WO2017082499A1 - Mesophilic fermentation yeast and method for preparing same - Google Patents

Mesophilic fermentation yeast and method for preparing same Download PDF

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Publication number
WO2017082499A1
WO2017082499A1 PCT/KR2016/005348 KR2016005348W WO2017082499A1 WO 2017082499 A1 WO2017082499 A1 WO 2017082499A1 KR 2016005348 W KR2016005348 W KR 2016005348W WO 2017082499 A1 WO2017082499 A1 WO 2017082499A1
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yeast
nuruk
fermentation
temperature
fermented
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PCT/KR2016/005348
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French (fr)
Korean (ko)
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여수환
백성열
최한석
강지은
정석태
문지영
백창호
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대한민국(농촌진흥청장)
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Publication of WO2017082499A1 publication Critical patent/WO2017082499A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
    • A23L7/10—Cereal-derived products
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
    • A23L7/10—Cereal-derived products
    • A23L7/104—Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms

Definitions

  • the present invention relates to a medium-temperature fermented yeast and a method of manufacturing the same, and more particularly, to a method of standardizing and producing a yeast with improved quality and improved shelf life.
  • Takju makgeolli
  • medicinal liquor distilled liquor
  • distilled liquor a fermented liquor made by fermenting yeast made by cultivating mold or yeast in cereals. It contains more fiber and bioactive substances than medicinal liquor and distilled liquor.
  • Takju is a traditional Korean liquor, but despite its high nutritional value, it has not been established as a representative liquor. Recently, consumers' desire for liquor diversified, and the tendency to find high-quality liquor has become stronger. Therefore, there is a need for the production of liquor having a high quality and a standardized process.
  • Yeast is made from wheat as its main raw material and fermented by various microorganisms at low temperatures. Through this fermentation process, various enzymes are activated and glycosylation proceeds, resulting in the characteristic taste and aroma of Korean traditional wine (Kim MJ. 2002. The study about tradi t ional Nuruk.Korean J Food Sci Techno 1 9: 32-329).
  • Traditional yeast has a urgent need for standardized product production and quality standardization due to the complex manufacturing process and various useful fermentation microorganisms.
  • it is urgent to study the microbial growth, the scientific control of the metabolic system, and the establishment of a standardized brewing process.
  • Yeast are classified into traditional yeast and modified yeast according to the manufacturing method.
  • Traditionally a variety of microorganisms present in the natural environment are multiplied and made, and the taste and aroma of alcohol produced by saccharification and alcohol fermentation by these strains have a unique advantage.
  • modified yeast the sterilized starch raw material is inoculated with fermented seedlings such as asparagus ( ⁇ s er J / i / s) and lycopus to prevent microbial contamination and to produce a uniform quality and liquor.
  • fermented seedlings such as asparagus ( ⁇ s er J / i / s) and lycopus to prevent microbial contamination and to produce a uniform quality and liquor.
  • Traditional yeast manufacturing process as described above, the flavor of the liquor varies according to the season, there is a disadvantage that it is difficult to manufacture a standard and uniform foam liquor.
  • the present invention is to identify the quality characteristics of the Nuruk according to the fermentation temperature in the traditional process of manufacturing Nuruk, and based on this, the enzyme activity is excellent, establish a nuruk manufacturing technology having a unique taste and aroma to complete the present invention. 3 ⁇ 4.
  • An object of the present invention is to provide a medium-temperature fermented yeast and a method for producing the same.
  • step 2) adding water to the milled mill of step 1) and then infiltrating it;
  • step 2 molding the infiltrated mill of step 2);
  • the present invention provides a method of producing fermented yeast, including the step of drying after fermentation of step 4).
  • the present invention provides a yeast produced by the method for producing medium-temperature fermented yeast of the present invention.
  • the present invention provides a takju prepared by the yeast of the present invention.
  • the present invention relates to a method for producing medium-temperature fermented yeast and rice, and specifically, milled, mixed with water, infiltrated and infiltrated to form nuruk, and fermented by inverting at intervals of 2 days at a temperature of 36 ° C., 45
  • the medium-temperature fermented malt produced by the traditional method by drying at 7 ° C for 7 days has low acidity to prevent microbial contamination, and the low amino acid level is expected to offset the taste felt during preparation of Takju.
  • ⁇ -amylase and glucoamyl ase enzyme activity Excellent glycosylation, organic acid, free amino acid, and fragrance components are analyzed, and the quality of taste and flavor is judged to be excellent.
  • Fermented Nuruk and its manufacturing method use the traditional method to build a high-quality Nuruk manufacturing method, and provide Nuruk It may be useful in the manufacture and production. [Brief Description of Drawings]
  • 1 is a view showing a process of manufacturing a traditional yeast.
  • FIG. 2 is a view showing a commercial Nuruk acid Nuruk and Songhak Nuruk as a comparison.
  • 3 is a view showing the dispersion of the mesophilic fermentation yeast with fermentation period.
  • Figure 4 is a diagram showing the temperature change of fermented yeast and natural fermented yeast.
  • FIG. 5 is a diagram showing a change in physicochemical properties of mesophilic fermented yeast with fermentation period.
  • Figure 5a is a diagram showing the pH change of mesophilic fermentation yeast with fermentation period.
  • Figure 5b is a view showing the acidity change of mesophilic fermentation yeast with fermentation period.
  • 5C is a diagram showing changes in amino acids of different mesophilic fermented yeast during fermentation period.
  • Figure 6 is a diagram showing the change in enzyme activity of mesophilic fermentation yeast with fermentation period.
  • Figure 6a is a graph showing the change of ⁇ -amyl ase activity mesophilic fermentation yeast according to the fermentation time
  • Fig. Figure 6b is a diagram showing the change in glucoamyl ase activity of mesophilic fermented yeast with fermentation period.
  • Figure 6c is a diagram showing the half of ac idi c protease activity of mesophilic fermented yeast with fermentation period.
  • Figure 6d is a diagram showing the change in saccharification power of mesophilic fermented yeast with fermentation period.
  • FIG. 7 is a diagram showing a GC-MS chromatogram of volatile aromatic compounds identified in yeast.
  • 7a is a diagram showing a GC-MS chromatogram of a volatile aromatic compound of acidic yeast.
  • 7B is a diagram showing a GC-MS chromatogram of the volatile aromatic compound of Songhak Nuruk.
  • 7C is a diagram showing a GC-MS chromatogram of volatile aromatic compounds of medium-temperature fermented yeast.
  • FIG. 8 is a view showing the change in enzyme activity of mesophilic fermented Nuruk with storage period.
  • Figure 8a is a diagram showing the change in ⁇ -amyl ase activity of mesophilic fermented yeast with storage period.
  • Figure 8b is a diagram showing the change in glucoamylase activity of mesophilic fermented yeast with storage period.
  • Figure 8c is a diagram showing the change in acidi c protease activity of mesophilic fermented Nuruk with storage period.
  • Figure 8d is a diagram showing the change in saccharification capacity of mesophilic fermented yeast with storage period.
  • step 2) adding water to the milled mill of step 1) and then infiltrating it;
  • step 2 molding the infiltrated mill of step 2);
  • the addition of water in step 2) is preferably from 20 to 30% of hydrous, more preferably from 24 to 28%, and most preferably from 26 to 28%.
  • the molding of step 3) may be molded using a square, a circle, or an oval, and in a specific embodiment of the present invention, the mold is molded using a yeast molding.
  • the fermentation of step 4) is preferably fermented at 34 to 38X: temperature, more preferably at 36 to 37 ° C silver.
  • D In the above method, the fermentation of step 4) is 1 to 30 days . desirable.
  • step 5> of drying it is preferable to dry at 40 to 50 o C
  • the drying is preferably dried for 5 to 10 days, more preferably for 6 to 9 days, most preferably for 7 to 8 days, in a specific embodiment of the present invention, the inventors Grind the wheat, mix water and infiltrate to form Nuruk, then ferment by inverting at 2 days intervals at 36 0 C and drying for 7 days at 45 0 C.
  • the low acidity is effective in preventing microbial contamination, and the low amino acid level is expected to offset the taste felt during preparation of Takju (see Tables 2 and 5), and has excellent ⁇ -amylase and glucoamylase enzyme activity and glycosylation (Tables 3 and 6). ),
  • the organic acid, free amino acid and fragrance components were analyzed to be excellent in taste and aroma quality (see Table 6, Table 8 and Table 10).
  • the middle of the present invention fermented yeast and its manufacturing method can be usefully used as a manufacturing method of high quality Nuruk using the traditional method have.
  • the present invention provides a yeast produced by the method for producing mesophilic fermentation medium of the present invention.
  • the present invention provides a takju prepared by the yeast of the present invention.
  • the yeast is characterized by excellent enzyme activity and shelf life.
  • the Nuruk produced by the method of manufacturing mesophilic yeast is effective for preventing microbial contamination (see Table 2 and FIG. 5), has excellent enzymatic activity and saccharification ability (see Tables 3 and 6), The taste and aroma quality will be unique and excellent ? Judgment (see Table 6, Table 8 and Table 10), by confirming the excellent storage properties (see Table 11 and Figure 8), the mesophilic fermented yeast production method of the present invention can be usefully used for the production of high-quality Nuruk and Takju Can be.
  • the present invention will be described in detail by Examples, Comparative Examples and Experimental Examples.
  • a medium temperature fermented yeast was prepared using the traditional method, and the quality change of the yeast during the manufacturing process was confirmed.
  • Woori wheat (Geumgang wheat) (grown in 2010), grown in Gwangju, Jeonnam, was purchased, and used before grinding to make yeast.
  • Fig. 1 after crushing our wheat into a huller, it was lowered three times with 7-mesh (2.8 mm), and it was allowed to infiltrate at room temperature for 1 hour by spraying a mixture of 26% of hydrolysis.
  • the infiltrated wheat was mixed, weighed 340 g each, and put into a yeast mold.
