KR100226201B1 - Preparation of brewing koji - Google Patents

Abstract

The present invention is inoculated with wheat flour at a rate of 15 to 30% by weight, or inoculated with aspergillus Usami or Rizofustelemar spawn, which is a specially cultured seed in yeast raw material obtained by grinding only wheat. (Chopping) The present invention relates to a method for preparing traditional liquor brewing yeast consisting of molding and culturing, and strains of Aspergillus Usami 8731 and Rifupus delemar 1476 used in the preparation. Nuruk manufactured according to the method of manufacturing nuruk of the present invention can be easily mass produced by chopping molding easily and continuously, and it is not inactivated even at pH 2.4 by selecting and culturing the spawn used in the production of nuruk, and has excellent fermentation characteristics when brewing traditional liquor. Can be exercised.

Description

Method of making traditional liquor brewing malt

The present invention relates to a method for producing a traditional liquor brewing nuruk (grain) consisting of sprinkling, seeding and culturing a seed culture cultured on a yeast raw material, and a strain used to prepare the yeast. Specifically, 15 to 30% by weight of wheat flour is mixed with wheat flour, or only wheat is pulverized to prepare a yeast raw material except 80 mesh passage, inoculating pure yeast seedlings with this yeast raw material. The present invention relates to a method for producing a traditional liquor brewing nuruk consisting of culturing after chopping molding to a certain size, and a specific cultured strain used in the manufacturing process of the nuruk.

When only wheat is used, the reason for excluding 80 mesh passes is to adjust the ratio of C (carbon powder) / N (nitrogen powder) to be described later by removing about 10% of the flour component.

Previously, the traditional yeast used in the medicinal liquor factory is a naturally-developed yeast without a specific manufacturing method and is used only in a small amount due to its low titer.In most medicinal liquor shops, aspergillus niger is used for making soju in Japan. Koji is produced by using Aspergillus kawachi, a strain of Aspergillus niger strain, but the burden of manufacturing cost due to low titer This large and often rancid in summer. In addition, since the spawn for producing soju produces organic acids such as oxalic acid, it is effective for producing distilled liquor (ethanol) in hot places such as Japan's Kuju and Okinawa provinces. Another problem is that it is not suitable for brewing. In addition, when these Kawachi bacteria are used, it is cumbersome to incubate under acidic acid by adding acids to koji raw materials.

As an alternative, Korea has developed bran yeast (brand name developing country). However, since the spawn seed for producing the developing country is cultivated in an open manner rather than in a clean room, contamination by wild penicillin (blue mold) is frequently generated and the production is almost stopped. Since the development of technology in this field has progressed since the early 1980s, the seed production was industrialized by pure culture technology using a sealed tank, and research on new yeast production technology is being conducted.

The development of traditional liquor brewing technology requires the improvement of the quality of traditional yeast, and the following problems are still raised in traditional liquor brewing.

-It has low saccharification power and poor quality because it depends on the manufacturing process by natural fermentation.

-Due to the influence of external environmental factors and seasons, there are many differences in quality during manufacturing.

-It is difficult to control the temperature inside the yeast, and the color changes to black or odor due to the propagation of bacteria by abnormal high temperature fermentation.

-Low saccharogenic power (hereinafter referred to as SP), the use rate of yeast is 20 ~ 40% higher than raw materials, and it is difficult to control the quality of alcohol.

-Traditional yeast requires a ripening process to store 1-2 months to improve the quality of yeast, and in reality, the burden of funds from long-term storage is high.

-If the skipping process is omitted, the quality level is lowered, and the low-quality yeast is hard to be drunk.

-For this reason, some conventional yeast and enzymes may be mixed, but this quality is also poor.

-In the manufacture of Takju, Yakju, and Soju, the types of yeast and the conditions of using 3 raw materials should be different.Yakjuyong may have a little color, but Takjuyong must be pure white, so it is white wheat and sojuyong is bran to increase the titer. There should be a lot of ingredients.

The present inventors isolate a specific seed from the tea traditional Nuruk was studied to solve the problems presented above and incubated under specific culture conditions to obtain strains with improved main characteristics as Nuruk for traditional liquor brewing, and then sprinkled with the yeast raw material The present invention was completed by culturing the yeast by culturing the yeast and using the yeast to improve the quality and quality of the traditional liquor.

