaranthus and / or quinoa of cereal grains are used as a culture material. A method for producing this stable liquid station efficiently and inexpensively is provided. This liquid station is not only suitable for the production of fermented foods and beverages, but also for the production of alcoholic beverages such as soju, since both enzymes of glucoamylase and acid resistant α-amylase are produced in a balanced and high production.

Claims (17)

CLAIMS 1. A method for producing a liquid station for use in producing fermented food and drink, the method comprising: cereal grains covered with a surface of grains as a culture raw material; Cereals with only chaff removed from the surface; And culturing the bacterium in a liquid medium comprising any one of the raw potatoes covered with an outer skin. The Bacillus is Baekkukyun or Black Baekkuk, When the liquid medium is prepared, the mixing ratio of the culture raw material to water is 1-20% (w / vol) when cereals are used as a culture raw material, and 1-10% (w) when potatoes are used as a culture raw material. / vol) A method for producing a liquid station, characterized in that the glucoamylase and acid-resistant α-amylase are simultaneously produced and accumulated in a domestic culture cultured in a liquid medium containing the culture material. delete The method for producing a liquid station according to claim 1, wherein the grain covered with the surface of the grain is 92% or more of fine white or fine white. The method according to claim 1 or 3, wherein the cereal grains are covered barley. delete 4. The process of claim 1 or 3 wherein the cereals are rice, wheat, buckwheat, blood, crude, millet, high quantity or corn. The method for producing a liquid station according to claim 1, wherein the grains obtained by removing only chaff on the surface are brown rice. The method for producing a liquid station according to claim 1, wherein the raw potatoes covered with the outer skin are sweet potatoes. delete delete A method for producing a fermented food or drink, wherein the fermented food or drink is produced using the liquid station obtained by the method of claim 1. The method for producing a fermented food or drink according to claim 11, wherein the production of the fermented food or drink is performed in a liquid phase. The method for producing a fermented food or drink according to claim 11, wherein the production of the fermented food or drink is performed in a liquid phase in a state where an outer space and a shielding state are maintained. The method for producing a fermented food or drink according to claim 11, wherein the production of the fermented food or drink is performed by adding a raw material to the liquid station to produce a primary mash. The method for producing a fermented food or drink according to claim 11, wherein the fermented food or drink is shochu. A liquid station set for the production of fermented food and drink having at least glucoamylase activity and acid resistance α-amylase activity obtained by the method for producing a liquid station of claim 1. The method of claim 1, wherein in the production of a liquid station for culturing Korean bacteria in a liquid medium containing the culture material described above, the enzyme activity of the liquid station is reduced by suppressing the release rate of sugars derived from starch in the culture material into the culture system. Method for producing a liquid station to be adjusted.

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