  • the yeast was molded to the same size and weight by using a molding machine, and then fermented by inverting at 2 days intervals at 36 ° C. After drying for 7 days at 45 0 C using a drying apparatus, fermentation periods 3, 6, 10, 20 and
  • a commercially available acidic yeast of Busan and Songhak nuruk of Gwangju were purchased, and the same quality analysis experiment was performed (FIG. 2). Specifically, acid yeast was purchased from Geumjeong Mt. Makgeolli, and Songhak yeast was purchased from Songhakgok.
  • the weight of yeast was measured with a micro weighing scale ( ⁇ — ⁇ 6000N, CAS Co.) and the size was measured with a Digimatic cl iper (CD-20CPX, Mitutoyo Co.). 50, AND Co.).
  • the change in product temperature during fermentation was measured at 6 hour intervals using the H0B0® company's data logger U12-013 for changes in the leaven temperature during fermentation (0 3, 6 10, 20 30 days).
  • the change in physicochemical properties of the fermented oak was determined by measuring pH ⁇ total acidity (Ac idi ty) and amino acid degree (Amino ac idi ty). Specifically, first, the Nuruk sample for physicochemical characterization was added 100 mL of distilled water to 20 g of the test sample and leached at room temperature for 3 hours to prepare the filtrate solution. Samples for enzyme activity analysis were added to 50 g of sodium chloride solution in 10 g of the sample, shaken occasionally at room temperature (4 ° C) overnight or at room temperature (15-20 ° C) for 3 hours, and then filtered. It was used as an enzyme solution.
  • pH was measured at room temperature with a pH meter (Or ion 3 star, Thermo scienti Co Co.). The total acidity was determined by taking a certain amount of sample, taking 10 mL of the sample filtered with filter paper into a 100 mL Erlenmeyer flask, dropping 2-3 drops of 0.5% phenolphthalein indicator and neutralizing titration until it became red with 0.1 N NaOH. The amount of the prepared solution was converted into lactic acid (%).
  • the acidity of the mesophilic fermented yeast produced was 2 to 5 times lower than that of commercial Nuruk, which was expected to be effective in preventing contamination of various microorganisms such as harmful microorganisms in the early stage of Nuruk fermentation.
  • acidic Nuruk and Songhak Nuruk were the same value as 1.28 mL, and the intermediate temperature fermented Nuruk was increased in amino acidity with fermentation period. It decreased to 0.48 mL with fermentation period from 10 days of fermentation (Table 2 and Fig. 5). . Therefore, since the amino acidity of the produced medium-temperature fermented yeast is about three times lower than the commercial Nuruk used as a control, the taste of the aftertaste will be canceled much when Takju or Yakju is prepared. This may affect the change of taste.
  • Control (commercial Nuruk) Acid 6.64 ⁇ 0.03 0.01881 ⁇ 0.0068 1.28 ⁇ 0. 10
  • Control Common Yeast) Transfer 6.29 ⁇ 0.02 0.1518 + 0.0143 1.28 ⁇ 0.01
  • a-amylase activity 50 mL of sodium chloride solution was added to 10 g of yeast, and the solution was sometimes shaken at low temperature overnight or at room temperature for 3 hours, and then the filtrate was diluted to prepare a crude enzyme solution. Take 2 mL of 1% starch solution into the test tube, preheat it at 40 ° C for 5 minutes, add 0.1 mL of enzyme solution to start reaction, and pipette 0.01 mL of iodine solution into the reaction solution in 1 minute intervals. Was added to a 10 mL test tube to maintain the resulting mixture at 25 ° C. After passing through a thick 10 mm to compare the 670 nm color, 1% transmittance was measured. Enzyme activity (uni t) was calculated by the following Equation 1 based on the Wohlgemuth value.
  • T 0 rain Permeability before enzyme reaction
  • Acidi c protease activity was determined by adding 50 mL of sodium chloride solution to 10 g of yeast feed. After leaching overnight in a low temperature room (4 ° C.) or at room temperature (15-20 ° C.) for 3 hours, it was filtered. 10 mL of the filtrate was placed in a dialysis membrane and dialyzed overnight at 4 ° C. in 10 to 2 M acetic acid buffer, and the enzyme solution was diluted to form a crude enzyme solution. After adding 1.0 mL of pH 3.0 McVine buffer solution and 0.5 mL of coenzyme solution to 1.5 mL of casein solution, the mixture was reacted at 40 ° C.
  • Glucoamylase activity was preheated for 5 minutes at 40 ° C by adding 0.2 mL of 0.2 M acetic acid buffer to 1 mL of 2% starch solution, and adding 0.1 mL of yeast enzyme solution and reacting for 20 minutes at 40 o C. The reaction was stopped by the addition of 0.01 mL of NaOH solution, left for 30 minutes, and neutralized by addition of 0.01 mL of 1 N hydrochloric acid solution. The control group used in the above experiment was preheated at 40 ° C. for 5 minutes by adding 0.2 mL of 0.2 M acetic acid buffer to 1 mL of starch solution.
  • the control group was neutralized by adding 0.01 mL of 1 N hydrochloric acid solution in the same manner as the sample, and the enzyme activity was the amount of glucose produced by measuring the reducing sugar by DNS method.
  • Glucoamylase activity was 1 uni t to produce 1 mg and glucose for 60 minutes at 40 o C from soluble starch, glucoamylase activity of the sample 1 g was calculated by the following equation (3).
  • the yeast enzyme solution was diluted by 2 dilutions, at least 10 mL, and used as a coenzyme solution.
  • 50 mL of 2% starch solution and 30 mL of vinegar acid buffer solution were taken in a 100 mL volumetric flask, preheated in a constant temperature water bath preheated to 55 0 C for 10 minutes, and 10 mL of enzyme solution was added and saccharified for 60 minutes.
  • the control was added to 10 mL of distilled water and saccharified the same for 60 minutes.
  • 10 mL of 0.5 N NaOH was added thereto, the enzyme reaction was stopped, and rapidly cooled, and then adjusted to 100 mL by adding distilled water.
  • the ⁇ -amylase activity was 175.1 and 280.5 units / g of commercial yeast of ⁇ Comparative Example 2> as a control.
  • Medium-temperature fermented Nuruk (TN-A) increased its activity with fermentation period, and a-amylase activity was 310.3 units / g, 1.2 times higher than that of commercial Nuruk (Table 3 and Figure 6).
  • the activity of glucoamylase was 908 and 3,372 units / g in acidic yeast and Songhak yeast, and the temperature of fermented yeast decreased with increasing fermentation period.
  • the value of 5,592 units / g was higher than that of control. It showed 2-3 times higher activity (Table 3 and FIG. 6).
  • the saccharification power of the mesophilic fermented yeast prepared in Example 1 is higher than that of commercial Nuruk, which indicates that the effect of decomposing rice starch into sugars by enzymatic activity of raw grain an Nuruk is great.
  • the acidic protein activity was 4,779.8 and 4,212.5 units / g in acidic Nuruk and Songhak Yellow, and the temperature of fermented Nuruk increased with increasing fermentation time.
  • the enzyme activity was 1,928.5 units / g at 30 days of fermentation. It was about 3 times lower than commercial leaven (Table 3 and FIG. 6).
  • the yeast is so proteolytically high enough that the meju and its components are different, the protein is broken down and converted into amino acids. At this time, since the taste difference of the main is determined, it is advantageous to have low proteolytic power.
  • the medium-temperature fermented yeast prepared in ⁇ Example 1> and commercial Nuruk (acidic Nuruk and Songhak Nuruk) and organic acid of ⁇ Comparative Example 2> were analyzed.
  • citric acid (citric acid) was detected as a major organic acid in the acidic malt which is a commercial malt of ⁇ Comparative Example 2>, and Songhak malt was citric acid, malic acid (apple acid) and lactic. acid (lactic acid) was analyzed as the main organic acid (Table 6).
  • citric acid, malic acid, and oxalic acid were analyzed, and citric acid (citric acid) increased as early organic fermentation and kept constant from 10 days (Table 6).
  • the amount and type of organic acid differed by yeast, which was considered to be different according to raw materials, related fermentation microorganisms and fermentation conditions.
  • the main amino acids of commercial yeast in Comparative Example 2 were proline (sweet / bitter), alanine (sweet), valine (light bitter), glutamic acid (mild), ⁇ - It was composed of 14 free amino acids such as aminobutyric acid, and the total amino acid content was 602.92 / g / mL, and the essential amino acid content was 21.14 / g / mL.
  • the major free amino acids of Songhak Nuruk consisted of 24 free amino acids including proline (sweet / bitter), alanine (sweet), valine (weak bitter), urea, ⁇ -aminobutyr ic acid, and the total amino acid content is 929.98 / g / mL, which is required The amino acid content was 103.63 / g / m £ (Table 8).
  • Fermentation 30 primary key amino acids of mesophilic fermenting yeast of the invention prol ine (sweet / bitter taste), val ine (weak bitter taste), taur ine, alanine, ⁇ -aminobutyr ic acid such as' was composed of eight yungri amino acids, a total of The amino acid content was 107.23 g / mL, of which the amino acid content was 12.27 / g / mL (Table 8).
  • HP-INN0WAX 60 m ⁇ ⁇ .25 mm i, d ⁇ ⁇ .25 pm film thikness
  • Injection temperature 250 ° C
  • a total of 48 aroma components were detected, including 10 species, 4 types of acid series, and 2 types of alco) l.
  • the common fragrance components of the three types of yeast were octane, formic acid ethyl ester, (E) -6-met hy 1-3-undecene, decane, methyl benzene and undecane It was composed of 12 components, metyhl laurate, 1,3 'dimethyl-benzene, 1, 2-dimethyl-benzene, l_ethyl_3' methyl ⁇ benzene, acidic acid and n-hexadecanoin acid (Table 10).
  • yeast is the yeast acid (9Z, 12Z) -9, 12 -octadecadienoate ethyl, ethyl palmitate, n-hexadecanoic acid, 8-heptadecene, 9-octadecenoic acid species such as 22 to "ingredient songhak yeast is acetic acid .27 ' components such as ethyl ester, n-hexadecanoic acid, octadec-9-enoic acid, methyl octadeca-9, 12-dienoate, 1-tr icosene, etc.
