KR20040049023A - Fermented liquor using purified enzyme and dried koji and preparation method thereof - Google Patents

Abstract

PURPOSE: Fermented liquor using purified enzyme and dried koji and a preparation method thereof are provided, thereby reducing the amount of koji used and maintaining the optimal pH for purified saccharifying enzyme by acid derived from koji without additional regulation of pH, and increasing the fermentation rate and removing distinct smell of koji from the fermented liquor because the dried koji is used as nitrogen source of yeast during the alcohol fermentation. CONSTITUTION: The method for preparing the fermented liquor using purified enzyme and dried koji comprises the steps of: (a) inoculating Aspergillus kawachii(KCCM 11460) into the steamed rice, and heating and drying the steamed rice to prepare the dried koji; (b) adding water and purified liquefying enzyme into the unsteamed grains, liquefying the grains, adding the dried koji and purified saccharifying enzyme into the liquefied grains, and saccharifying the grains; (c) filtering the saccharified grain solution and adding inorganic salts into the filtered solution; (d) sterilizing the inorganic salt added solution; (e) cooling the sterilized solution, inoculating yeast into the solution and fermenting the solution; and (f) filtering the fermented solution, wherein the inorganic salt is selected from ammonium sulfate, ammonium phosphate, potassium phosphate and magnesium sulfate.

Description

Fermented liquor using purified enzyme and dried koji and preparation method

The present invention relates to a fermented liquor using a purified enzyme and a dry country and a method for producing the same. More specifically, the present invention comprises the steps of (a) inoculating Baekkukyun in the cooked rice to prepare a soup, followed by heating and air drying to prepare a dry soup; (b) liquefying by adding water and purified liquefied enzyme to the raw material grains which are not cooked, followed by saccharification by adding the dry country and purified glycosylase of step (a) to the liquefied liquor; (c) filtering the saccharified solution of step (b) and then adding an inorganic salt; (d) sterilizing the saccharified solution to which the inorganic salts of step (c) are added; (e) cooling the sterilizing solution of step (d) and then inoculating the fermentation yeast for fermentation; And (f) filtering and forming the fermentation broth of step (e), and the grain fermented liquor prepared according to the method.

The preference of Takju and Cheongju, which was popular as a popular liquor in Korea, is gradually decreasing. The reason for this is that consumers tend to prefer liquor with relatively simple taste and flavor or low alcohol beer with less hangover burden than grain fermented liquor with complex flavor. Therefore, in order to expand the demand for grain fermented wine, it is necessary to develop fermented wine having a higher preference than conventional Takju or Cheongju.

On the other hand, conventionally, in the manufacturing process of grain fermented wine, such as Takju, Yakju, and Cheongju, yeast or soup have always been added for saccharification of raw grains. However, the addition of yeast or soup has the disadvantage that it gives a complex flavor to the grain fermented liquor, causes a non-uniformity of quality and the process is cumbersome. In order to remedy these disadvantages, a method of using purified enzyme without using yeast or soup was studied when preparing grain fermented liquor. Korean Patent No. 1983-2696 discloses a method of brewing an alcoholic beverage, wherein the steamed starch raw material is glycosylated with saccharifying enzyme and fermented by yeast. Korean Patent No. 1989-4015 discloses grain fermented liquor by fermenting jade powder and malt with proteolytic enzymes, liquefaction enzymes and saccharifying enzymes at high temperature, and then fermenting them by adding yeast to saccharified raw juice. The production method of the other rejuvenation, characterized by the addition of sweeteners and additives is disclosed.

However, there are some problems when the saccharification process is performed using the purified enzyme as described above. First, since the activity of the purified enzyme is high in the low pH range, there is a disadvantage that the pH must be adjusted separately by adding an acidic substance such as lactic acid to the liquefied liquid of the raw grain.

Secondly, when the raw grains are cooked and then saccharified by the purified enzyme, there is a problem in that liquor without fresh flavor is produced due to the heating odor of the cooked raw grains and does not differentiate from conventional grain fermented liquor.

Third, when the raw material grains are glycosylated using only liquefied enzyme or saccharified enzyme, there is a problem in that alcohol fermentation does not occur because of insufficient nitrogen source available for yeast.

