JP3718677B2 - Method for producing liquid cake - Google Patents

Description

  The present invention relates to a method for producing a liquid koji used for the production of fermented foods and drinks, particularly a liquid koji having an enzyme activity required for shochu brewing.

  The koji used in the production of alcoholic beverages, etc., is prepared by inoculating and cultivating spores of filamentous fungi on the raw material after processing such as cooking, and preparing a liquid medium by adding the raw material and other nutrients to water, There is a liquid koji that inoculates and cultivates gonococcal spores or precultured hyphae.

  In the production of conventional alcoholic beverages or fermented foods and beverages, such as sake, shochu, soy sauce, miso, mirin, etc., so-called solid koji produced by a solid culture method is widely used. This solid culture method can be performed using Aspergillus kawachii, Aspergillus awamori, Aspergillus niger, Aspergillus oryzae, or Aspergillus soja. This is a culturing method in which spores are sprayed on a solid raw material such as steamed cereal and the koji mold is grown on the surface.

  For example, in the production of shochu, solid soot such as Aspergillus kawachii and Aspergillus awamori is widely used. However, since the solid culture method is a culture system in which raw materials and koji molds are dispersed non-uniformly, it is difficult to make uniform factors such as temperature, water content, and various nutrient components, and the culture control is very complicated. . In addition, iron is often produced in an open state, and in this case, attention is required in terms of quality control such as contamination by various bacteria. Therefore, it can be said that the method is not suitable for large-scale manufacturing.

  In contrast, the liquid culture method is easy to control and quality control, and is a culture form suitable for efficient production. For example, the enzyme activity required for shochu brewing cannot be obtained sufficiently. There are few examples where the culture obtained by liquid culture of Aspergillus oryzae is actually used as shochu. Here, the culture obtained by the liquid culture method refers to a culture itself obtained by the liquid culture method (hereinafter also referred to as a liquid koji), a culture solution, a microbial cell, a concentrate thereof, or a dry product thereof. There may be.

  The main reason why the culture obtained by the liquid culture method is not used for the production of fermented foods and drinks such as shochu is not only the enzyme production behavior of amylase and cellulase of gonorrhea is significantly different from that of solid culture, but overall production It is known that the property decreases (see Non-Patent Document 1).

  Usually, in the production of alcoholic beverages including shochu, alcohol is produced by parallel double fermentation. Therefore, the saccharide-degrading-related enzymes of koji mold that affect glucose supply to koji molds, in particular glucoamylase and acid-resistant α-amylase, are key enzymes in alcohol fermentation. However, it is known that in the culture obtained by the liquid culture method, the activity of glucoamylase is remarkably low, and the production behavior is greatly different from that of solid culture (see Non-Patent Document 2).

  As a method for improving the glucoamylase activity of Aspergillus oryzae, a method of cultivating Aspergillus oryzae while stressing the growth of mycelia (see Patent Document 1) or a method of adding roasted cereals to an Aspergillus culture medium (see Patent Document 2) It has been reported. The method disclosed in Patent Document 1 is a method in which a novel gene glaB encoding glucoamylase is expressed by culturing in a entrapping immobilization agent on a porous membrane or having voids, thereby enhancing the enzyme activity. Special culture equipment is required and is not practical. Moreover, the method disclosed in Patent Document 2 is a method for culturing koji molds in a liquid medium using cereals roasted as at least a part of the raw material, and adds a new manufacturing process of roasting cereals. .

  Therefore, the present inventors have proposed an invention relating to a method for culturing koji molds using a liquid medium containing a saccharide that is hardly degradable for koji molds (see Patent Document 3). According to the present invention, in a liquid culture of Aspergillus oryzae, it is possible to easily and inexpensively obtain a Neisseria gonorrhoeae culture having high activity of a saccharide degradation-related enzyme such as glucoamylase, which can be used for production of alcoholic beverages or fermented foods and drinks. it can.

  On the other hand, regarding acid-resistant α-amylase, molecular biological analysis has recently started in detail (see Non-Patent Document 3). According to it, white mold has two types of amylase genes with different properties, non-acid-resistant α-amylase and acid-resistant α-amylase, but their expression patterns are greatly different. Although acid-resistant α-amylase is sufficiently produced, it is reported that acid-resistant α-amylase, which is a key enzyme for shochu brewing, is hardly produced.

  In the production of shochu, brewing is performed in a low pH environment in order to prevent the shochu moromi from becoming rotted. However, non-acid resistant α-amylase is rapidly inactivated under low pH conditions, and therefore hardly contributes to the decomposition of carbohydrates in shochu brewing. For this reason, it is indispensable for producing shochu to produce a large amount of acid-resistant α-amylase, which is thought to contribute to the saccharide degradation of shochu brewing, by liquid culture of koji mold.

In the past, although there has been a report examining the production behavior of acid-resistant α-amylase in liquid culture of Aspergillus oryzae, the method uses a synthetic medium containing peptone or citrate buffer, and the culture time is 100 hours or more. It is difficult to say that this is a method for producing liquid koji that can be applied to actual shochu brewing (see Non-Patent Document 4).
JP-A-11-225746 JP 2001-321154 A JP 2003-265165 A Iwashita K. et al: Biosci. Biotechnol. Biochem., 62, 1938-1946 (1998), Yuichi Yamane et al .: Journal of the Japan Brewing Association., 99, 84-92 (2004) 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) Nagamine K. et al: Biosci. Biotechnol. Biochem., 67, 2194-2202 (2003) Sudo S. et al: J. Ferment. Bioeng., 76,105-110 (1993), Sudo S. et al: J. Ferment. Bioeng., 77, 483-489 (1994), Shigetoshi Sudo et al .: Journal of the Japan Brewing Association ., 89, 768-774 (1994)

  However, in the method of Patent Document 3, a koji mold culture having high glucoamylase activity is a koji mold cultured in a liquid medium prepared by adding a hardly degradable carbohydrate, and is usually prepared from a culture raw material such as cereals. The liquid culture medium is not used.