  • TN_A Medium fermented yeast
  • n -hexadecanoic acid 4-ethoxy- benzoic acid ethyl ester, 1, 3-di me t hy 1 -benzene, 2-methyl -octane such species i, was analyzed by 25 component (Table 10).
  • Table 10 The composition of the fragrance between commercial Nuruk and the elevated temperature fermented Nuruk of the present invention does not show a big difference.
  • Zb hexadecanoic acid was the most important component of yeast with more than 30 ⁇ 40% .
  • Acetic acid ethyl ester is a natural component of fragrance that exists in fruits such as pineapple. It is a fragrance ingredient.
  • n-hexadecanoic acid is a waxy, soft and sweet fragrance that is recognized as a fragrance ingredient.
  • Ethyl ester is an ingredient of soy sauce that has been analyzed as medium fermented malt and characteristic flavor component.
  • Experimental Example 9 confirms the shelf life of fermented yeast
  • ⁇ Example 1> During the preparation in ⁇ Example 1> was analyzed the remaining capacity of the yeast enzyme according to the storage period of the fermented yeast. ' Specifically, ⁇ -amylase, glucoamylase, acidic protease activity and .
  • the glycosylation potency (SP) was measured, and was carried out in the same manner as in ⁇ Example 3>, and the residual ability of the yeast enzyme at 4 ° C., 15 ° C. and 30 ° C. during 0, 8 and 12 weeks storage period was analyzed. It was. As a result, as shown in Table 11 and FIG.
  • Acidic c protease activity analysis showed that the 12-week enzyme activity of mesophilic fermented yeast was maintained at 94.8% with 2, 776.3 uni ts / g at 4 0 C storage condition. At 15 ° C storage condition, 92.5% was maintained at 2, 708.8 uni ts / g, the actual 7.5% enzyme was inactivated, and at 8 ° C at 30 ° C storage condition was maintained at 29.8% 9.9 ts / g. 10.2% of the enzyme was inactivated (Table 11 and FIG. 8).

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Abstract

The present invention relates to a mesophilic fermentation yeast and a method for preparing the same. More particularly, the mesophilic fermentation yeast is prepared in a traditional manner which comprises the steps of crushing wheat, mixing the crushed wheat with water, wetting the mixture to form yeast, fermenting the yeast while flipping the same in intervals of two days at a temperature of 36°C, and drying the same at 45°C for seven days. The mesophilic fermentation yeast has been observed to have a low level of acidity, and thus is effective in preventing microbial contamination, have a reduced level of amino acidity, and thus is considered to offset a greasy taste upon preparation of rice wine, and have an excellent α-amylase and glucoamylase enzymatic activity, excellent saccharification power, excellent qualities of taste and fragrance as a result of analyzing free acid, free amino acid and aroma components, and an excellent retention capacity of an enzyme depending on a storage period, thereby showing excellent storability. The mesophilic fermentation yeast and the method for preparing the same of the present invention allow a method for preparing yeast having an excellent quality to be established by using the traditional manner, and can be usefully employed in the preparation and production of yeast by providing the yeast prepared therefrom.

Description

【명세서】  【Specification】
【발명의 명칭】  [Name of invention]
중온 발효누룩 및 이의 제조방법 [기술분야】 Medium-temperature fermented yeast and preparation method thereof
본 발명은 중온 발효누룩 및 이의 제조방법에 관한 것으로， 구체적으로 품질이 개선되고, 저장성이 향상된 누룩을 규격화하여 제조할 수 있는 방법에 관한 것이다. The present invention relates to a medium-temperature fermented yeast and a method of manufacturing the same, and more particularly, to a method of standardizing and producing a yeast with improved quality and improved shelf life.
【배경기술】 Background Art
세계와 각국에는 그 지역의 기후， 풍토 및 습관에 따른 지방 고유의 술, 즉 민속주， 전통주 또는 지역 특산주라는 특유의 술이 전해져 오고 있다. 우리나라의 경우 탁주 (막걸리)， 약주 및 증류주 등으로 불라는 전통주가 오래 전부터 이어져 내려오고 있다. 탁주 (막걸리)는 곰팡이나 효모를 곡류에 배양시켜 만든 누룩을 이용하여 발효시켜 만든 발효주의 일종으로， 약주 및 증류주보다 식이섬유 및 생리활성 물질이 다량으로 함유되어 있어， 영양학적 가치가 높다. In the world and around the world, local sake according to the climate, climate and habits of the region, namely sake, traditional sake or local sake, is delivered. In Korea, traditional liquor called Takju (makgeolli), medicinal liquor, and distilled liquor has been passed down for a long time. Takju (makgeolli) is a fermented liquor made by fermenting yeast made by cultivating mold or yeast in cereals. It contains more fiber and bioactive substances than medicinal liquor and distilled liquor.
앞서 말한 바와 같이 탁주는 우리나라의 전통주로 영양학적 가치가 높음에도 불구하고， 대표적인 주류로 자리 잡지 못하고 있는 실정이며， 최근 주류에 대한 소비자의 욕구가 다양화되고， 고품질의 주류를 찾는 경향이 강해짐에 따라 우수한 품질을 갖추고 표준화된 공정을 갖는 주류 제조에 필요성이 대두되고 있다. 누룩은 밀을 주원료로 성형하여 저온쎄서 다양한 미생물에 의해 발효되며, 이와 같은 발효 과정을 거쳐 다양한 효소가 활성화되고 당화가 진행되어 한국 전통 주의 특징적인 맛과 향을 내게 된다 (Kim MJ . 2002. The study about tradi t ional Nuruk. Korean J Food Sci Techno 1 9： 324-329) . 전통누룩은 제조공정이 복잡하고 유용 발효미생물이 다양하여 규격화된 제품 생산과 품질 표준화가 시급한 실정이다. 국내 전통누룩업체에서는 자연발효와 경험에 의존한 누록제조로 미생물 생육과 대사계의 과학적 조절과 표준화된 양조공정 확립이 제대로 이루어지지 않아 이에 대한 연구 또한 시급히 이루어져야 한다. As mentioned before, Takju is a traditional Korean liquor, but despite its high nutritional value, it has not been established as a representative liquor. Recently, consumers' desire for liquor diversified, and the tendency to find high-quality liquor has become stronger. Therefore, there is a need for the production of liquor having a high quality and a standardized process. Yeast is made from wheat as its main raw material and fermented by various microorganisms at low temperatures. Through this fermentation process, various enzymes are activated and glycosylation proceeds, resulting in the characteristic taste and aroma of Korean traditional wine (Kim MJ. 2002. The study about tradi t ional Nuruk.Korean J Food Sci Techno 1 9: 32-329). Traditional yeast has a urgent need for standardized product production and quality standardization due to the complex manufacturing process and various useful fermentation microorganisms. In the traditional yeast company in Korea, it is urgent to study the microbial growth, the scientific control of the metabolic system, and the establishment of a standardized brewing process.
누룩은 제조 방법에 따라 전통누특과 개량누룩으로 분류된다. 전통누록은 자연 환경 중에 존재하는 다양한 미생물이 증식되어 만들어지는 방식이며， 이러한 균주에 의해 당화와 알코올 발효에 의해 제조된 술의 맛과 향이 독특한 장점이 있다. 개량누룩의 경우， 잡균 오염을 방지하고， 균일한 품질와 술을 제조하기 위해 살균한 전분질 원료에 아스파질러스 (^s er J/i/s)와 라이조푸스 와 같은 발효종균을 접종하여 만든다. 전통적 누룩 제조공정은 앞서 말한 바와 같이 술의 풍미가 계절에 따라 다양하여， 표준화되고 균일한폼질의 술을 제조하기 어려운 단점이 있다. 시판누룩은 품질이 열악한 관계로 주질 열악 등 우리술의 경쟁력 또한 낮은 상태이다. 이를 극복하기 위해, 누룩의 품질 개선과 고급화 및 저장성 향상 등의 과학화 연구를 통해， 우리술의 향미에 영향을 주는 효모， 유산균 등 발효미생물을 이용함으로써 우리술에 맞는 누룩제조기술의 선점 연구가 무엇보다도 필요하다. Yeast are classified into traditional yeast and modified yeast according to the manufacturing method. Traditionally, a variety of microorganisms present in the natural environment are multiplied and made, and the taste and aroma of alcohol produced by saccharification and alcohol fermentation by these strains have a unique advantage. In the case of modified yeast, the sterilized starch raw material is inoculated with fermented seedlings such as asparagus (^ s er J / i / s) and lycopus to prevent microbial contamination and to produce a uniform quality and liquor. Traditional yeast manufacturing process, as described above, the flavor of the liquor varies according to the season, there is a disadvantage that it is difficult to manufacture a standard and uniform foam liquor. Due to the poor quality of commercial Nuruk, our liquor's competitiveness such as poor quality is low. In order to overcome this problem, what is the preemptive research of yeast manufacturing technology that is suitable for Korean wine by using fermented microorganisms such as yeast and lactic acid bacteria that affect the flavor of Korean wine through scientific research such as improving the quality of the yeast and improving the quality and shelf life. Than is necessary.
이에, 본 발명은 전통방식의 누록 제조공정 중에서 발효 온도에 따른 누룩의 품질 특성을 규명하고, 이를 바탕으로 효소 활성이 우수하고， 독특한 맛과 향을 갖는 누룩제조 기술을 확립하여 본 발명을 완성하 ¾다. Accordingly, the present invention is to identify the quality characteristics of the Nuruk according to the fermentation temperature in the traditional process of manufacturing Nuruk, and based on this, the enzyme activity is excellent, establish a nuruk manufacturing technology having a unique taste and aroma to complete the present invention. ¾.