The present invention, unlike the prior art, by mixing the wheat flour and wheat flour at a ratio of 15 to 30%, or by grinding and selecting only wheat, appropriately control the C (carbon) / N (nitrogen) ratio, here Aspergillus The cultured by adding 40% of Usami's tank-sealed culture and the cultured by adding 40% of the other bacteria, Rizopus delemar, to the above raw materials to make medicine and Takju yeast.

The yeast made by the method of the present invention has a high titer, so that the amount of use can be reduced to at least 1/10. Also, the yeast is molded into a thickness of a boy's finger with a diameter of 8 to 15 mm by a hopper to automatically adjust the room temperature without excessive temperature rise. Can be reduced.

Particularly, in the present invention, the traditional yeast is stored in a well-ventilated place for a long time for 6 months or longer, weevil eats, and the traditional yeast having a titer of 400sp or more is selected from the yeast with the sweet scent, and the bacteria are separated therefrom. Rhizopus genus and Aspergillus genus can be cultured under specific conditions and used as a seed of yeast to improve the taste and aroma of traditional liquor. The mutated strains were identified as Rhizopus delemar strains and Aspergillus usamii strains, and the strains were deposited in the biotechnology laboratory as Rizopus delemar 1476 and Aspergillus usami strains. Registered.

It is an object of the present invention to mix wheat flour in wheat flour at a predetermined ratio within 15 to 30%, or to pulverize only wheat to prepare yeast raw material, and sprinkled and inoculated the seed spawn cultured from the traditional yeast in this raw material and chopper. It is to provide a method for producing a traditional liquor brewing yeast consisting of chopped molding to 9mm size and then cultured.

Another object of the present invention is to provide a strain of Rhizopus delemar 1476 and Aspergillus usamii 8731 which are used to prepare yeast for traditional liquor brewing.

Hereinafter, the present invention will be described in detail.

The manufacturing method of nuruk for brewing traditional liquor according to the present invention is made as follows.

1. In order to prepare yeast raw materials, 15-25 kg of wheat flour is mixed with 100 kg of wheat flour, or only wheat is pulverized.

2. Isolate the Rhizopus delemorr and Aspergillus usamii strains from traditional yeast.

3. Incubate the Aspergillus and Rizopus strains among the isolates.

The spawn culture method is a sterilization method (tank culture), 10% bran medium sterilized at 121 ℃ 1kg / ㎠ for 30 minutes and then cooled to 30 ℃, inoculated with the spawn seed and incubated at 30 ℃ while blowing sterile air . After 48 hours of incubation at pH 2.4 in the first stage, it is expanded for 48 hours at pH 3.0 to 3.5 in two stages.

4. In order to suppress the propagation of germs in nuruk culture, without using water, the spawn cultured by mycelial propagation method (tank culture) is dissolved in water, and the water content is about 95%. Water and inoculate with 100kg of yeast.

5. Using a chopper, it is chopped to a finger thickness (8mm to 15mm).

6. Incubate for 72 hours at 30-40 ℃ in a culture box.

7. Leaving the yeast obtained, it is dried for 20 hours or more at a temperature of 40-50 degreeC.

8. Store and pack for 10-15 days.

The main component of wheat consists of starch and protein. Starch is the main component of carbon (C) component, while protein contains a lot of nitrogen (N). In addition, the flour component contains more starch than protein, and as in the present invention, reducing the wheat flour (flour component) reduces the starch, whose main component is carbon, to increase the nitrogen component of the protein, thereby controlling the C / N ratio. At this time, a large amount of protein components is preferable because the amount of enzyme such as amylase increases. As described above, the present invention can obtain yeast having high saccharification ability in consideration of the C / N ratio. In particular, the sour yeast is irrelevant to the color of liquor (brewing liquor), and when the starch value is high, the saccharification power is poor, so the flour content should be reduced for the purpose of controlling the C / N ratio.