  Further, although a technique for culturing koji mold in a liquid medium to obtain a koji mold culture having a high glucoamylase activity has been disclosed, a liquid koji having a high activity of acid-resistant α-amylase, which is another key enzyme in alcohol fermentation, is disclosed. None of the techniques disclosed by culturing koji molds in a liquid medium are disclosed. This acid-resistant α-amylase is generally said to be an enzyme that is not produced in liquid culture, and a liquid koji with high acid-resistant α-amylase activity has not been developed so far.

  The object of the present invention is to use liquid koji used in the production of fermented foods and drinks, in particular glucoamylase, which is a key enzyme in alcohol fermentation of shochu brewing, and liquid koji with high activity of acid-resistant α-amylase with special carbohydrates, etc. It is to provide a method for producing liquid koji by culturing koji molds in a liquid medium using raw materials of unmilled cereals instead of a special liquid medium such as adding or using roasted raw materials.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have cultured gonococci in a liquid medium using cereals whose surfaces are covered with husks, thereby producing glucoamylase and acid-resistant α-amylase. It was found that a liquid koji with enhanced enzyme activity was produced, and the present invention was completed.

  That is, the present invention provides the following.

(1) A method for producing liquid koji used for the production of fermented foods and drinks, comprising culturing white koji molds and / or black koji molds in a liquid medium containing cereals whose surfaces are covered with husks as a culture raw material . A method for producing a liquid koji for producing fermented foods and drinks , characterized in that glucoamylase and acid-resistant α-amylase are simultaneously produced and accumulated therein .

  (2) The method for producing a liquid koji according to (1), wherein the cereal is unmilled or is at least a milling ratio that is milled to the extent that the husk is left on the surface of the grain.

( 3 ) The method for producing a liquid koji according to claim 1 or 2, wherein the liquid medium contains 1 to 20% (w / vol) of cereal with respect to water.

( 4 ) The manufacturing method of fermented food / beverage products which manufactures fermented food / beverage products using the said liquid koji obtained by the method of any one of (1) to ( 3 ).

( 5 ) The method for producing a fermented food or drink according to ( 4 ), wherein all the steps are performed in a liquid phase.

( 6 ) The method for producing a fermented food / beverage product according to (4) or (5) , wherein the fermented food / beverage product is produced in a liquid phase in a state where the outside and the shielded state are maintained.

( 7 ) The method for producing a fermented food or drink according to any one of (4) to (6) , wherein the fermented food or drink is produced by placing the raw material on the liquid koji and producing a primary mash.

( 8 ) The method for producing a fermented food or drink according to any one of (4) to (7) , wherein the fermented food or drink is shochu.

( 9 ) A liquid koji set produced by the method according to any one of (1) to (3) for producing a fermented food or drink having at least glucoamylase and acid-resistant α- amylase .

  According to the present invention, a liquid in which a group of enzymes necessary for shochu brewing such as glucoamylase and acid-resistant α-amylase is simultaneously produced at high yield by culturing koji mold in a liquid medium containing cereals whose surface is covered with husk. Can produce firewood. Since liquid culture allows stricter culture control than solid culture, a liquid koji with stable quality can be produced at low cost.

  In addition, when the liquid koji produced by the present invention is used, fermentability comparable to that of shochu mash using conventional solid koji is obtained, and the produced shochu is about the same as the koji made using the solid koji. A shochu liquor that has quality and is sensuously inferior can be produced.

  Moreover, since the cereals used in the present invention are not whitened or have been at least polished to the extent that the husk is left on the surface of the grain, an improvement in raw material utilization rate and yield can be expected.

  In addition, when producing shochu using the liquid soot produced according to the present invention, unlike the conventional shochu making using solid soot, the entire process can be performed in the liquid phase, so that the efficiency is higher than in the past. And a stable shochu production system can be provided.

  Hereinafter, the present invention will be specifically described.

In the method for producing liquid koji according to the present invention, koji molds are cultured in a liquid medium prepared by adding raw materials such as cereals to produce a liquid koji having enhanced enzyme activities of glucoamylase and acid-resistant α-amylase. The process is included. That is, since the koji mold is cultured using the above-mentioned cereal as a raw material, it takes time for saccharification of starch in the cereal, the sugar release rate to the culture system is suppressed, and the enzyme activity of the liquid koji is enhanced. . Moreover, glucoamylase and acid-resistant α-amylase are simultaneously generated and accumulated in a balanced manner.
In the present invention, examples of cereals used as a raw material include barley, rice, wheat, buckwheat, millet, millet, millet, cucumber, and corn. As the shape of these raw materials, it is possible to use an unmilled product, or at least a milling ratio that has been milled to such an extent that the husk remains on the surface of the grain. For example, when the cereal is barley, the unpolished milling rate is 100%, or the unpolished milling rate is 100%, and the unpolished milling rate (100%) is used to obtain the barley grain ratio (general Is a ratio obtained by subtracting 7 to 8%), that is, a fineness ratio of about 92 to 93% or more.

  Here, the milling ratio refers to the ratio of cereals left after cerealing, and for example, the milling ratio of 90% means that 10% of the skin of the surface layer of the cereal is scraped off. Moreover, in the present invention, the unpolished barley includes unpolished wheat to those that have been polished to such an extent that the husk remains on the surface of the grain, that is, those having a polishing ratio of 90% or more. Moreover, a grain skin means the outer side part which has covered the surface of the grain of cereals.

Raw grain is mixed with water to prepare a liquid medium. The blending ratio of this cereal is adjusted so that glucoamylase and acid-resistant α-amylase are selectively generated and accumulated during the cultivation of Aspergillus. For example, when barley is used as a raw material, it is prepared in a liquid medium in which 1 to 20% (w / vol) of brown barley is added to water. In addition, when unpolished barley is used as brown wheat, it is more preferably prepared in a liquid medium supplemented with 8 to 10% (w / vol). More preferably, it is prepared in a liquid medium supplemented with 1 to 4% (w / vol).
As described above, the optimum blending amount varies depending on the degree of whitening of the raw material used, the koji strain used, the type of raw material, and the like, and may be arbitrarily selected.