In order to solve the above problems, the present inventors add the purified enzyme and the dry soup to the raw grains without increasing the sugar content and saccharify them, and then reduce the total amount of soup used by alcohol fermentation by the yeast, even if no acid is added separately. Due to the acid derived from it, it is possible to maintain the optimum pH condition of the purified glycosylase, and the dry country is used as the nitrogen source of the yeast during the fermentation of alcohol, which improves the fermentation speed and improves the quality. The present invention was completed by preparing flavored grain fermented liquor.

Therefore, the object of the present invention

(a) inoculating Baekkukyun on cooked rice to prepare a soup, and then heating and ventilating to prepare a dry soup; (b) liquefying by adding water and purified liquefied enzyme to raw material grains which are not cooked, and then adding the dry country and purified glycosylase of step (a) to the liquefied saccharification; (c) filtering the saccharified solution of step (b) and then adding an inorganic salt; (d) sterilizing the saccharified solution to which the inorganic salts of step (c) are added; (e) cooling the sterilizing solution of step (d) and then inoculating the fermentation yeast for fermentation; And (f) to provide a method for producing a grain fermented wine comprising the step of filtering and forming the fermentation broth of step (e).

Another object of the present invention is to provide a grain fermented wine produced by the above method.

Figure 1 shows step by step the production method of grain fermented wine according to the present invention.

In order to achieve the above object, the present invention comprises the steps of (a) inoculating Baekkukyun in the cooked rice to produce a soup and then air-dried to prepare a dry soup;

(b) liquefying by adding water and purified liquefied enzyme to the raw material grains which are not cooked, followed by saccharification by adding the dry country and purified glycosylase of step (a) to the liquefied liquor;

(c) filtering the saccharified solution of step (b) and then adding an inorganic salt;

(d) sterilizing the saccharified solution to which the inorganic salts of step (c) are added;

(e) cooling the sterilizing solution of step (d) and then inoculating the fermentation yeast for fermentation; And

(F) provides a method for producing grain fermented wine comprising the step of filtering and forming the fermentation broth of step (e).

In addition, the present invention provides a method for producing grain fermented wine, characterized in that the addition of sugars, acids and medicinal herbs to the fermentation broth of step (e).

The present invention also provides a grain fermented wine produced by the above method.

Hereinafter, the present invention will be described in detail.

Referring to the step-by-step manufacturing method of grain fermented wine according to the present invention.

First step: manufacturing dry country

The dried soup according to the present invention can be prepared by inoculating Bacillus spp. Genus Aspergillus which has excellent organic acid generating ability on cooked rice by empire according to a conventional method. In the present invention, Aspergillus kawachii (KCCM11460) was used as the Bacillus bacterium. The station prepared by the above method is air-dried in a dark room for 1 to 5 days at 40-60 degreeC temperature conditions. By drying the soup prepared as described above, undesirable flavor components affecting the taste of the fermented wine are removed, and there is an advantage that it can be stored for 1 month to 3 months at room temperature. The dried soup is preferably used by grinding with a grinder (blender).

Second step: saccharification of raw grains

To produce the grain fermentation liquor according to the present invention, the raw grains are converted into fermentable sugars available to yeast using purified enzymes.

The raw material grains of the present invention are not particularly limited, and grains and documents containing a lot of starch may be used. Examples of cereals include rice, barley, wheat and corn. Examples of documents include sweet potato starch or potato starch. It is preferable to grind | pulverize with a grinder according to a conventional method, and to use the said raw grain, without increasing.

The saccharification of the raw grains is liquefied using a purified liquefaction enzyme, and then saccharification is performed by adding a purified saccharification enzyme and a drying station according to the present invention.

The liquefaction of the raw material grains was carried out at 90-105 ° C by mixing 10-30% by weight of the raw material grains pulverized without the increase and 70-90% by weight of water with 0.05-1.0% by weight of the purified liquefied enzyme relative to the weight of the raw material grains. Perform for ˜3 hours.