However, the conventional yeast has made the yeast by wheat alone without considering the C / N ratio, so that the enzymatic value of amylase, protease, etc. was lowered. However, the commercially available yeast titer (standard value) is generally recognized to be about 300 sp, but any grain (starch) is known to have this degree of saccharification power. For example, when barley malt and rice are sprayed on the rice paddy, the sensation is due to the amylase that the grain naturally has. Therefore, the amylase titer of the grain itself is added to the yeast of the natural culture method. In addition, even if the amylase of the naturally-occurring fermented yeast on the market is liquefied or saccharified, or acidic or alkaline, the amount of use varies significantly. For example, the amylase titer is almost inactivated when the naturally occurring yeast pH is 3.2 or less, whereas the yeast according to the present invention is not inactivated even at pH 2.4.

In separating the seed from the traditional yeast according to the present invention, the traditional yeast is selected from yeast having a titer of 400sp or more among yeasts that are weeded by the weevil and stored with a fragrant aroma when stored in an airy place for a long period of more than 6 months. .

Aspergillus and Rizopus strains were selected from the isolates, cultured as 10-20 L Jar fermentor by a common microorganism (fungus) culture method, and then inoculated into a seed tank of 1000-2000 L. Incubate at 30 ° C to 35 ° C with stirring for 36 to 48 hours while supplying air. Here, the mushrooms are pulverized with the mold in a liquid sealed container (supernatant 5-10 ~ 20ℓ) in a microorganism propagation incubator 50 mesh (mesh) and put 500g with 5 liters of water in a container, like a container Steam sterilization at 1kg / ㎠ and pressure at 110 ~ 115 ℃ and aseptically cooled, then inoculated with bacteria in an air cleaner to provide constant temperature and sterile air while improving the supply of O ₂ (oxygen) It is a device for obtaining basic data for industrialization on a laboratory scale.

By culturing in a special medium to be described later according to the process as described above it can be obtained strain Rizopus delemar 1476 and strain Aspergillus Usami 8731 according to the present invention.

On the other hand, it will be described in more detail for the microbial strain used to prepare the yeast for brewing traditional liquor according to the present invention.

(1) microbial isolation

Rizopus delemar strain is an acid-resistant Rizopus bacterium, which the inventors applied for in 1968, and the gluco-amylase-producing bacterium was repeatedly isolated from those having higher enzymatic value by UV irradiation, specific soil, or sunlight irradiation. . As a result of the repeated culture, the pure cultured titer of Usami at the beginning of separation was around 1000sp and then increased to 9000 ~ 10000sp, and the pure cultured titer of Lysops was improved to 7000 ~ 8000sp at the initial stage of separation.

These strains degenerate if the separation operation is not continued, and fade from brown when the spores are exposed to sunlight and change to a short spore disease.

The primary superior strains are inoculated with Aspergillus Usami dark brown, and lycopus delemar black or gray with spore color white due to deterioration of color. Wild spores of lycopus species must be culled in dark brown and not pure white.

(2) microbial culture

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The incubator of bacteria used in the production of Nuruk is a culture method that prevents the deterioration of bacteria by using one or two kinds of bovine skin, humus extract and wort or potato juice in a natural medium without using synthetic medium. Continue to disconnect without rest.

The humus extract in the medium is used to maintain or improve the performance of the above-mentioned Rizopus and Aspergillus strains, which are used for at least once a month to cover high altitude corrosive soils that have not been reached by humans for decades. Use at least 50 colonies or more to separate purely and select the highest titer. However, since the bacteria's performance is not fixed and more than 90% of the degenerating strains may be generated, high titers should be selected without any break.

B, culture conditions

Aerobic, temperature 33 ° C, shaking, stationary (liquid)

The developmental temperature is 30-35 degreeC in Aspergillus Usami, and 28-30 degreeC in Rifupus delemar.

All. How Culture

Slant: Incubator 30 ℃

Seed culture: closed tank culture (cultivated by stirring while passing sterile air)

Nuruk cultivation is inoculated and mixed with an amount equivalent to 38 to 40% of the raw material of the incubated seedlings, and then chopped to granule to a predetermined thickness, and the culture temperature is adjusted to 30 to 40 ℃ 72 Incubate time.

(3) microbial microbial properties

Aspergillus Usami 8731 strain of the present invention has strong acid resistance and secretes high titers of gluco-amylase and citric acid.