  When gonococcus is cultured in a liquid medium supplemented with 1 to 20% (w / vol) of barley, glucoamylase and acid-resistant α-amylase enzymes are produced in a well-balanced manner, and in particular, 8 to 10% (w / vol) of barley is produced. ) With the added liquid medium, a liquid koji having sufficient enzyme activity for use in shochu brewing can be obtained. When the amount of brown barley used exceeds 10% (w / vol), the viscosity of the culture solution becomes high, the supply of oxygen and air necessary for aerobic culture of koji mold is insufficient, and the oxygen concentration in the culture Is not preferable because the culture is difficult to proceed.

  The starch contained in the raw material may be gelatinized before culturing. The starch gelatinization method is not particularly limited, and may be performed according to a conventional method such as a steaming method or a roasting method. In the sterilization step of the liquid medium described later, when the starch is heated to a starch gelatinization temperature or higher by high-temperature high-pressure sterilization or the like, the starch gelatinization is simultaneously performed by this treatment.

  In addition to the aforementioned raw materials, it is preferable to add organic substances, inorganic salts, and the like as nutrient sources to the liquid medium. These additives are not particularly limited as long as they are generally used for culturing koji mold, but organic substances include rice bran, wheat straw, corn steep liquor, soybean meal, defatted soybean, etc., and inorganic salts are ammonium salts. , Nitrates, potassium salts, acidic phosphates, calcium salts, magnesium salts, and the like, and two or more organic substances and / or inorganic salts may be used at the same time. These addition amounts are not particularly limited as long as they promote the growth of Neisseria gonorrhoeae, but are about 0.1 to 5% (w / vol) as organic substances and 0.1 to 1% (w / vol.) As inorganic salts. It is preferable to add about vol. The liquid medium of Aspergillus thus obtained may be sterilized as necessary, and the treatment method is not particularly limited. As an example, a high-temperature and high-pressure sterilization method can be mentioned, which may be performed at 121 ° C. for 15 minutes.

The sterilized liquid medium is cooled to the culture temperature, and then the koji mold is inoculated into the liquid medium. The koji mold used in the present invention is a koji mold having an ability to produce a saccharide-degrading enzyme, preferably a koji mold having a glucoamylase producing ability and an acid-resistant α-amylase producing ability, and is represented by, for example, Aspergillus kawachii. that white koji, black koji mold and the like typified by Aspergillus awamori (Aspergillus awamori) and Aspergillus niger (Aspergillus niger) or the like. Moreover, the form of the koji mold inoculated into the medium is arbitrary, and spores or hyphae can be used.

These koji molds can be used either by culturing with one kind of strain or mixed culturing with two or more kinds of the same or different kinds of strains. There is no problem whether these are used in the form of spores or hyphae obtained by preculture, but it is preferable to use hyphae because the time required for the logarithmic growth phase is shortened. There is no particular limitation on the amount of koji mold inoculated into the liquid medium, but about 1 × 10 ⁴ to 1 × 10 ⁶ spores per 1 ml of the liquid medium, and 0.1 to 10 of the preculture solution for mycelia. It is preferable to inoculate about 1%.

  The culture temperature of the koji mold is not particularly limited as long as it does not affect the growth, but it is preferably 25 to 45 ° C, more preferably 30 to 40 ° C. If the culture temperature is low, the growth of Aspergillus oryzae slows down, and contamination with various bacteria tends to occur. The culture time is preferably 24 to 72 hours. Any culture apparatus may be used as long as it can perform liquid culture. However, since Neisseria gonorrhoeae needs to perform aerobic culture, it needs to be performed under aerobic conditions in which oxygen and air can be supplied into the medium. Moreover, it is preferable to stir so that the raw material, oxygen, and koji mold in the medium are uniformly distributed in the apparatus during the culture. The stirring conditions and the aeration amount may be any conditions as long as the culture environment can be maintained aerobically, and may be appropriately selected depending on the culture apparatus, the viscosity of the medium, and the like.

  By culturing by the above culture method, glucoamylase and acid-resistant α-amylase enzymes are simultaneously produced in a well-balanced manner, and a liquid koji having enzyme activity that can be used for shochu brewing is obtained. In addition, the liquid koji obtained by the above culture method may be a culture solution obtained by centrifuging the culture, a concentrate thereof, a dry product thereof, or the like in addition to the culture itself.

  The liquid koji obtained by the production method of the present invention can be used for the production of alcoholic beverages or fermented foods and drinks. For example, when producing sake, when producing shochu at the mash mother and each mash preparation stage, when producing soy sauce at the mash preparation stage, when producing miso at the preparation stage In the preparation stage, when producing mirin, liquid soot can be used instead of solid soot in the preparation stage.

  Moreover, when manufacturing liquor or fermented food / beverage products using the culture liquid obtained from the above-mentioned liquid koji or culture, or those concentrates, all processes can be performed in a liquid phase. As a method for producing alcoholic beverages in which all steps are performed in a liquid phase, for example, when producing shochu, corn, wheat, rice, potato, sugar cane, etc. are used as raw materials, and the raw materials are used at a high temperature of about 80 ° C. A method of producing a mash that has been subjected to alcohol fermentation by adding the above-described liquid koji and yeast after being melted and liquefied using an agent, by an atmospheric distillation method or a vacuum distillation method, etc. It is done.

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

<Experimental example 1> [Examination of the amount of brown wheat used in the production of liquid koji]
Five types of liquid media were prepared by changing the ratio of raw barley as shown in Table 1, and koji molds were cultured in the respective liquid media to produce liquid koji.

  First, brown wheat becomes 1, 2, 4, 8, 10% (w / vol) in water added with potassium nitrate 0.2% (w / vol) and potassium dihydrogen phosphate 0.3% (w / vol) Five types of liquid media added as described above were prepared. For each liquid medium, 100 ml of the prepared liquid medium was placed in a 500 ml baffled Erlenmeyer flask, sterilized by autoclave, and pre-cultured in white liquid (Aspergillus kawachii IFO4308) in 1% (v / Vol). In addition, the unpolished white wheat barley used for domestic barley was used.