The purified liquefied enzyme randomly hydrolyzes the internal bonds of starch to produce starch suspension by producing small amounts of glucose, maltose, low molecular weight dextrin, and the like which are fermentable sugars available to yeast. It is an enzyme that can convert into a clear solution at high speed. The purified liquefied enzyme is not particularly limited, but α-amylase is representative. Liquefied enzymes can be easily purchased on the market and used in the present invention is termoyl terminus (Termamyl 120L) from Novo, a kind of α-amylase.

The saccharification is performed by adding a dry soup and saccharifying enzyme to the liquefaction of the raw grains obtained by the above method. Preferably, the drying station according to the present invention is glycosylated at 55 to 65 ° C. for 2 to 6 hours by adding 1 to 10% by weight and 0.05 to 1.0% by weight of purified saccharifying enzyme to the weight of the raw grains.

The purified glycosylase refers to an enzyme that is converted into a sugar by hydrolyzing starch partially degraded by liquefied enzyme. The purified glycosylase is not particularly limited, and β-amylase and glucoamylase are typical. The glycosylase can be easily purchased and used commercially, and the one used in the present invention is Novo's AMG E, a kind of glucoamylase.

The glycosylase is generally characterized by high activity in the low pH range. For example, the optimum pH range of the AMG E enzyme purchased from Novo is about 4.1-4.3. Therefore, in order to increase the saccharification efficiency, it is desirable to adjust the optimum pH of the enzyme. In the present invention, when the liquid saccharification of the raw grains using the purified saccharification enzyme, the pH is lowered due to the organic acid derived from the dry country by adding a dry country together to increase the saccharification efficiency. This was confirmed by the present inventors comparing the pH and the reducing sugar content of the saccharification solution prepared by adding a dry country with saccharification enzyme and the saccharification solution prepared without adding the dry country. In other words, the addition of the dry country, the pH was about 4.20, lower than the case without adding the dry country, so that the glycosylation enzyme was an appropriate acidity to show the activity. In addition, it was confirmed that the content of reducing sugar was also higher when the dry country was added than when the dry country was not added.

Step 3: Filtration of Saccharified Liquid and Addition of Inorganic Salts

The saccharified solution prepared in the above step is filtered according to a conventional method, and then inorganic salts are added. Filtration of saccharified liquid can be performed by removing solid content, for example using a filtration tank, a filter, a sachet for filtering sake or a medicine wine, a filter press equipped with the said filter cloth, and a centrifugal dehydrator. At this time, it is preferable to replenish the saccharified liquid by the water lost by evaporation.

In order to promote the growth of fermented yeast, 0.05 to 0.1% by weight of an inorganic salt is added to the filtered saccharified solution. The inorganic salts may be selected from the group consisting of ammonium sulfate, ammonium phosphate, potassium phosphate and magnesium sulfate.

Fourth Step: Sterilization

The saccharified liquid to which the inorganic salt of the said step was added is sterilized at 60-100 degreeC for 10 second-20 minutes. Preferably, it sterilizes 10 minutes-20 minutes at 62-68 degreeC. This is done for the purpose of killing various bacteria in the saccharified solution.

Step 5: fermentation by fermented yeast

The sterilization liquid of the above step is cooled to 15 to 20 ° C., and then fermented with yeast, and fermentation is performed at 12 to 25 ° C. for 5 to 15 days, preferably at 15 to 20 ° C. for 3 to 10 days. Cheongju yeast may be used as the fermented yeast. Preferably, Sacchromyces cerevisiae , a representative sake yeast, is used.

In addition, at this stage, medicinal herbs may be added to enhance the flavor of the fermented wine according to the present invention. Medicines are preferably added at a rate of 0.1 to 5% (w / v) relative to the total fermentation broth between 3 and 10 days after inoculation with yeast and alcohol fermentation. The medicinal herbs can be selected from the group consisting of bokbunja, licorice, wolfberry, Schisandra chinensis and cinnamon, the medicinal herbs are preferably used by grinding.

On the other hand, the inventors of the present invention in order to determine whether the fermentation step by the fermentation yeast dry country is used as a nitrogen source of yeast is enhanced fermentation rate, saccharification liquid prepared using a dry country and saccharification liquid prepared without using a dry country Were prepared respectively and the nitrogen source content contained in the saccharified solution was investigated. In addition, alcohol fermentation was performed using the respective saccharified liquids, and the apparent sugar content of each was measured. As a result, the content of nitrogen source in the case of adding the dry station was very high compared to the case without adding the dry station, and the fermentation rate in the case of adding the dry station also increased significantly compared to the case without adding the dry station. And it was found. It was confirmed that the fermentation rate is larger in the second half. Therefore, the method of manufacturing grain fermented liquor according to the present invention was found to have an effect that the fermentation can be promoted by using the dry country as a nitrogen source of yeast.