Rizopus delemar 1476 strain of the present invention secretes a high titer of gluco-β amylase under acidic conditions below the active range of the general microorganism, and produces various organic acids such as fumaric acid, citric acid and lactic acid.

The strain of the present invention has the characteristics of showing good flavor, refreshing taste and umami by secretion of organic acids such as citric acid. Rizopus delemar strains and Aspergillus strains are well known strains.

Rizopus delemar 1476 strain of the present invention is a natural variant Rizopus delemar strain having 1.5 to 2.0 times higher glycosylation properties than the existing Rizopus delemar strain. The glycosylation power of the conventional strain of Rizopus delemar was generally less than 1000 sp (Saccharogenic power), but the strain of Rizopus delemar 1476 of the present invention has a glycosylation power of 3000 to 4000 sp. In addition, the Aspergillus Usami 8731 strain of the present invention is a natural variant Aspergillus Usami strain having a characteristic of 1.2 to 1.5 times stronger than the conventional Aspergillus Usami strain.

Aspergillus Usami 8731 strain of the present invention has a glycosylation potency of 1500-2000sp, as compared to 600-800 sp of the Aspergillus Usami strain.

(4) microbial deposit

As a result of confirming the above strains, it was recognized as belonging to Aspergillus Usami strain and Rizopus Delemar strain, and they were named Aspergillus Usami 8731 and Refupus Delemar 1476, respectively. I have been deposited with my biotechnology institute and have been given accession numbers KCTC 8750P and KCTC 8751P.

The spawn incubated in this way is sprinkled inoculated into the primary mixed yeast raw material by adding the dough water and chopped by using a chopper. In the case of sprinkling inoculation, the amount of spawn for the raw material is preferably 30 to 40%. In the process of sprinkling inoculation, when the spawn seed is moving to the conveyor belt about 3cm thick, inoculate by drilling a large number of 6mm balls from the pipe in a row, inoculating the milled wheat on the belt with a metering pump and passing a mixer rotating between 700 and 800 This can be done evenly. The chopper used in the present invention has a capacity of 7.5 HP.

Choppers are used to incubate the chopped shaped Nuruk raw material to a finger thickness (about 9 mm). The temperature immediately after the incubation is maintained at 30 to 35 ° C, and measures for suppressing moisture divergence from inoculation to 12 to 20 hours are carried out. The high and low temperatures are replaced up and down to keep the temperature of the culture box uniform. Water dissipation can be suppressed by using an ultrasonic humidifier. Ultrasonic humidifiers do not indirectly close the culture.

After that, when the hyphae grows and the white hyphae propagation starts to appear, the temperature rises rapidly, so that the drying prevention cover is appropriately removed in order to prevent such a temperature rise, and at the same time, the temperature of the fermentation chamber is gradually lowered. After about 30 hours, the hyphae will multiply inside the leaven and the calorific value will be the highest and the temperature will rise to 40 ℃. After that, the metabolic heat is gradually lowered, so that the culture box and the box are bonded to each other to prevent rapid moisture dissipation. Normally, after 48 hours, the outland cools, but because it is shaped like a finger, the breeding speed is delayed, but the total incubation time is 72 hours.

As a result of leaving the dried yeast and analyzing the enzyme power according to the National Institute of Technology Research, the yeast according to the traditional method has a titer of 200 to 400sp and a general commercial product of 300sp, whereas the yeast according to the present invention has a titer of 600 to 1500sp. This has been greatly improved.

In the present invention, when producing traditional liquor can be brewed by varying the species. In other words, if you use a mixture of Rizopus and Aspergillus Usami in a 3: 1 ratio, you can obtain a good quality traditional liquor. At this time, if the total amount of the strains used is 2 to 4% with respect to the brewing raw material, sufficient effect can be obtained.

For example, in the case of Japanese sake brewing, the amount of alcohol is the same. ) If you measure the spirit (酒精) up to 21 to 22 degrees after 30 days, yeast according to the present invention can easily reach 19 degrees by fermentation within 10 to 20 days.

The characteristics of the yeast according to the present invention are shown in Table 1 below in comparison with the conventional yeast.

TABLE 1

EXAMPLE

Hereinafter, the present invention will be described in detail with reference to Examples. However, the present invention is not limited by these examples.