  Thereafter, the cells were cultured for 48 hours at a temperature of 37 ° C. and a shaking speed of 100 rpm. After completion of the culture, the amount of glucoamylase and acid-resistant α-amylase produced was measured for each of the obtained cultures. Table 1 and FIG. 1 show the amounts of glucoamylase and acid-resistant α-amylase produced in the culture obtained by culturing in the liquid medium according to the amount of brown wheat used. In addition, the saccharification power fractionation determination kit (made by Kikkoman) was used for the measurement of the enzyme activity of glucoamylase. In addition, the enzyme activity of acid-resistant α-amylase was measured by slightly improving the method described in <Non-Patent Document 3> and inactivating the non-acid-resistant α-amylase by acid treatment of the culture. Acid-resistant α-amylase was measured using an α-amylase measurement kit (manufactured by Kikkoman). More specifically, 9 ml of 100 mM acetate buffer (pH 3) was added to 1 ml of the culture solution, and after acid treatment at 37 ° C. for 1 hour, measurement was performed using an α-amylase measurement kit (manufactured by Kikkoman).

  On the other hand, a liquid medium was prepared as in the test section using the wheat that had been refined with 70% white milling ratio of domestic two-row barley as the raw material, and prepared in the same manner as in the test section. After culturing and after completion of the culture, the amounts of glucoamylase and acid-resistant α-amylase produced were measured for each of the obtained cultures. Table 1 and FIG. 2 show the amounts of glucoamylase and acid-resistant α-amylase produced in the cultures obtained by culturing in the liquid medium according to the amount of barley used.

As shown in Table 1 and FIG. 1, cultivated using brown barley produced a high balance of glucoamylase and acid-resistant α-amylase simultaneously with the increase in the amount of brown wheat used, and the amount produced was also high. It was confirmed that there was a significant increase compared to using control barley. In particular, in a liquid medium supplemented with 10% (w / vol) brown barley, 217.3 U / ml of glucoamylase and 14.0 U / ml of acid-resistant α-amylase were produced, which is sufficient for use in shochu brewing. Enzyme activity was obtained at the same time. (For reference, the enzyme activity values of glucoamylase and acid-resistant α-amylase necessary for shochu production are 100 U / ml or more for glucoamylase and 10 U / ml or more for acid-resistant α-amylase.)
On the other hand, in the control group using round barley, as shown in Table 1 and FIG. 2, the glucoamylase activity is maximum in a liquid medium supplemented with 2% (w / vol) of barley, and the acid-resistant α-amylase activity is round barley. In the liquid medium supplemented with 8% (w / vol), both enzymes were not produced at the same time.

  Thus, glucoamylase and acid-resistant α-amylase are simultaneously produced in a well-balanced manner by culturing koji molds in a liquid medium supplemented with 1 to 20% (w / vol) of brown wheat. In particular, the addition of 8 to 10% (w / vol) of brown barley (unrefined white) made it possible to produce a liquid koji that can simultaneously provide sufficient enzyme activity for use in shochu brewing.

  When gonococcus is cultured in a liquid medium using brown barley, glucoamylase and acid-resistant α-amylase are simultaneously produced in a well-balanced and high quality by using brown wheat whose surface is covered with husk as a raw material. The release of sugars such as glucose derived from the starch in the starch is suppressed by the husk, and the sugar concentration in the culture is relatively low, so that enzymes such as glucoamylase and acid-resistant α-amylase are produced. This is thought to be due to the ease.

<Example 1> [Production of liquid koji using brown wheat]
First, a liquid medium added with 0.2% (w / vol) potassium nitrate and 0.3% (w / vol) potassium dihydrogen phosphate in water so that brown wheat becomes 10% (w / vol) Prepared. Next, 100 ml of the prepared liquid medium was put into a 500 ml baffled Erlenmeyer flask, sterilized by autoclave, and pre-cultured with Aspergillus kawachii IFO4308 in 1% (v / vol) with respect to the liquid medium. Inoculated to become. In addition, the unpolished white wheat barley used for domestic barley was used.

  Thereafter, the cells were cultured for 48 hours at a temperature of 37 ° C. and a shaking speed of 100 rpm. After completion of the culture, the amount of glucoamylase and acid-resistant α-amylase produced was measured for each of the obtained cultures. As a result, glucoamylase was 217.3 U / ml and acid-resistant α-amylase was 14.0 U / ml. Enzyme activity sufficient for use in shochu brewing was obtained at the same time.

<Example 2> [Production of barley shochu using liquid rice bran]
Production of shochu using liquid koji (culture product with enhanced glucoamylase and acid-resistant α-amylase) obtained by culturing in a liquid medium prepared by adding 10% (w / vol) of barley in Example 1. Went.

  That is, using 500 ml of a liquid koji obtained by culturing in a liquid medium prepared by adding 10% (w / vol) of the unpolished barley in Example 1, with the preparation composition shown in Table 2, total wheat 1328.6 g was charged, the fermentation temperature was kept at 25 ° C., and three-stage charging was performed for 5 days for primary charging, 2 days for secondary charging, and 13 days for tertiary charging. As the wheat, 70% white milled domestic 2 barley was washed with water, soaked for 60 minutes, drained for 30 minutes, and then steamed for 35 minutes. In addition, in the primary charging, the amount of 50.0 g of wheat brought from the liquid koji was insufficient for fermentation, so 262.9 g of wheat was charged so that the same amount of wheat as the solid koji was charged. In addition, shochu yeast (Kagoshima yeast) was used for yeast, and 50 microliters of what was statically cultured at 30 degreeC for 48 hours by YPD culture medium was inoculated.

  In addition, as a control preparation (solid rice cake preparation), shochu production was carried out using the solid soba noodles with the preparation composition shown in Table 3. The method for producing solid koji is 70% polished wheat, washed 40 minutes, soaked for 40 minutes, drained for 30 minutes, cooked for 40 minutes, allowed to cool to 40 ° C., and allowed to cool to 40 ° C. kawachii IFO4308) was inoculated and cultured for 24 hours at 40 ° C. and 95% relative humidity, for 6 hours at 35 ° C. and 95% relative humidity, and for 18 hours at 30 ° C. and 90% relative humidity. The fermentation conditions and the like were the same as the above-described preparation of the present invention (liquid koji preparation).