【발명의 상세한 설명】 [Detailed Description of the Invention]
【기술적 과제】 본 발명의 목적은 중온 발효누룩 및 이의 제조방법을 제공하는 것이다. [Technical problem] An object of the present invention is to provide a medium-temperature fermented yeast and a method for producing the same.
【기술적 해결방법] Technical Solution
상기 목적을 달성하기 위하여， 본 발명은， In order to achieve the above object, the present invention,
1) 밀을 분쇄하는 단계; 1) milling the wheat;
2) 상기 단계 1)의 분쇄된 밀에 물을 첨가한 후 침윤시키는 단계;  2) adding water to the milled mill of step 1) and then infiltrating it;
3) 상기 단계 2)의 침윤시킨 밀을 성형하는 단계;  3) molding the infiltrated mill of step 2);
4) 상기 단계 3)의 성형된 밀을 34내지 38°C 온도에서 발효시키는 단계; 및 4) fermenting the shaped mill of step 3) at a temperature of 34 to 38 ° C .; And
5) 상기 단계 4)의 발효 후 건조시키는 단계를 포함하는 중은 발효누룩의 제조방법을 제공한다. 또한， 본 발명은 본 발명의 중온 발효누룩 제조방법으로 제조된 누룩을 제공한다. 또한， 본 발명은 본 발명의 누룩으로 제조된 탁주를 제공한다. [발명의 효과] 5) provides a method of producing fermented yeast, including the step of drying after fermentation of step 4). In addition, the present invention provides a yeast produced by the method for producing medium-temperature fermented yeast of the present invention. In addition, the present invention provides a takju prepared by the yeast of the present invention. [Effects of the Invention]
본 발명은 중온 발효누룩 및 미의 제조방법에 관한 것으로， 구체적으로 밀을 분쇄하여， 물을 흔합한 후 침윤시켜 누룩을 성형한 후， 36°C 온도에서 2일 간격으로 뒤집기하며 발효시키고， 45°C에서 7일 동안 건조시켜 전통방식으로 제조된 중온 발효누룩이 산도가 낮아 미생물 오염방지 효과적이고， 아미노산도가 낮아 탁주 제조시 느끼한 맛이 상쇄될 것으로 판단되며， α-amylase , glucoamyl ase 효소활성과 당화력이 뛰어나고， 유기산, 유리아미노산 및 향기 성분을 분석한 결과 맛과 향의 품질이 우수할 것으로 판단되며， 저장 기간에: 따른 효소의 잔존능이 우수하여 저장성이 뛰어남을 확인함으로써， 본 발명의 중온 발효누룩 및 이의 제조방법은 전통방식을 이용하여 우수한 품질의 누룩제조 방법을 구축하고， 이로 제조한 누룩을 제공하여 누룩제조 및 생산에 유용하게 이용될 수 있다. 【도면의 간단한 설명】 The present invention relates to a method for producing medium-temperature fermented yeast and rice, and specifically, milled, mixed with water, infiltrated and infiltrated to form nuruk, and fermented by inverting at intervals of 2 days at a temperature of 36 ° C., 45 The medium-temperature fermented malt produced by the traditional method by drying at 7 ° C for 7 days has low acidity to prevent microbial contamination, and the low amino acid level is expected to offset the taste felt during preparation of Takju. Α-amylase and glucoamyl ase enzyme activity Excellent glycosylation, organic acid, free amino acid, and fragrance components are analyzed, and the quality of taste and flavor is judged to be excellent. During storage period: Fermented Nuruk and its manufacturing method use the traditional method to build a high-quality Nuruk manufacturing method, and provide Nuruk It may be useful in the manufacture and production. [Brief Description of Drawings]
도 1은 전통누룩의 제조 과정을 나타낸 도이다. 1 is a view showing a process of manufacturing a traditional yeast.
도 2는 비교구로서 시판누룩인 산성누룩과 송학누룩을 나타낸 도이다. 2 is a view showing a commercial Nuruk acid Nuruk and Songhak Nuruk as a comparison.
도 3은 발효 기간에 따른 중온 발효누룩의 변화흩 나타낸 도이다. 3 is a view showing the dispersion of the mesophilic fermentation yeast with fermentation period.
도 4는 중은 발효누룩 및 자연 발효누룩의 품온 변화를 나타낸 도이다. Figure 4 is a diagram showing the temperature change of fermented yeast and natural fermented yeast.
도 5는 발효 기간에 따른 중온 발효누룩의 이화학적 특성 변화를 나타낸 도아다. 도 5a 발효 기간에 따른 중온 발효누룩의 pH 변화를 나타낸 도이다. 5 is a diagram showing a change in physicochemical properties of mesophilic fermented yeast with fermentation period. Figure 5a is a diagram showing the pH change of mesophilic fermentation yeast with fermentation period.
도 5b는 발효 기간에 따른 중온 발효누룩의 산도 변화를 나타낸 도이다. Figure 5b is a view showing the acidity change of mesophilic fermentation yeast with fermentation period.
도 5c는 발효 기간에 ！다른 중온 발효누룩의 아미노산도 변화를 나타낸 도이다. 5C is a diagram showing changes in amino acids of different mesophilic fermented yeast during fermentation period.
도 6는 발효 기간에 따른 중온 발효누룩의 효소활성 변화를 나타낸 도이다. Figure 6 is a diagram showing the change in enzyme activity of mesophilic fermentation yeast with fermentation period.
도 6a는 발효 기간에 따른 중온 발효누룩의 α-amyl ase 활성 변화를 '나타낸 도이다. 도 6b는 발효 기간에 따른 중온 발효누룩의 glucoamyl ase 활성 변화를 나타낸 도이다. Figure 6a is a graph showing the change of α-amyl ase activity mesophilic fermentation yeast according to the fermentation time, Fig. Figure 6b is a diagram showing the change in glucoamyl ase activity of mesophilic fermented yeast with fermentation period.
도 6c는 발효 기간에 따른 중온 발효누룩의 ac idi c protease 활성 반화를 나타낸 도이다. Figure 6c is a diagram showing the half of ac idi c protease activity of mesophilic fermented yeast with fermentation period.
도 6d는 발효 기간에 따른 중온 발효누룩의 당화력 변화를 나타낸 도이다. Figure 6d is a diagram showing the change in saccharification power of mesophilic fermented yeast with fermentation period.
도 7은 누룩에서 확인 된 휘발성 향기 화합물의 GC-MS 크로마토그램을 나타낸 도이다. 7 is a diagram showing a GC-MS chromatogram of volatile aromatic compounds identified in yeast.
도 7a는 산성누룩의 휘발성 향기 화합물의 GC-MS 크로마토그램을 나타낸 도이다. 도 7b는송학누룩의 휘발성 향기 화합물의 GC-MS 크로마토그램을 나타낸 도이다. 도 7c는 중온 발효누룩의 휘발성 향기 화합물의 GC-MS 크로마토그램을 나타낸 도이다. 7a is a diagram showing a GC-MS chromatogram of a volatile aromatic compound of acidic yeast. 7B is a diagram showing a GC-MS chromatogram of the volatile aromatic compound of Songhak Nuruk. 7C is a diagram showing a GC-MS chromatogram of volatile aromatic compounds of medium-temperature fermented yeast.
도 8은 저장 기간에 따른 중온 발효누룩의 효소활성 변화를 나타낸 도이다. 8 is a view showing the change in enzyme activity of mesophilic fermented Nuruk with storage period.
도 8a는 저장 기간에 따른 중온 발효누룩의 α-amyl ase 활성 변화를 나타낸 도이다. 도 8b는 저장 기간에 따른 중온 발효누룩의 glucoamylase 활성 변화를 나타낸 도이다. Figure 8a is a diagram showing the change in α-amyl ase activity of mesophilic fermented yeast with storage period. Figure 8b is a diagram showing the change in glucoamylase activity of mesophilic fermented yeast with storage period.
도 8c는 저장 기간에 따른 중온 발효누룩의 acidi c protease 활성 변화를 나타낸 도이다. Figure 8c is a diagram showing the change in acidi c protease activity of mesophilic fermented Nuruk with storage period.
도 8d는 저장 기간에 따른 중온 발효누룩의 당화력 변화를 나타낸 도이다. [발명의 실시를 위한 최선의 형태] 이하본 발명을 상세히 설명한다. 본 발명은, Figure 8d is a diagram showing the change in saccharification capacity of mesophilic fermented yeast with storage period. BEST MODE FOR CARRYING OUT THE INVENTION The present invention is described in detail below. The present invention,
1) 밀을 분쇄하는 단계;  1) milling the wheat;
2) 상기 단계 1)의 분쇄된 밀에 물을 첨가한 후 침윤시키는 단계;  2) adding water to the milled mill of step 1) and then infiltrating it;
3) 상기 단계 2)의 침윤시킨 밀을 성형하는 단계;  3) molding the infiltrated mill of step 2);
4) 상기 단계 3)의 성형된 밀을 34 내지 380C 온도에서 발효시키는 단계; 및 4) fermenting the shaped mill of step 3) at a temperature of 34 to 38 0 C; And
5) 상기' 단계 4)의 발효 후 건조시키는 단계를 포함하는 중온 발효누룩의 제조방법을 제공한다 5) provides a process for the preparation of mesophilic fermentation yeast, comprising the step of fermentation and then drying of the Step 4)
상기 방법에 있어서， 단계 2)의 물 첨가는 가수량 20 내지 30% 인 것이 바람직하며， 24 내지 28%인 것이 더욱 바람직하며， 26 내지 28%인 것이 가장 바람직하다. In the above method, the addition of water in step 2) is preferably from 20 to 30% of hydrous, more preferably from 24 to 28%, and most preferably from 26 to 28%.
상기 방법에 있어서， 단계 3)의 성형은 사각형， 원형， 또는 타원형을 이용하여 성형할 수 있으며， 본 발명의 구체적인 실시예에서는 누룩 성형 를을 사용하여 성형하였다. In the above method, the molding of step 3) may be molded using a square, a circle, or an oval, and in a specific embodiment of the present invention, the mold is molded using a yeast molding.