Step 6: Filtration and Determination of Fermentation Broth

The fermentation broth of this step is filtered and formulated according to a conventional method. Filtration of the fermentation broth lowers the temperature of the fermenter to -1 to 4 ° C after the fermentation is completed, and precipitates the fermentation yeast and the fermentation residue to obtain only the supernatant. The supernatant obtained is clarified by centrifugation or membrane filter.

Sugars and acids may be further added to the fermented liquor filtered as described above to improve the drinking feeling of the fermented liquor.

Sugars may be added at a rate of 0.5-10% (w / v) relative to the total fermentation broth volume. The saccharide may be selected from the group consisting of glucose, sugar high fructose and starch syrup.

The acids are based on the total fermentation broth volume It can be added at a ratio of 0.1 to 3% (w / v). The acid may be selected from the group consisting of citric acid, malic acid and succinic acid.

The grain fermented wine prepared as described above is bottled or canned according to a conventional method, and then sterilized for 5 to 10 minutes at 62 to 68 ° C. for commercialization.

Grain fermented liquor prepared according to the present invention has a high preference because it has a taste of clean taste and aroma that hardly feels the smell of heat and grain. The present inventors carried out the sensory test of the fermented wine prepared by adding the dry country and the fermented wine produced by adding only the purified enzyme without adding the dry country according to the present invention. As a result, the fermented liquor according to the present invention was excellent in taste, aroma and overall preference.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

<Example 1>

Preparation of Grain Fermented Wine Using Drying Bureau and Purified Enzyme According to the Present Invention

1-1) Manufacturing of Drying Soup

Raw rice was immersed for 3 hours, drained, and cooked. 250 g of cooked rice was inoculated with 0.01 g of Aspergillus kawachii (KCCM11460) spore formation having good organic acid generating ability and incubated for 18 hours at a temperature of 40 ° C. and a humidity of 95%. The station prepared by the above method was air-dried for 2 days in the dark at 45 degreeC temperature conditions. The dried soup was ground finely with a grinder.

1-2) Liquefaction and saccharification of raw grains

250 g of fine powder, 850 ml of water, and 0.2 ml of purified liquefied enzyme (Termamyl, Novo) were placed in a liquefaction vessel and allowed to react for 1 hour while stirring at 100 rpm. After the liquefaction was completed, the temperature of the liquefaction was cooled to 55 ° C, 5 g of the dry soup prepared in Example 1-1) and 0.2 ml of purified glycosylase (AMG E, Novo) were added, followed by reaction for 3 hours.

1-3) Filtration of Saccharified Liquid and Addition of Inorganic Salts

The saccharified solution of 1-2) was replenished with water lost by evaporation and filtered using a filter cloth for sake. To 800 g of the filtered finely divided saccharified solution, 0.02% by weight of ammonium sulfate, 0.02% by weight of divalent ammonium phosphate, 0.015% by weight of divalent potassium phosphate and 0.015% by weight of magnesium sulfate were added.

1-4) Sterilization of Saccharified Liquid and Fermentation with Fermented Yeast

The saccharified solution to which the inorganic salts of Example 1-3) were added was sterilized by heating at 65 ° C. for 5 minutes. After cooling to 15 ° C., about 1.0% (v / v) of Saccharomyces cerevisiae (Japan Brewing Association, Association No. 7) was inoculated and fermented at 20 ° C. for 5 days.

1-5) Filtration and Determination

After the fermentation was completed, the temperature of the fermenter was lowered to 4 ° C. to precipitate the yeast and the fermentation residues, and only the supernatant was obtained. The filtration was clarified by filtration with a membrane filter having a diameter of 1.2 μm. 0.5% citric acid and 1.0% high fructose were added to the circumference prepared as described above, followed by bottling or canning. The grain fermented wine bottled or canned was sterilized at 65 ° C. for 5 minutes to obtain a product.