  The fermentation process is shown in FIG. 3 in contrast to the control solid koji preparation. As is clear from FIG. 3, the fermentation process using the liquid koji showed almost the same fermentation process as compared to the control kneading using the solid koji. In addition, the alcohol content of the obtained final mash was 17.8%, which was the same for both the liquid mash and the solid mash.

  Next, the raw sake obtained by distilling the final mash obtained under reduced pressure was hydrated to 25% alcohol, and then sensory by a scoring method (5-point evaluation method, 1: good to 5: bad) by 8 panelists. Evaluation was performed and the average score is shown in Table 4.

  As a result, it was confirmed that there was almost no difference in liquor quality, and it was possible to produce shochu with the same liquor quality as when using solid koji, even when using liquid koji.

  From the above results, according to the present invention, glucoamylase and acid-resistant α-amylase are simultaneously produced in a well-balanced manner by culturing koji molds in a liquid medium supplemented with 1-20% (w / vol) brown wheat. In particular, the addition of 8 to 10% (w / vol) of unpolished barley (unrefined white) made it possible to produce a liquid koji that can simultaneously provide sufficient enzyme activity for use in shochu brewing. For this reason, by using this liquid lees, it has become possible to produce a liquor with a liquor quality equivalent to that produced using solid lees. Furthermore, a liquid koji with high glucoamylase activity and acid-resistant α-amylase activity can be produced in a simple liquid medium without strict culture control using special culture equipment or special culture engineering techniques. Moreover, stable production of high-quality sputum has become possible by carrying out extremely strict control of sputum production compared to solid culture. Furthermore, the liquefaction of koji enables not only simplification of fermentation management by fluidizing moromi, but also labor saving and efficiency of the koji manufacturing process and eventually the shochu manufacturing process.

<Example 3> [Production of rice shochu by liquid rice bran using buckwheat]
1. Solid rice bran production method 90% polished rice, washed for 15 minutes, drained for 10 minutes, drained for 10 minutes, boiled for 30 minutes, allowed to cool to 40 ° C, 1 g of seed meal per 1 kg of polished rice (white birch Aspergillus kawachii IFO4308)) And cultured at 40 ° C. and 95% relative humidity for 24 hours, at 35 ° C. and 95% relative humidity for 6 hours, and at 30 ° C. and 90% relative humidity for 18 hours.

2. Method for producing liquid koji (1) Pre-culture method: 8 g of 90% polished rice and 100 ml of water were placed in a 500 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. After standing to cool, seed culture spores of Aspergillus kawachii IFO4308 were inoculated at 1 × 10 ⁶ cells / ml in the preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main 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 put into a 2000 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. A soba liquid koji was produced by inoculating 5 ml of the preculture solution into this main culture medium, followed by shaking culture at 37 ° C. for 48 hours at 100 rpm. The enzyme activity of the liquid koji at this time was GA activity of 112.4 U / ml and ASAA activity of 10.4 U / ml.

3. Rice shochu production method (1) Yeast used; Shochu yeast (Kagoshima yeast)
(2) Preparation formulation; Table 5 and Table 6 show the preparation formulation. The rice used was 90% polished rice, which was soaked for 15 minutes, drained for 10 minutes, and steamed for 30 minutes. There were two test sections, 1) solid dredging and 2) buckwheat liquid dredging, and the total rice and pumped water in both test plots were blended in the same amount. Yeast was inoculated in a YPD medium at 30 ° C. for 48 hours and inoculated with 50 μl.
(3) Fermentation conditions; constant at 25 ° C (4) Distillation conditions;

  The fermentation process is shown in FIG. 4 in comparison with the control solid koji preparation. As is clear from FIG. 4, the fermentation process using the buckwheat liquid koji showed almost the same fermentation process as compared to the control kneading using the solid koji. In addition, the alcohol content of the final moromi in the obtained solid koji charging zone and buckwheat liquid koji charging zone was approximately the same as 19.1% and 18.9%, respectively.

  Next, the sensory evaluation of the shochu liquor produced by distilling the shochu moromi of the obtained solid koji and soba liquid koji prepared by a vacuum distillation method was conducted using a five-point evaluation method for six specialized panels (Ryo 1-3). No. 5), it was found that there was no big difference between the solid koji and soba liquid koji preparation areas, and it was possible to produce a shochu with sufficient quality even with soba liquid koji. Furthermore, as shown in Table 7, in the soba liquid koji district, there was also a comment that “soba flavored”, and it was confirmed that the use of soba liquid koji can add a “soba” character to rice shochu.

<Example 4> [Production of rice shochu by liquid koji using millet]
1. Solid rice bran production method 90% polished rice, washed for 15 minutes, drained for 10 minutes, drained for 10 minutes, boiled for 30 minutes, allowed to cool to 40 ° C, 1 g of seed meal per 1 kg of polished rice (white birch Aspergillus kawachii IFO4308)) And cultured at 40 ° C. and 95% relative humidity for 24 hours, at 35 ° C. and 95% relative humidity for 6 hours, and at 30 ° C. and 90% relative humidity for 18 hours.

2. Method for producing liquid koji (1) Preculture method: 8 g of 90% polished rice and 100 ml of water were placed in a 500 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. After standing to cool, seed culture spores of Aspergillus kawachii IFO4308 were inoculated at 1 × 10 ⁶ cells / ml in the preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main 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 put in a 2000 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. The main culture medium was inoculated with 5 ml of the preculture solution, and cultured with shaking at 37 ° C. for 48 hours at 100 rpm to produce millet liquid koji. The enzyme activity of the liquid koji at this time was GA activity 101.3 U / ml and ASAA activity 11.0 U / ml.