상기 방법에 ¾어서， 단계 4)의 발효는 34내지 38X: 온도에서 발효시키는 것이 바람직하고， 36 내지 37°C 은도에서 발효시키는 것이 더욱 바람직하다. 또한 상기 발효 기간 중 2 내지 4일 간격으로 뒤집으며 발효시키는 것이 바람직하다. D 상기 방법에 있어서, 단계 4)의 발효는 1일 내지 30일인 것이 .바람직하다. Following the process, the fermentation of step 4) is preferably fermented at 34 to 38X: temperature, more preferably at 36 to 37 ° C silver. In addition, it is preferable to invert the fermentation at intervals of 2 to 4 days during the fermentation period. D In the above method, the fermentation of step 4) is 1 to 30 days . desirable.
상기 방법에 있어서, 단계 5>의 건조는 40 내지 50oC 은도에서 건조시키는 것이 바람직하며， 42 내지 48°C인 것이 더욱 바람직하며， 45 내지 47°C인 것이 가장 바람직하다. 또한 상기 건조는 5 내지 10일 동안 건조시키는 것이 바람직하며， 6 내지 9일 동안 건조시키는 것이 더욱 바람직하고， 7 내지 8일 동안 건조시키는 것이 가장 바람직하다， 본 발명의 구체적인 실시예에서， 본 발명자는 밀을 분쇄하여， 물을 흔합한 후 침윤시켜 누룩을 성형한 후， 360C 온도에서 2일 간격으로 뒤집기하며 발효시키고, 450C에서 7일 동안 건조시켜 전통방식으로 제조된 중온 발효누룩이 산도가 낮아 미생물 오염방지 효과적이고， 아미노산도가 낮아 탁주 제조시 느끼한 맛이 상쇄될 것으로 판단되며 (표 2 및 도 5 참조)， α-amylase , glucoamylase 효소활성과 당화력이 뛰어나고 (표 3 및 도 6 참조)， 유기산， 유리아미노산 및 향기성분을 분석한 결과 맛과 향의 품질이 우수할 것으로 판단되며 (표 6， 표 8 및 표 10 참조)， 저장 기간에 따른 효소의 잔존능이 우수하여 저장성이 뛰어남을 확인함으로써 (표 11 및 도 8 참조)， 본 발명의 중은 발효누룩 및 이의 제조방법은 전통 방식을 이용하여 우수한 품질의 누룩 제조방법으로 유용하게 이용될 수 있다. 또한， 본 발명은 본 발명의 중온 발효누록 제조방법으로 제조된 누룩을 제공한다. 또한， 본 발명은 본 발명의 누룩으로 제조된 탁주를 제공한다. In the above method, step 5> of drying, it is preferable to dry at 40 to 50 o C The silver, and 42 to the still more preferably 48 ° C, most preferably 45 to 47 ° C. In addition, the drying is preferably dried for 5 to 10 days, more preferably for 6 to 9 days, most preferably for 7 to 8 days, in a specific embodiment of the present invention, the inventors Grind the wheat, mix water and infiltrate to form Nuruk, then ferment by inverting at 2 days intervals at 36 0 C and drying for 7 days at 45 0 C. The low acidity is effective in preventing microbial contamination, and the low amino acid level is expected to offset the taste felt during preparation of Takju (see Tables 2 and 5), and has excellent α-amylase and glucoamylase enzyme activity and glycosylation (Tables 3 and 6). ), The organic acid, free amino acid and fragrance components were analyzed to be excellent in taste and aroma quality (see Table 6, Table 8 and Table 10). By confirming that the remaining capacity of the enzyme is excellent and excellent storage (see Table 11 and Figure 8), the middle of the present invention fermented yeast and its manufacturing method can be usefully used as a manufacturing method of high quality Nuruk using the traditional method have. In addition, the present invention provides a yeast produced by the method for producing mesophilic fermentation medium of the present invention. In addition, the present invention provides a takju prepared by the yeast of the present invention.
상기 누룩은 효소활성 및 저장성이 우수한 것을 특징으로 한다. 본 발명의 구체적인 실시예에서， 본 발명의 중온 누룩제조 방법으로 제조된 누록이 미생물 오염 방지에 효과적이며 (표 2 및 도 5 참조)， 효소활성 및 당화력이 뛰어나고 (표 3 및 도 6 참조)， 맛과 향의 품질이 독특하고 우수할 것으로 ? 판단되며 (표 6， 표 8 및 표 10 참조)， 저장성이 우수함을 확인함으로써 (표 11 및 도 8 참조)， 본 발명의 중온 발효누룩 제조방법은 품질이 우수한 누룩 및 탁주 제조에 유용하게 이용될 수 있다. 이하， 본 발명을 실시예， 비교예 및 실험예에 의해 상세히 설명한다. The yeast is characterized by excellent enzyme activity and shelf life. In a specific embodiment of the present invention, the Nuruk produced by the method of manufacturing mesophilic yeast is effective for preventing microbial contamination (see Table 2 and FIG. 5), has excellent enzymatic activity and saccharification ability (see Tables 3 and 6), The taste and aroma quality will be unique and excellent ? Judgment (see Table 6, Table 8 and Table 10), by confirming the excellent storage properties (see Table 11 and Figure 8), the mesophilic fermented yeast production method of the present invention can be usefully used for the production of high-quality Nuruk and Takju Can be. Hereinafter, the present invention will be described in detail by Examples, Comparative Examples and Experimental Examples.
단ᅳ 하기 실시예， 비교예 및 실험예는 본 발명을 예시하는 것일 뿐， 본 발명의 내용이 하기 실시예， 비교예 및 실험예에 의하여 한정되는 것은 아니다. However, the following Examples, Comparative Examples and Experimental Examples are merely illustrative of the present invention, the content of the present invention is not limited to the following Examples, Comparative Examples and Experimental Examples.
<실시예 1>중온발효누룩제조 Example 1 Manufacture of medium temperature fermented yeast
전통방식을 이용하여 중온 발효한 누룩을 제조하여， 제조공정 기간 동안 누룩의 품질 변화를 확인하였다. A medium temperature fermented yeast was prepared using the traditional method, and the quality change of the yeast during the manufacturing process was confirmed.
구체적으로, 전남 광주지역에서 재배된 (2010년 산) 우리밀 (금강밀)을 구입하며， 누룩을 만들가 직전에 분쇄하여 사용하였다. 도 1에 나타난 바와 같이， 우리 밀을 를러 밀로 분쇄한 후, 7— mesh(2.8 mm) 채로 3번 반복하여 내려서， 가수량 26%로 살수 흔합하여 1시간 동안 실온에서 침윤시킨다. 침윤시킨 밀을 흔합한 다음, 340 g씩 무게를 달고 누룩 성형 를에 넣은 다음 성형장치를 이용하여 동일한 크기와 무게로 누룩을 성형한 후， 36°C 은도에서 2일 간격으로 뒤집기하며 발효시켰다. 건조장치를 이용하여 450C에서 7일 동안 건조시킨 후 발효기간으 3， 6， 10， 20 및Specifically, Woori wheat (Geumgang wheat) (grown in 2010), grown in Gwangju, Jeonnam, was purchased, and used before grinding to make yeast. As shown in Fig. 1, after crushing our wheat into a huller, it was lowered three times with 7-mesh (2.8 mm), and it was allowed to infiltrate at room temperature for 1 hour by spraying a mixture of 26% of hydrolysis. The infiltrated wheat was mixed, weighed 340 g each, and put into a yeast mold. The yeast was molded to the same size and weight by using a molding machine, and then fermented by inverting at 2 days intervals at 36 ° C. After drying for 7 days at 45 0 C using a drying apparatus, fermentation periods 3, 6, 10, 20 and
30일에 따른 품질특성의 분석 시료로 사용하였다 (도 1) . It was used as an analysis sample of quality characteristics according to 30 days (Fig. 1).
.  .
<비교예 1>자연 발효누룩 제조  Comparative Example 1 Preparation of Natural Fermented Nuruk
'상기 <실시예 1〉과 동일한 밀을 사용하여， 자연 발효시켜 누룩을 제조하여 대조군으로써 사용하였다. 'Using the same mill and the <Example 1> was used as a control group by natural fermentation to prepare a yeast.
구체적으로， 상기 <실시예 1>과 동일한 밀로 누룩을 만들기 직전에 를러 밀로 분쇄한 후， 7-mesh(2.8 mm) 채로 3번 반복하여 내려서， 가수량 26¾>로 살수 g 흔합하여 1시간 동안 실온에서 침윤시킨다. 침윤시킨 밀을 흔합한 다음 340 g씩 무게를 재서 성형장치를 이용하여 누룩을 성형한 후 20 내지 250C 온도에서 2일 간격으로 뒤집기하며 발효시켜 제조하였다. Specifically, immediately before making the yeast with the same mill as in <Example 1>, after grinding with a lorler mill, it is repeatedly lowered three times with 7-mesh (2.8 mm), sprayed with a hydrophilic amount 26¾> g Mix and infiltrate at room temperature for 1 hour. The infiltrated wheat was weighed and then weighed 340 g each to form a yeast using a molding apparatus, and then prepared by fermentation by inverting at intervals of 2 days at a temperature of 20 to 25 0 C.
<비교예 2>시판누룩 Comparative Example 2 Commercial Nuruk
상기 <실시예 1>에서 제조한 중온 발효누룩과 품질 비교를 위해， 현재 시판중인 부산의 산성누룩과 광주의 송학누룩을 구매하여， 동일하게 품질 분석 실험올 수행하였다 (도 2). 구체적으로， 산성누룩은 금정산성막걸리에서 구매하였고， 송학누룩은 송학곡자에서 구입하였다. For quality comparison with the mesophilic fermented yeast prepared in Example 1, a commercially available acidic yeast of Busan and Songhak nuruk of Gwangju were purchased, and the same quality analysis experiment was performed (FIG. 2). Specifically, acid yeast was purchased from Geumjeong Mt. Makgeolli, and Songhak yeast was purchased from Songhakgok.