<Example 2>

Preparation of Grain Fermented Wine Added Cinnamon and Bokbunja According to the Present Invention

The fine powder was saccharified and fermented in the same manner as in Example 1. At the fifth day of fermentation, 0.2% (v / v) of ground cinnamon and bokbunja were added. After the fermentation was completed, the temperature of the fermenter was lowered to 4 ° C. to precipitate yeast, cinnamon, bokbunja, and fermentation residues. The supernatant was taken separately and filtered through a membrane filter having a diameter of 1.2 μm and clarified. Citric acid 0.2% (w / v) and high fructose 0.5% (w / v) were added to the circumference prepared as described above and then bottled or canned. The grain fermented wine bottled or canned was sterilized at 65 ° C. for 5 minutes to obtain a product.

Comparative Example

Preparation of Cereal Fermented Wine without Drying Soup

In the same manner as in Example 1, fine powder was glycosylated using a purified enzyme. At this time, no drying station was added. Yeast was inoculated into the obtained saccharified solution in the same manner as in Example 1, and fermentation was performed.

Experimental Example 1

Comparison of pH, Reducing Sugars and FAN Values with or without Drying Soup

The pH, reducing sugar and FAN (free amino nitrogen) values of the saccharified solution prepared by adding the dry station of Example 1 and the saccharified solution prepared without adding the dry station of Comparative Example 1 were measured and compared. pH was measured using a pH meter (Metrohm, model name 691), the concentration of the reducing sugar was measured by the somogyi (somogyi) method. Meanwhile, the concentration of nitrogen source in the saccharified solution was measured based on free amino nitrogen (FAN) based on the Free amino nitrogen (American Society of Brewing Chemists), the most widely used method of beer analysis.

As a result, as shown in Table 1, the pH of the saccharified solution of Example 1 prepared by adding a dry station was lower than that of Comparative Example 1, which did not add a dry station. The pH of the saccharified solution prepared in Example 1 was measured at 4.20, This is the optimum pH range of the glycosylase (Novo's AMG E) used in the present invention. It was in a range similar to 4.1 to 4.3. Therefore, it was assumed that the addition of dry soup may result in higher glycosylase activity. This estimation was confirmed by measuring the reducing sugar content of the saccharified solution. That is, the reducing sugar content of the saccharified solution prepared by adding the dry country according to the method of the present invention was higher than when the dry country was not added. In addition, the FAN value of the saccharified solution prepared by adding the dry country was higher than that without the dry country. Therefore, the yield of the saccharification process was increased by the addition of the dry country, it can be seen that the nitrogen source significantly increased.

PH, Reducing Sugar, and FAN Values of Saccharified Liquids According to the Addition of Drying Soup Analysis item Comparative example Example 1 pH 5.15 4.20 Reducing Sugar (%, w / v) 15.77 17.14 FAN (ppm) 18.81 129.10

Experimental Example 2

Comparison of Fermentation Rates with or without Dry Soup

In Example 1, the fermentation rate was compared with the case where fermentation was performed using the saccharification solution which did not add the dry country and the saccharification solution prepared without adding the dry country in Comparative Example 1, and the fermentation rate was compared. In order to measure the fermentation rate, the apparent sugar was measured by taking the fermentation broth at 2-day intervals during the fermentation. Apparent sugars were measured with a sugar meter (Density Meter, Anton Paar, model name DMA 4500/5000) using the Apparent Sugar (Apparent Extract) method of the American Society of Brewing Chemists (ASBC). Plato was used as a unit of apparent sugar.

As a result, as shown in Table 2, the case of Comparative Example 1, which does not add a dry station, was significantly slower in change in apparent sugar than in Example 1, which added a dry station. If no dry soup was added, the sugar was not consumed even on the 15th day of fermentation, indicating that fermentation was still in progress. However, in the case of adding dry soup, the sugar was almost consumed on the 9th day of fermentation and completely consumed on the 12th day, indicating that the fermentation rate was significantly increased compared to the case where no dry soup was added. In particular, this gap was larger in the second half of the fermentation. Therefore, it was confirmed that the drying station added during the saccharification process was used as the nitrogen source of the yeast, so that the fermentation rate was increased to improve the fermentation efficiency.