3. Rice shochu production method (1) Yeast used; Shochu yeast (Kagoshima yeast)
(2) Preparation formulation; Table 8 and Table 9 show the preparation formulation. The rice used was 90% polished rice, which was soaked for 15 minutes, drained for 10 minutes, and steamed for 30 minutes. There were two test sections: 1) solid dredging and 2) millet liquid dredging. The total rice and pumped water in both test plots were blended in the same amount. Yeast was inoculated in a YPD medium at 30 ° C. for 48 hours and inoculated with 50 μl.
(3) Fermentation conditions; constant at 25 ° C (4) Distillation conditions;

  The fermentation process is shown in FIG. 5 in comparison with the control of solid koji. As is clear from FIG. 5, the fermentation process using millet liquid koji showed almost the same fermentation process as compared to the control kneading using solid koji. In addition, the alcohol content of the final moromi in the obtained solid koji charging zone and millet liquid koji charging zone was 19.1%, which was the same.

  Next, sensory evaluation of shochu raw sake produced by distilling shochu moromi from the solid koji and millet liquid koji prepared by the vacuum distillation method, a five-point evaluation method for six specialist panels (good 1-3-5 bad) As a result, it was found that there was no significant difference between the solid soot preparation section and the millet liquid soot preparation section, and it was possible to produce a shochu with sufficient quality even with the millet liquid soot. Furthermore, as shown in Table 10, there was also a comment that the Awa liquid koji district had a “fresh taste”, suggesting the possibility of producing rice shochu raw sake with a liquor quality different from the conventional solid koji method. .

<Example 5> [Production of rice shochu using liquid rice bran]
1. Solid rice bran production method 90% polished rice, washed for 15 minutes, drained for 10 minutes, drained for 10 minutes, boiled for 30 minutes, allowed to cool to 40 ° C, 1 g of seed meal per 1 kg of polished rice (white birch Aspergillus kawachii IFO4308)) And cultured at 40 ° C. and 95% relative humidity for 24 hours, at 35 ° C. and 95% relative humidity for 6 hours, and at 30 ° C. and 90% relative humidity for 18 hours.

2. Method for producing liquid koji (1) Preculture method: 8 g of 90% polished rice and 100 ml of water were placed in a 500 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. After standing to cool, seed culture spores of Aspergillus kawachii IFO4308 were inoculated at 1 × 10 ⁶ cells / ml in the preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main culturing method: Japanese cypress 40 g, potassium nitrate 1.0 g, potassium dihydrogen phosphate 1.5 g and water 500 ml were placed in a 2000 ml baffled conical flask and autoclaved at 121 ° C. for 15 minutes. The main culture medium was inoculated with 5 ml of the preculture solution, and cultured with shaking at 37 ° C. for 48 hours at 100 rpm, to produce a fly liquid koji. The enzyme activity of the liquid koji at this time was GA activity 113.0 U / ml and ASAA activity 10.2 U / ml.

3. Rice shochu production method (1) Yeast used; Shochu yeast (Kagoshima yeast)
(2) Preparation formulation: The preparation formulation is shown in Table 11 and Table 12. The rice used was 90% polished rice, which was soaked for 15 minutes, drained for 10 minutes, and steamed for 30 minutes. There were two test sections, 1) solid dredging and 2) millet liquid dredging, and the total amount of rice and the amount of pumped water in both test plots were blended to be the same amount. Yeast was inoculated in a YPD medium at 30 ° C. for 48 hours and inoculated with 50 μl.
(3) Fermentation conditions; constant at 25 ° C (4) Distillation conditions;

  The fermentation process is shown in FIG. 6 in contrast to the control solid koji preparation. As is clear from FIG. 6, compared with the control feed using the solid koji, the same fermentation process was shown in the feed using the millet liquid koji. Moreover, the alcohol content of the final moromi of the obtained solid koji charging zone and the millet liquid koji charging zone was approximately the same at 19.1% and 18.8%, respectively.

  Next, sensory evaluation of shochu raw sake produced by distilling shochu moromi of the solid koji and koji liquid koji prepared by the vacuum distillation method, a five-point evaluation method for six specialist panels (good 1-3-5 bad) As a result, it was found that there was no significant difference between the solid koji liquid feed area and the millet liquid koji charge area, and it was possible to produce a shochu with sufficient quality even with the fly liquid koji. Furthermore, as shown in Table 13, there was also a comment that the millet liquid cocoon ward had a “clean taste”, suggesting the possibility of producing rice shochu raw sake with a quality different from that of the conventional solid brewing method.

<Example 6> [Production of rice shochu by liquid rice cake using millet]
1. Solid rice bran production method 90% polished rice, washed for 15 minutes, drained for 10 minutes, drained for 10 minutes, boiled for 30 minutes, allowed to cool to 40 ° C, 1 g of seed meal per 1 kg of polished rice (white birch Aspergillus kawachii IFO4308)) And cultured at 40 ° C. and 95% relative humidity for 24 hours, at 35 ° C. and 95% relative humidity for 6 hours, and at 30 ° C. and 90% relative humidity for 18 hours.

2. Method for producing liquid koji (1) Preculture method: 8 g of 90% polished rice and 100 ml of water were placed in a 500 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. After standing to cool, seed culture spores of Aspergillus kawachii IFO4308 were inoculated at 1 × 10 ⁶ cells / ml in the preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main culture method; Millet 40 g, potassium nitrate 1.0 g, potassium dihydrogen phosphate 1.5 g and water 500 ml were put into a 2000 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. A millet liquid koji was produced by inoculating 5 ml of the preculture solution into this main culture medium and shaking culture at 37 ° C. for 48 hours at 100 rpm. The enzyme activity of the liquid koji at this time was GA activity 90.3 U / ml and ASAA activity 8.5 U / ml.

3. Rice shochu production method (1) Yeast used; Shochu yeast (Kagoshima yeast)
(2) Preparation formulation; Table 14 and Table 15 show the preparation formulation. The rice used was 90% polished rice, which was soaked for 15 minutes, drained for 10 minutes, and steamed for 30 minutes. There were two test sections, 1) solid dredging and 2) millet liquid dredging, and the total amount of rice and the amount of pumped water in both test plots were blended in the same amount. Yeast was inoculated in a YPD medium at 30 ° C. for 48 hours and inoculated with 50 μl.
(3) Fermentation conditions; constant at 25 ° C (4) Distillation conditions;

  The fermentation process is shown in FIG. 7 in comparison with the control solid koji preparation. As is clear from FIG. 7, the fermentation process using millet liquid koji showed almost the same fermentation process as compared to the control kneading using solid koji. Moreover, the alcohol content of the final moromi of the obtained solid koji preparation group and millet liquid koji preparation group was approximately the same at 19.1% and 18.8%, respectively.