Example 1

Spray 100 liters of wheat (mill) with 3 liters of water, and peel the outermost sheath with a peeler (micron) and grind it with a 25 mesh grinder to remove wheat flour with an 80 mesh sieve (10% or so) without kneading with water. After mixing, 20% (15Kg) of wheat flour is mixed with water, and it is dissolved in water and mixed with 70 liters of liquid spawn (Rippus delemar 1476) containing 95% water content, and continuously introduced into a chopper through a convea After extruding about 9mm to about 3cm thick in the culture box and arranged on the incubator shelf, the room temperature is raised to 35 ℃, the temperature is maintained at 30 ~ 35 ℃ 13 to 18 hours, and then the cells of the culture propagated between The ultrasonic humidifier was operated so that the yeast surface did not dry until it was reproduced evenly with the naked eye, and kept the indoor humidity almost saturated. When the mycelium grows in general, the room temperature is not higher than 28 ° C and the moisture temperature is 30 to 35 ° C while supplying moisture for 15 minutes at intervals of about 3 to 4 hours. Adjust to prevent rising for more than 1 ~ 2 hours at the maximum temperature of 40 ℃. After 48 hours, the humidifier is completely shut down and the temperature is maintained between 30 and 33 ℃ until 72 hours. After incubation, transfer to a drying room and blow dry by forced ventilation (hot air 45 ℃). As a result of analyzing the enzyme power of the obtained yeast, it was 690 sp.

Example 2

Nuruk was prepared in the same manner as in Example 1, except that Aspergillus Usami 8731 was used as the liquid seed. The result of enzymatic analysis was 1000 sp.

Example 3

Nuruk was prepared in the same manner as in Example 1, except that Aspergillus Usami strain and Rizopus Delemar strain were mixed at a weight ratio of 1: 1 as a liquid seed. The result of enzymatic analysis was 1000 sp.

[Test Example]

[Comparison of Alcohol Fermentation Tests by Type of Yeast]

According to the National Tax Service liquor analysis regulations, water, yeast and yeast were mixed for 2 hours, as shown in Table 2, and then the yeast was rubbed by hand. Then, white rice was added to the stock. This process is called a one-stage immersion process. After one stage of immersion, it was left for one day, and on the third day, water was poured for two stages of immersion, and rice was added with thickened and cooled rice. The product temperature, pH, acidity, reducing sugar, amino acidity, alcohol specific gravity were measured and the results are shown in Tables 3 to 5 below.

TABLE 2

TABLE 3

TABLE 4

TABLE 5

The Nuruk prepared according to the Nuruk manufacturing method of the present invention is chopped to about 9mm thickness and can be easily molded and mass produced continuously, and it is not inactivated even at pH 2.4 by culturing the spawn used in the manufacture of Nuruk, and is excellent in brewing traditional liquor. Fermentation characteristics can be exerted. In addition, alcohol fermentation stability is great.

Claims (5)

In the method of manufacturing koji for brewing traditional liquor, which is inoculated and incubated with wheat seed, 15-30% of wheat flour is mixed with wheat to prepare a koji ingredient, and Aspergillus usamii 8731 is used as the raw material. (KCTC 8750P) spawn, Rhizopus delemar 1476 spawn (KCTC 8751P) or a mixture thereof is prepared by inoculating water and inoculating sprinkled with 8 to 15 mm in size to incubate brewed yeast. According to claim 1, When the seed inoculated in the yeast raw material is released in water in the state of water content of 95%, the production method of traditional liquor brewing yeast in which the amount of 30 to 40% of the raw material is sprinkled. The method of claim 1, wherein the culture conditions are cultured for 30 hours at 30 ~ 40 ℃ culture sake brewing nuruk. Rhizopus delemar 1476 (Accession No.), which has a glycosylation capacity of 3000 to 4000 sp under acidic conditions of pH 3.2 or less, which is below the active range of general microorganisms, and generates organic acids such as glyco-β amylase and fumaric acid, lactic acid, citric acid, etc. KCTC 8751P) strain. A strain of Aspergillus usamii 8731 (Accession No. KCTC 8750P), which has a glycosylation potency of starch and produces gluco-amylase.