<실험예 1>중은 발효누룩의 외형적 특징 규명 Experimental Example 1 Investigation of External Characteristics of Fermented Nuruk
상기 <실시예 1>에서 제조된 중온 발효누룩과 <바교예 2>의 시판누록 (산성누룩 및 송학누룩)의 외형적 변화를 비교하였다. The change in appearance of the medium-temperature fermented yeast prepared in <Example 1> and the commercial Nuruk (acidic Nuruk and Songhak Nuruk) of <Bagyo Example 2> were compared.
구체적으로 누룩의 무게는 Micro weighing scale (丽— Π 6000N, CAS Co.)로 측정하고 크기는 Digimatic c l iper(CD-20CPX, Mitutoyo Co.)로 측정하였으며， 누룩의 수분함량은 Moisture analyzer (MX— 50， AND Co.)로 측정하였다. Specifically, the weight of yeast was measured with a micro weighing scale (丽 — Π 6000N, CAS Co.) and the size was measured with a Digimatic cl iper (CD-20CPX, Mitutoyo Co.). 50, AND Co.).
그 결과 도 3에 나타난 바와 같이， 발효 0일 차에서는 중온 발효누룩과 시판누록의 외형적 차이는 크게 보이지 않았으며， 발효 초가에는 중온 발효누룩에서 당화능력을 가진 활털곰팡이 a/ ?e/7/a spp.)가 관찰되었다 (도 3). As a result, as shown in Figure 3, the day 0 of the fermentation did not show a significant difference between the medium-temperature fermented yeast and commercially available nucleus, and in the early stage of fermentation, the mildew mold with saccharifying ability in the medium-temperature fermented yeast a /? E / 7 / a spp.) was observed (FIG. 3).
또한， 누룩의 무게， 크기 및 수분함량을 분석한 결과 표 1에 나타난 바와 같이 누룩의 무게와 크기는 발효기간에 따라 전체적으로 감소하는 경향을 보였고， 이는 발효기간이 길어짐에 따라 발효과정 중 수분이 증발하기 때문인 것으로 생각되었다. 수분함량 분석 결과， 대조구인 산성누룩과 송학누룩은 11.9%와 8.66%였고， 중온 발효누룩의 수분함량은 1 내지 2%였다 (표 1). 보통 시판누룩의 경우 8 내지 10%의 수분함량을 보이나， 본 발명의 중온 발효누룩은 45°C에서 7일 동안 건조하는 g 과정을 거쳤기 때문에 전체적으로 수분함량이 1 내지 2%로 낮은 수치를 보였다. 본 발명에서 누룩의 건조과정을 인위적으로 거친 이유는 수분함량을 인위적으로 낮춤으로 해서 다른 미생물 생육을 저해하고， 누룩의 저장성을 높이기 위함이다. 【표 1】 In addition, as a result of analyzing the weight, size, and water content of yeast, as shown in Table 1, the weight and size of the yeast tended to decrease with the fermentation period. It was thought to be because. As a result of the water content analysis, the controls of acidic Nuruk and Songhak Nuruk were 11.9% and 8.66%, and the moisture content of medium-temperature fermented Nuruk was 1 to 2% (Table 1). In the case of commercial Nuruk, the water content of 8 to 10% is shown, but the medium-temperature fermented Nuruk of the present invention is dried at 45 ° C for 7 days. As a result of the g process, the total moisture content was low as 1 to 2%. The reason why artificially went through the drying process of nuruk in the present invention is to artificially lower the water content to inhibit the growth of other microorganisms and to increase the shelf life of the nuruk. Table 1
Figure imgf000011_0001
Figure imgf000011_0001
1〕값은 평균土 SD(N=3)이다. 1 ] value is mean 土 SD (N = 3).
<실험예 2>중은 발효누룩의 발효기간에 따른품온 변화확인 In Experimental Example 2, the change of product temperature according to the fermentation period of fermented yeast was confirmed.
상기 <실시예 1>에서 제조된 중온 발효누룩과 <비교예 1>의 자연 발효누룩의 품은 변화를 측정하였다.  The change of the medium-temperature fermented malt produced in <Example 1> and the natural fermented malt of <Comparative Example 1> were measured.
구체적으로， 발효 중 품온 변화는 H0B0®사의 data logger U12-013을 이용하여 발효기간 (0 3, 6 10 , 20 30일)에 따른 누룩의 품온 변화를 6시간 간격으로 측정하였다.  Specifically, the change in product temperature during fermentation was measured at 6 hour intervals using the H0B0® company's data logger U12-013 for changes in the leaven temperature during fermentation (0 3, 6 10, 20 30 days).
그 결과 도 4에 나타난 바와 같이 중온 발효누룩의 경우， 36°C에서 전발효를 진행하였으며， 0일부터 누룩미생물이 활동을 시작하여 2일째 가장 왕성한 번식을 하면서 최고 품은이 450C까지 올라갔 며 3일째부터 발효가 정체되면서 누룩의 품온이 360C로 발효 30일까지 일정하게 유지되었다 (도 4) . 온도와 품은 조절을 전혀 하지 않은 상태로 자연발효를 시킨 누룩의 경우， 20 내지 250C의 조건에서 발효를 진행하였고， 0일부터 누룩미생물이 활동을 시작하여 2일째 가장 왕성한 생육을 하면서 최고 품온이 27°C까지 증가하였고， 3일 이후부터는 24± 1°C로 품온이 일정하게 유지되었다. 이는 발효 온도 차이는 있지만 전체적으로 중온 발효누특과 동일한 미생물 생육과 善온 변화를 나타내었다. As a result, as shown in Fig. 4, in the case of medium-temperature fermented yeast, prefermentation was carried out at 36 ° C, and the yeast microorganisms started their activities from day 0, the most fertile breeding on the second day, and the highest boil rose to 45 0 C. As the fermentation stagnated from the 3rd day, The temperature was kept constant at 30 days of fermentation at 36 0 C (FIG. 4). In the case of leaven fermented by natural fermentation without any control of temperature and temperature, fermentation was carried out under the condition of 20 to 25 0 C. The leaven microorganisms started their activities from day 0 and grew the most vigorously on the second day. The temperature increased to 27 ° C, and after 3 days, the temperature was kept constant at 24 ± 1 ° C. It showed the same microbial growth and temperature change as the middle temperature fermentation algae, although there were differences in fermentation temperature.
<실험예 4>중은발효누룩의 이화학적 특성 변화확인 Experimental Example 4 Change of Physicochemical Properties of Middle Fermented Nuruk
상기 <실시예 1>에서 제조된 중은 발효누록의 발효 기간에 따른 이화학적 특성 변화를 pHᅳ 총산도 (Ac idi ty) 및 아미노산도 (Amino ac idi ty)를 측정하여 확인하였다. 구체적으로, 먼저 이화학적 특성 분석을 위한 누록 시료는 검사시료 20 g에 증류수 100 mL을 가하고 실온에서 3시간 침출하여 그 여과 용액을 준비하였다. 효소활성 분석을 위한 시료는 시료 10 g에 염화나트륨 용액 50 mL을 가해서 저은실 (4°C)에서 하룻밤 또는 실온 ( 15 내지 20°C)에서 3시간 동안 때때로 흔들어 침출한 후， 여과하여 실험의 조효소액으로 사용하였다. During the preparation in <Example 1>, the change in physicochemical properties of the fermented oak was determined by measuring pH ᅳ total acidity (Ac idi ty) and amino acid degree (Amino ac idi ty). Specifically, first, the Nuruk sample for physicochemical characterization was added 100 mL of distilled water to 20 g of the test sample and leached at room temperature for 3 hours to prepare the filtrate solution. Samples for enzyme activity analysis were added to 50 g of sodium chloride solution in 10 g of the sample, shaken occasionally at room temperature (4 ° C) overnight or at room temperature (15-20 ° C) for 3 hours, and then filtered. It was used as an enzyme solution.
pH는 pH meter (Or ion 3 star , Thermo scient i f ic Co . )로 실온에서 측정하였다. 총 산도는 일정량의 시료를 취하여 여과지로 여과한 검체 10 mL를 100 mL 삼각 플라스크에 취한 다음， 0.5% phenolphthalein 지시약을 2 내지 3방울 떨어뜨리고 0. 1 N NaOH로 적색이 될 때까지 중화 적정하였으며 소비된 용액의 양을 lact ic acid(%)로 환산하였다.  pH was measured at room temperature with a pH meter (Or ion 3 star, Thermo scienti Co Co.). The total acidity was determined by taking a certain amount of sample, taking 10 mL of the sample filtered with filter paper into a 100 mL Erlenmeyer flask, dropping 2-3 drops of 0.5% phenolphthalein indicator and neutralizing titration until it became red with 0.1 N NaOH. The amount of the prepared solution was converted into lactic acid (%).
아미노산도는 여과지로 여과한 검체 10 mL에 0.5% phenolphthalein 지시약을 2 내지 3방울 떨어뜨린 다음， 0. 1 N NaOH로 담흥색이 될 때까자 중화한 후， 여기에 중성포르말린 용액 5 mL을 가하여 유리된 산을 0. 1 N NaOH로 적정하여 담흥색이 될 때까지 소요된 0. 1 N NaOH의 mL수로 표시하였다. For amino acid, add 2 to 3 drops of 0.5% phenolphthalein indicator to 10 mL of the sample filtered with filter paper, neutralize it with 0.1 N NaOH until light gray, and then add 5 mL of neutral formalin solution. The acid was titrated with 0.1 N NaOH and expressed as the number of mL of 0.1 N NaOH consumed until light gray.