Apparent sugar content Day 0 3rd day 6th day Day 9 Day 12 Day 15 Comparative Example 1 18.10 15.20 11.38 9.44 7.98 6.72 Example 1 18.85 13.81 6.23 2.12 - -

Experimental Example 3

Sensory test of grain fermented liquor without added dry soup and grain fermented liquor according to the present invention

The sensory test of grain fermented liquor which added the dry country of Example 1 and the grain fermented wine which did not add the dry country of Comparative Example 1 was performed. The sensory tests were conducted on 15 well trained test personnel using the Paired Comparison Preference Test. The two-point symbol test is a preference test that determines which of the two specimens is better. The results measured by the 2-point symbol test were verified for significance by the t-test (one-sided test).

As a result, as shown in Table 3, Example 1 with added dry soup was superior to Comparative Example 1 without added dry soup in taste, aroma and overall preference. The numbers shown in Table 3 represent the number of sensory agents preferred for each sample. The results showed that there was a significant difference in the significance level of 1% in the case of fragrance among the items of sensory evaluation, and a significant difference in the significance level of 5% in the case of overall preference. In addition, many of the sensory personnel felt that the fermented liquor of Comparative Example 1 already felt bad and odor due to the fermentation abnormality, while the fermented liquor of Example 1 could not almost feel the smell and feel clean taste and aroma.

Sensory test results of fermented liquor added with dry soup according to the present invention and fermented liquor without added dry soup flavor incense Overall preference Comparative Example 1 5 2 3 Example 1 10 13 12

The method for producing grain fermented liquor according to the present invention has the effect of maintaining the optimum pH condition of the purified glycosylated enzyme due to the acid derived from the dry country even though the amount of the country is reduced, but does not adjust the pH concentration separately. In addition, the drying country is used as a nitrogen source of the yeast during alcohol fermentation, the fermentation rate is increased and the quality is improved, there is an effect that can be produced fermented liquor is given a clean taste and aroma without the smell.

Claims (10)

(a) inoculating Baekkukyun on cooked rice to prepare a soup, and then heating and ventilating to prepare a dry soup; (b) liquefying by adding water and purified liquefied enzyme to the raw material grains which are not cooked, followed by saccharification by adding the dry country and purified glycosylase of step (a) to the liquefied liquor; (c) filtering the saccharified solution of step (b) and then adding an inorganic salt; (d) sterilizing the saccharified solution to which the inorganic salts of step (c) are added; (e) cooling the sterilizing solution of step (d) and then inoculating the fermentation yeast for fermentation; And (F) a method for producing grain fermented wine comprising the step of filtering and forming the fermentation broth of step (e). The method according to claim 1, wherein in step (b), liquefied by adding 0.05 to 1.0% by weight of purified liquefied enzyme to raw material grains to 10 to 30% by weight of raw grains and 70 to 90% by weight of water, and then (a) A method for producing grain fermented liquor characterized by adding 1 to 10% by weight of dry country and 0.05 to 1.0% by weight of purified saccharifying enzyme in step A). The method of claim 1, wherein in step (c), 0.05-0.1 wt% of inorganic salts are added to the saccharification solution of the raw grains. The method of claim 3, wherein the inorganic salts are selected from the group consisting of ammonium sulfate, ammonium phosphate, potassium phosphate and magnesium sulfate. According to claim 1, wherein the fermentation of grain fermented wine characterized in that the addition of 0.1 to 5% (w / v) to the total fermentation broth between 3 to 10 days after the start of fermentation in step (e) Way. The method of claim 5, wherein the medicinal herbs are selected from the group consisting of bokbunja, licorice, wolfberry, schisandra chinensis and cinnamon. According to claim 1, wherein the fermentation broth filtered in the step (f) is a method of producing grain fermented liquor characterized in that further added sugars and acids. The method of claim 7, wherein the sugar is selected from the group consisting of glucose, sugar, high fructose and starch syrup. 8. The method of claim 7, wherein the acid is selected from the group consisting of citric acid, malic acid and succinic acid. A grain fermented liquor prepared by the method of any one of claims 1 to 9.