  Next, sensory evaluation of shochu raw sake produced by distilling shochu moromi of solid koji and millet liquid koji prepared by vacuum distillation method, a five-point evaluation method for six specialized panels (good 1-3-5 bad) As a result, it was found that there was no significant difference between the solid koji and millet liquid koji preparation zones, and it was possible to produce shochu with sufficient quality even with millet liquid koji. Furthermore, as shown in Table 16, there was a comment that millet liquid cocoon ward was "rounded in taste, sweetness", suggesting the possibility of producing rice shochu raw sake with a quality different from that of the conventional solid koji method. .

<Example 7> [Manufacture of rice shochu by liquid koji using cuoliang]
1. Solid rice bran production method 90% polished rice, washed for 15 minutes, drained for 10 minutes, drained for 10 minutes, boiled for 30 minutes, allowed to cool to 40 ° C, 1 g of seed meal per 1 kg of polished rice (white birch Aspergillus kawachii IFO4308)) And cultured at 40 ° C. and 95% relative humidity for 24 hours, at 35 ° C. and 95% relative humidity for 6 hours, and at 30 ° C. and 90% relative humidity for 18 hours.

2. Method for producing liquid koji (1) Preculture method: 8 g of 90% polished rice and 100 ml of water were placed in a 500 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. After standing to cool, seed culture spores of Aspergillus kawachii IFO4308 were inoculated at 1 × 10 ⁶ cells / ml in the preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main culturing method: 40 g of Kolyang, 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate and 500 ml of water were placed in a 2000 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. A 5 ml pre-culture solution was inoculated into this main culture medium, and cultured at 37 ° C. for 48 hours with shaking at 100 rpm to produce a cucumber liquid koji. The liquid sputum enzyme activity at this time was GA activity 111.2 U / ml and ASAA activity 10.5 U / ml.

3. Rice shochu production method (1) Yeast used; Shochu yeast (Kagoshima yeast)
(2) Preparation formulation: The preparation formulation is shown in Table 17 and Table 18. The rice used was 90% polished rice, which was soaked for 15 minutes, drained for 10 minutes, and steamed for 30 minutes. The test areas were 1) charged with solid dredging, and 2) charged with cucumber liquid dredging, and the total amount of rice in both test areas and the amount of pumped water were blended in the same amount. Yeast was inoculated in a YPD medium at 30 ° C. for 48 hours and inoculated with 50 μl.
(3) Fermentation conditions; constant at 25 ° C (4) Distillation conditions;

  The fermentation process is shown in FIG. 8 in comparison with the control solid koji preparation. As is clear from FIG. 8, the fermentation process using the sorghum liquid koji showed almost the same fermentation process as compared to the control feed using the solid koji. In addition, the alcohol content of the final moromi in the obtained solid koji charging zone and the Koryang liquid koji charging zone was 19.1%, which was the same.

  Next, the sensory evaluation of shochu raw sake produced by distilling the shochu moromi of the solid koji and kourian liquid koji prepared by the vacuum distillation method is a five-point evaluation method (good 1-3-5 bad) of six specialist panels. As a result, it was found that there was no significant difference between the solid soup preparation zone and the sorghum liquid soup stock zone, and it was found that sufficient quality shochu production was possible even with the sorghum liquid soup stock. In addition, as shown in Table 19, there is a comment that “Kolyang liquid cocoon area is more cereal-like sweet”, suggesting the possibility of producing shochu liquor that is clearly differentiated from the conventional solid koji method. It was done.

<Example 8> [Manufacture of liquid koji using corn]
(1) Pre-culture method: 8 g of 90% polished rice and 100 ml of water were placed in a 500 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. After being allowed to cool, seed culture spores of Aspergillus kawachii IFO4308 were inoculated at 1 × 10 ⁶ cells / ml in this preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main 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 placed in a 500 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. 1 ml of the preculture was inoculated into this main culture medium, and cultured with shaking at 37 ° C. for 48 hours at 100 rpm. The amount of enzyme produced in the culture supernatant after culturing, ie, glucoamylase (GA) activity and acid-resistant α-amylase (ASAA) activity, was measured by the method described in Experimental Example 1. The results are shown in Table 20.

As is clear from Table 20, glucoamylase 100 U / ml, which is the target value of the enzyme activity necessary for shochu brewing, was cleared in the test plot where the amount of corn used was 4% or more. On the other hand, the target value of acid-resistant α-amylase was 10 U / ml, and although the target value was not reached, it was confirmed that the production amount of the enzyme increased as the amount of corn used increased.
As described above, although the target value of ASAA activity could not be cleared in this test, it was shown that it has the ability to produce GA enzyme and ASAA enzyme simultaneously. There is a high possibility of increasing to the value level. In addition, for example, in the preparation of shochu using 8% corn liquid koji, it was speculated that if the koji ratio was increased from the usual blend, shochu could be produced sufficiently.

<Example 9> [Production of liquid rice cake using brown wheat]
(1) Pre-culture method: 8 g of 65% white wheat and 100 ml of water were put into a 500 ml baffled Erlenmeyer flask and sterilized by autoclaving at 121 ° C. for 15 minutes. After being allowed to cool, seed spore of Aspergillus awamori IFO4388 was inoculated at 1 × 10 ⁶ cells / ml in this preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main culture method: 1-8 g of brown wheat (95% white wheat), 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate and 100 ml of water are placed in a 500 ml baffled Erlenmeyer flask and autoclaved at 121 ° C. for 15 minutes. did. 1 ml of the preculture was inoculated into this main culture medium, and cultured with shaking at 37 ° C. for 48 hours at 100 rpm. The amount of enzyme produced in the culture supernatant after culturing, ie, glucoamylase (GA) activity and acid-resistant α-amylase (ASAA) activity, was measured by the method described in Experimental Example 1. The results are shown in Table 21.