그 결과 하기 표 2 및 도 5에 나타난 바와 같이， 대조구로서 <비교예 2>의 n 시판누룩 중 산성누룩 추출물은 pH는 6.64였고， 송학누록의 추출물의 pH는 6.29였다. 본 발명의 중온 발효누룩의 경우， 발효기간에 따라 pH가 6. 18에서 5.84로 감소하였다 (표 2 및 도 5) . 시판 및 제조한 전통누룩의 추출물 pH는 약한 중성으로서 이들 간의 변화는 크지 않았다. 산도의 경우， 대조구인 산성누룩과 송학누룩의 산도는 0. 19%와 0.15%이었고， 중온 발효누룩은 발효기간에 따라 산도가 0.046%에서 0.078%로 증가하였다 (표 2 및 도 5) . 시판 누룩보다 제조한 중온 발효누룩의 산도가 2 내지 5배 낮았는데 이는 누룩 발효 초기에 위해 미생물 등의 잡균의 오염방지에 효과적일 것으로 예상되었다. 아미노산도의 경우， 산성누룩과 송학누룩은 1.28 mL로 같은 값이었고， 중온 발효누룩은 발효기간에 따라 아미노산도가 증가하다 발효 10일부터 발효기간에 따라 0.48 mL로 감소하였다 (표 2 및 도 5) . 따라서 제조한 중온 발효누룩의 아미노산도는 대조구로 사용한 시판 누룩보다 약 3배 정도 낮기 때문에 탁주 또는 약주를 제조하였을 때， 후미의 느끼한 맛이 많이 상쇄될 것이다. 이는 맛의 변화에 영향을 줄 것으로 판단된다. As a result, as shown in Table 2 and Figure 5, as a control of <Comparative Example 2> n The acidic malt extract of commercial Nuruk was 6.64, and the pH of the extract of Songhak Nuruk was 6.29. In the mesophilic fermented yeast of the present invention, the pH was decreased from 6. 18 to 5.84 according to the fermentation period (Table 2 and FIG. 5). Commercially produced and manufactured extracts of traditional yeast were weakly neutral, with little change between them. In case of acidity, the acidity of control acidic yeast and Songhak nuruk were 0.119% and 0.15%, respectively. In medium temperature fermented yeast, the acidity increased from 0.046% to 0.078% according to the fermentation period (Table 2 and Fig. 5). The acidity of the mesophilic fermented yeast produced was 2 to 5 times lower than that of commercial Nuruk, which was expected to be effective in preventing contamination of various microorganisms such as harmful microorganisms in the early stage of Nuruk fermentation. In the case of amino acidity, acidic Nuruk and Songhak Nuruk were the same value as 1.28 mL, and the intermediate temperature fermented Nuruk was increased in amino acidity with fermentation period. It decreased to 0.48 mL with fermentation period from 10 days of fermentation (Table 2 and Fig. 5). . Therefore, since the amino acidity of the produced medium-temperature fermented yeast is about three times lower than the commercial Nuruk used as a control, the taste of the aftertaste will be canceled much when Takju or Yakju is prepared. This may affect the change of taste.
【표 2】 아미노산도 [Table 2] Amino Acid Diagram
Λΐ- ττ:  Λΐ- ττ:
발효기간 (일) PH (0. 1N NaOH mL/10  Fermentation period (days) PH (0.1N NaOH mL / 10
(% , lact i c ac id)  (%, lact i c ac id)
mL) 대조구 (시판누룩) 산성 6.64±0.03 0. 1881±0.0068 1.28±0. 10 대조구 (시판 누룩) 송학 6.29±0.02 0. 1518+0.0143 1.28±0.01  mL) Control (commercial Nuruk) Acid 6.64 ± 0.03 0.01881 ± 0.0068 1.28 ± 0. 10 Control (Commercial Yeast) Transfer 6.29 ± 0.02 0.1518 + 0.0143 1.28 ± 0.01
"5"는 , " 5 " means,
≡. -H_ 0 6. 18±0.05 0.0462±0.0019 0.36±0.01 Iii. -H_ 0 6. 18 ± 0.05 0.0462 ± 0.0019 0.36 ± 0.01
1— Ξ 1— Ξ
^ τ  ^ τ
3 5.90±0.01 0.0708 ±0.0027 0.68+0.02 6 5.88±0.01 0.0786±으0018 0.70±0.02 3 5.90 ± 0.01 0.0708 ± 0.0027 0.68 + 0.02 6 5.88 ± 0.01 0.0786 ± 0018 0.70 ± 0.02
10 5.86±0.02 0.0735±0.0014 0.59±0.02 10 5.86 ± 0.02 0.0735 ± 0.0014 0.59 ± 0.02
20 5.85±0.01 0.0699±0.0014 0.48+0.0320 5.85 ± 0.01 0.0699 ± 0.0014 0.48 + 0.03
30 5.84±으01 0.0783 ±0.0009 0.48+0.03 값은 평균 ±SD(N=3)이다. 30 5.84 ± 01 0.0783 ± 0.0009 0.48 + 0.03 The mean value is ± SD (N = 3).
<실험예 3>중온 발효누룩의 효소학적 특성 변화확인 Experimental Example 3 Confirmation of Changes in Enzymatic Properties of Mesophilic Fermented Nuruk
상기 <실시예 1>에서 제조된 중온 발효누록의 발효 기간에 따른 효소학적 특성 변화를 a— amyl ase , glucoamylase , acidi c protease 활성 및 당화력 (SP)을 측정하여 확인하였다. The change in enzymatic properties according to the fermentation period of the mesophilic fermented oak prepared in <Example 1> was confirmed by measuring a— amyl ase, glucoamylase, acidic c protease activity and glycation potency (SP).
구체적으로， a-amylase 활성은 누룩 10 g에 염화나트륨 용액 50 mL를 가하여 저온에서 하룻밤 또는 실온에서 3시간 때때로 흔들어 침출한 후， 여과한 액을 희석하여 조효소액을 제조하였다. 1% 전분용액 2 mL을 시험관에 취해, 40°C에서 5분간 예열한 후， 효소액 0. 1 mL을 가해서 반웅을 개시하고， 반응액 중에서 0. 1 mL씩 피펫으로 1분 간격으로 미리 요오드용액을 10 mL을 넣어둔 시험관에 넣어 흔합하여 생성된 색을 25°C에서 유지하다가 두꺼운 10 mm을 통과하여 670 nm색을 비교하고, 투과율 1%을 측정하였다. 효소활성 (uni t )은 Wohlgemuth value에 준하여 하기 수학식 1로 산출하였다. Specifically, for a-amylase activity, 50 mL of sodium chloride solution was added to 10 g of yeast, and the solution was sometimes shaken at low temperature overnight or at room temperature for 3 hours, and then the filtrate was diluted to prepare a crude enzyme solution. Take 2 mL of 1% starch solution into the test tube, preheat it at 40 ° C for 5 minutes, add 0.1 mL of enzyme solution to start reaction, and pipette 0.01 mL of iodine solution into the reaction solution in 1 minute intervals. Was added to a 10 mL test tube to maintain the resulting mixture at 25 ° C. After passing through a thick 10 mm to compare the 670 nm color, 1% transmittance was measured. Enzyme activity (uni t) was calculated by the following Equation 1 based on the Wohlgemuth value.
[수학식 1][Equation 1]
Figure imgf000014_0001
Figure imgf000014_0001
Τ30 π1η ： 30분간 효소반웅을 시킨 후의 투과도 Τ 30 π 1η: Permeability after enzyme reaction for 30 minutes
T0 rain ： 효소반웅을 시키기 전의 투과도 Acidi c protease 활성은 누룩 사료 10 g에 염화나트륨 용액 50 mL를 가해서 저온실 (4°C)에서 하룻밤 또는 실온 ( 15 내지 20°C)에서 3시간 때때로 흔들어 침출한 후， 여과하였다. 상기 여액 10 mL를 투석막에 넣어 10 내지 2 M 초산 완충액에서 4°C 은도로 하룻밤 투석한 후， 상기 효소액을 희석하여 조효소액으로 하였다. 카제인 용액 1.5 mL에 pH 3.0 맥바인 완충액 1.0 mL와 조효소액 0.5 mL를 넣고 40oC에서 60분간 반응시킨 후， TCA용액 3 mL을 가해 반응을 정지시키고 침전을 제거하였다. 상기 액 1 mL에 탄산나트륨용액 5 mL와 페놀시약 1 을 가해서 40oC에서 30분간 발색시킨 후， 660 nm에서 흡광도를 측정하였고， 별도로 대조구로는 TCA용액을 가하기 직전에 효소액을 첨가하여 이하 위의 방법과 동일하게 하여 흡광도를 측정하여 하기 수학식 2에 의해서 효소활성을 산출하였다. [수학식 2]
Figure imgf000015_0001
T 0 rain : Permeability before enzyme reaction Acidi c protease activity was determined by adding 50 mL of sodium chloride solution to 10 g of yeast feed. After leaching overnight in a low temperature room (4 ° C.) or at room temperature (15-20 ° C.) for 3 hours, it was filtered. 10 mL of the filtrate was placed in a dialysis membrane and dialyzed overnight at 4 ° C. in 10 to 2 M acetic acid buffer, and the enzyme solution was diluted to form a crude enzyme solution. After adding 1.0 mL of pH 3.0 McVine buffer solution and 0.5 mL of coenzyme solution to 1.5 mL of casein solution, the mixture was reacted at 40 ° C. for 60 minutes, 3 mL of TCA solution was added to stop the reaction, and precipitation was removed. 5 mL of sodium carbonate solution and 1 phenol reagent 1 were added to 1 mL of the solution, followed by color development at 40 o C for 30 minutes, and the absorbance was measured at 660 nm. Separately, as a control, an enzyme solution was added immediately before adding the TCA solution. The absorbance was measured in the same manner as in the method, and the enzyme activity was calculated by the following Equation 2. [Equation 2]
Figure imgf000015_0001
*Glucoamylase 활성은 2% 전분용액 1 mL에 0.2 M 초산 완충액 0.2 mL를 가해서 40°C에서 5분간 예열한 후， 상기 누룩 효소액 0. 1 mL를 가해서 40oC에서 20분간 반웅시키고, 1. N NaOH용액 0. 1 mL을 첨가하여 반웅을 정지시킨 후, 30분간 방치하고 1 N 염산용액 0. 1 mL를 가해 중화하였다. 상기 실험에 사용한 대조구는 전분용액 1 mL에 0.2 M 초산 완충액 0.2 mL을 가해 40°C에서 5분간 예열하였다. 대조구는 시료와 동일하게 1 N 염산용액 0. 1 mL를 가해 중화시켰고, 효소활성은 DNS법으로 환원당을 측정하여 생성된 포도당량으로 하였다. Glucoamylase 활성은 가용성 전분으로부터 40oC에서 60분간 1 mg와 포도당을 생성하는 활성을 1 uni t로 하였으며， 시료 1 g의 glucoamylase 활성은 하기 수학식 3으로 산출하였다. * Glucoamylase activity was preheated for 5 minutes at 40 ° C by adding 0.2 mL of 0.2 M acetic acid buffer to 1 mL of 2% starch solution, and adding 0.1 mL of yeast enzyme solution and reacting for 20 minutes at 40 o C. The reaction was stopped by the addition of 0.01 mL of NaOH solution, left for 30 minutes, and neutralized by addition of 0.01 mL of 1 N hydrochloric acid solution. The control group used in the above experiment was preheated at 40 ° C. for 5 minutes by adding 0.2 mL of 0.2 M acetic acid buffer to 1 mL of starch solution. The control group was neutralized by adding 0.01 mL of 1 N hydrochloric acid solution in the same manner as the sample, and the enzyme activity was the amount of glucose produced by measuring the reducing sugar by DNS method. Glucoamylase activity was 1 uni t to produce 1 mg and glucose for 60 minutes at 40 o C from soluble starch, glucoamylase activity of the sample 1 g was calculated by the following equation (3).