  As is clear from Table 21, in the test group with 4% of barley consumption, both glucoamylase 100 U / ml and acid-resistant α-amylase 10 U / ml, which are target values of enzyme activity necessary for shochu brewing, were cleared. From this, it was confirmed that even if black koji mold was used, the effect of high enzyme production was achieved as in the case of white koji mold.

<Example 10> [Manufacture of liquid rice bran using brown rice (rice with rice husk)]
(1) Pre-culture method: 8 g of 90% polished rice (rice rice) and 100 ml of water were put into a 500 ml baffled Erlenmeyer flask and sterilized by autoclaving at 121 ° C. for 15 minutes. After being allowed to cool, seed culture spores of Aspergillus kawachii IFO4308 were inoculated at 1 × 10 ⁶ cells / ml in this preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main culture method: 1-8 g of brown rice (with rice husk), 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate and 100 ml of water were put into a 500 ml baffled conical flask and autoclaved at 121 ° C. for 15 minutes. In addition, the brown rice used was used without threshing one with a husk (chaff). 1 ml of the preculture was inoculated into this main culture medium, and cultured with shaking at 37 ° C. for 48 hours at 100 rpm. The amount of enzyme produced in the culture supernatant after culturing, ie, glucoamylase (GA) activity and acid-resistant 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, both the glucoamylase 100 U / ml and the acid-resistant α-amylase 10 U / ml, which are target values of the enzyme activity necessary for shochu brewing, were cleared in the test plot where the brown rice consumption was 4%. From this, the enzyme high production effect was confirmed also about the rice with husk | husk (rice husk) similarly to brown wheat.

<Example 11> [Manufacture of liquid rice bran using brown rice (rice with rice husk)]
(1) Pre-culture method: 8 g of 90% polished rice (rice rice) and 100 ml of water were put into a 500 ml baffled Erlenmeyer flask and sterilized by autoclaving at 121 ° C. for 15 minutes. After being allowed to cool, seed spore of Aspergillus awamori IFO4388 was inoculated at 1 × 10 ⁶ cells / ml in this preculture medium, and cultured with shaking at 37 ° C. for 24 hours at 100 rpm.
(2) Main culture method: 1-8 g of brown rice (with rice husk), 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate and 100 ml of water were put into a 500 ml baffled conical flask and autoclaved at 121 ° C. for 15 minutes. In addition, the brown rice used was used without threshing one with a husk (chaff). 1 ml of the preculture was inoculated into this main culture medium, and cultured with shaking at 37 ° C. for 48 hours at 100 rpm. The amount of enzyme produced in the culture supernatant after culturing, ie, glucoamylase (GA) activity and acid-resistant α-amylase (ASAA) activity, was measured by the method described in Experimental Example 1. The results are shown in Table 23.

  As is clear from Table 23, both the glucoamylase 100 U / ml and the acid-resistant α-amylase 10 U / ml, which are target values of the enzyme activity necessary for shochu brewing, were cleared in the test plot where the brown rice consumption was 8%. From this, the enzyme high production effect was confirmed even when using rice with husk (rice husk) and using black koji mold.

  INDUSTRIAL APPLICABILITY According to the present invention, a method for efficiently and inexpensively producing a liquid koji having a stable quality using cereals as a culture raw material is provided. In addition, this liquid koji is suitable for the production of fermented foods and beverages, and since both glucoamylase and acid-resistant α-amylase are produced in a well-balanced manner, it is suitable for the production of alcoholic beverages such as shochu. .

It is a figure which shows the relationship between the amount of brown barley used in the Aspergillus culture using the liquid medium which uses brown wheat, and the production amount of glucoamylase and acid-resistant alpha-amylase. It is a figure which shows the relationship between the amount of barley used in the Aspergillus culture using the liquid culture medium which uses the barley of a control group, and the production amount of glucoamylase and acid-resistant alpha-amylase. It is a figure which shows the fermentation progress in the barley shochu manufacture by the liquid rice bran using brown wheat. It is a figure which shows the fermentation progress in the rice shochu manufacture by the liquid rice bran using buckwheat. It is a figure which shows the fermentation progress in the rice shochu manufacture by the liquid rice bran using millet. It is a figure which shows the fermentation progress in the rice shochu manufacture by the liquid rice bran using millet. It is a figure which shows the fermentation progress in the rice shochu manufacture by the liquid rice bran using millet. It is a figure which shows the fermentation progress in the rice shochu manufacture by the liquid rice bran using a cucumber.

Claims (9)

A method for producing liquid koji used in the production of fermented foods and drinks, comprising cultivating white koji mold and / or black koji fungus in a liquid medium containing cereals whose surface is covered with husk as a culture raw material, A method for producing a liquid koji for producing a fermented food or drink , characterized in that amylase and acid-resistant α-amylase are simultaneously produced and accumulated .   The method for producing a liquid koji according to claim 1, wherein the cereal is unmilled or more than a milling ratio that is milled to such an extent that at least the husk is left on the surface of the grain. The method for producing a liquid koji according to claim 1 or 2, wherein the liquid medium contains 1 to 20% (w / vol) of cereal with respect to water. The manufacturing method of fermented food / beverage products which manufactures fermented food / beverage products using the liquid rice cake obtained by the method of any one of Claim 1 to 3 . The manufacturing method of fermented food / beverage products of Claim 4 with which manufacture of fermented food / beverage products is performed by all the processes in a liquid phase. The manufacturing method of fermented food / beverage products of Claim 4 or 5 performed by manufacture of the fermented food / beverage products in the liquid phase of the state by which the external world and the shielding state were maintained. The method for producing a fermented food or drink according to any one of claims 4 to 6 , wherein the production of the fermented food or drink is performed by charging the liquid koji with a raw material to produce a primary mash. The method for producing a fermented food or drink according to any one of claims 4 to 7 , wherein the fermented food or drink is shochu. The liquid koji set produced by the method according to any one of claims 1 to 3 for producing a fermented food or drink having at least glucoamylase and acid-resistant α- amylase .

Images