[수학식 3] 효소 *성 t -생성포도당麵 반웅시 희석배쑤 당화력 (SP)을 측정을 위해, 먼저 상기 누룩 효소액을 희석배수 2개， 최소 10 mL씩 희석한 후， 조효소액으로 사용하였다. 2% 전분용액 50 mL과 식초산 완충용액 30 mL을 100 mL 용량 플라스크에 취한 다음 550C로 예열시킨 항온수조에서 10분 동안 예열시키고， 효소액 10 mL을 넣고 60분 동안 당화시켰다. 대조구는 증류수 10 mL을 넣고 60분 동안 동일하게 당화시켰다. 0. 5 N NaOH 10 mL을 넣고 효소반응을 정지시키고， 급넁시킨 후， 증류수를 가하여 100 mL로 조정하였다. [Formula 3] enzyme * sex t-produced glucose dilution In order to measure the glycation potency (SP), first, the yeast enzyme solution was diluted by 2 dilutions, at least 10 mL, and used as a coenzyme solution. 50 mL of 2% starch solution and 30 mL of vinegar acid buffer solution were taken in a 100 mL volumetric flask, preheated in a constant temperature water bath preheated to 55 0 C for 10 minutes, and 10 mL of enzyme solution was added and saccharified for 60 minutes. The control was added to 10 mL of distilled water and saccharified the same for 60 minutes. 10 mL of 0.5 N NaOH was added thereto, the enzyme reaction was stopped, and rapidly cooled, and then adjusted to 100 mL by adding distilled water.
당분 정량을 위하여 펠링 A시약 5 mL , 펠링 B시약 5 mL과 증류수 40 mL를 250 mL 삼각 플라스크에 넣고 준비한 후， 대조구로서 증류수 10 mL를 펠링시약 넣은 삼각플라스크에 넣고 삼각 플라스크를 끓여가며 표준 포도당 용액으로 적정하였다 (초반에 22 mL 정도 넣고 끓임. 24-26 mL정도 소비됨) . 펠링시약 내 황산구리의 청색이 점차 없어지면 메틸렌블루용액 4방울을 떨어뜨리고 용액을 계속 끓이면서 표준포도당 용액을 떨어뜨려 청색이 없어진 때를 종말점으로 하고， 이때 표준 포도당 용액의 소비 mL수를 B라 하였다. 다음 효소액 10 mL을 펠링시약올 넣은 삼각 플라스크에 넣은 후， 끓여가며 표준 포도당 용액으로 적정하는데, 초반에는 표준 포도당 용액을 넣지 않고 청색이 없어질 때쯤 메틸렌블루용액 4방울을 가하고 청색이 없어질 때까지 적정하였다. 적정을 완료하였을 때까지와 표준 포도당 용액의 소비 ^수를 M이라 하면 당화율은 하기 수학식 4 내지 6과 같이 산출하였다. To quantify the sugar, 5 mL of Pelling A reagent, 5 mL of Pelling B reagent and 40 mL of distilled water were prepared in a 250 mL Erlenmeyer flask, and then, as a control, 10 mL of distilled water was placed in an Erlenmeyer flask containing the Pelling reagent, and the Erlenmeyer flask was boiled. Titrate (beginning to 22 mL and boil, consume 24-26 mL). When the blue color of copper sulfate gradually disappeared, 4 drops of methylene blue solution was dropped, and the standard glucose solution was dropped by dropping the standard glucose solution while continuing to boil the solution. At this time, the number of mL of the standard glucose solution was defined as B. Next, add 10 mL of the enzyme solution into an Erlenmeyer flask containing Pelling reagent, boil and titrate with standard glucose solution.In the beginning, add 4 drops of methylene blue solution until blue color disappears without adding standard glucose solution until blue color disappears. Titration. When the titration was completed and the consumption ^ number of the standard glucose solution was M, the glycation rate was calculated as in Equations 4 to 6 below.
[수학식 4] 포도당 (mg)=2(B-M)x 100/효소반웅채취량 (mL) [수학식 5] 당화을 (»=누룩 1 g으로전 ·질 1 f t해하여생성 ¾S£당휼비 t로나타낸것 [수학식 6] [Equation 4] Glucose (mg) = 2 (BM) x 100 / Enzyme reaction volume (mL) [Equation 5] The glycosylation (»= 1g of yeast and vaginal 1 ft is produced to produce ¾S £ sugar ratio Burnt [Equation 6]
SP=당화율 x희석배수 그 결과 하기 표 3 맟 도 6에 나타난 바와 같이， α-amylase 활성은 대조구로서 <비교예 2>의 시판누룩 중 산성누룩 및 송학누룩은 175.1과 280.5 units/g이었다. 중온 발효누룩 (TN-A)은 발효기간에 따라 활성이 증가하였고， a-amylase activity는 310.3 units/g으로 시판 누룩보다 활성이 1.2 배 높았다 (표 3 및 도 6). Glucoamylase 활성은， 산성누룩과 송학누룩은 908과 3,372 units/g이었고， 발효기간에 따라 중온 발효누룩은 활성이 감소하다 증가하는 경향을 보였으며 , 5,592 units/g의 값으로， 대조구인 시판누룩보다 2 내지 3배 높은 활성을 보였다 (표 3 및 도 6). 결론적으로 <실시예 1>에서 제조한 중온 발효누룩의 당화력이 시판 누룩보다 높다는 것은 술을 빚을 때， 날곡류안누룩의 효소활성으로 쌀 전분을 당으로 분해시키는 효과가 크다는 것을 나타낸다.  As shown in Table 3 맟 Figure 6, the α-amylase activity was 175.1 and 280.5 units / g of commercial yeast of <Comparative Example 2> as a control. Medium-temperature fermented Nuruk (TN-A) increased its activity with fermentation period, and a-amylase activity was 310.3 units / g, 1.2 times higher than that of commercial Nuruk (Table 3 and Figure 6). The activity of glucoamylase was 908 and 3,372 units / g in acidic yeast and Songhak yeast, and the temperature of fermented yeast decreased with increasing fermentation period. The value of 5,592 units / g was higher than that of control. It showed 2-3 times higher activity (Table 3 and FIG. 6). In conclusion, the saccharification power of the mesophilic fermented yeast prepared in Example 1 is higher than that of commercial Nuruk, which indicates that the effect of decomposing rice starch into sugars by enzymatic activity of raw grain an Nuruk is great.
산성 단백질 활성은 산성누룩과 송학누록은 4,779.8과 4,212.5 units/g이었으며, 중온 발효누룩은 발효기간에 따라 활성아 증가하다 감소하는 경향을 보였고， 발효 30일차에 효소활성은 1,928.5 units/g으로 대조구인 시판 누룩보다는 3배정도 낮은 값을 보였다 (표 3 및 도 6). 특히， 누룩은 메주와 구성성분이 다른 관계로 지나칠 정도로 단백질 분해력이 높으면 단백질이 분해되어 아미노산으로 전환되는데 , 이때 주질의 맛 차이가 결정되기 때문에 단백질 분해력이 낮은 것이 유익하다.  The acidic protein activity was 4,779.8 and 4,212.5 units / g in acidic Nuruk and Songhak Yellow, and the temperature of fermented Nuruk increased with increasing fermentation time. The enzyme activity was 1,928.5 units / g at 30 days of fermentation. It was about 3 times lower than commercial leaven (Table 3 and FIG. 6). In particular, if the yeast is so proteolytically high enough that the meju and its components are different, the protein is broken down and converted into amino acids. At this time, since the taste difference of the main is determined, it is advantageous to have low proteolytic power.
당화력 (SP) 측정결과， 산성누룩과 송학누룩은 467.3과 915.5 SP이었고， 증온 발효누룩은 발효 3일째 당화력이 증가한 후， 그 이후에는 일정하게 유지되는 것을 볼 수 있었다. 발효 30일의 중온 발효누룩의 당화력은 1055.6 SP로 대조구인 시판 누룩보다 1.5배 우수한 것을 볼 수 있었다 (표 3 및 도 6). 당화력와 경우， 발효As a result of the measurement of glycation power (SP), the acidic yeast and Songhak yeast were 467.3 and 915.5 SP, and the temperature-fermented yeast was found to remain constant after the increase of saccharification on the third day of fermentation. The saccharification power of mesophilic fermented yeast at 30 days of fermentation was 1055.6 SP, which was 1.5 times better than that of the commercial yeast as a control (Table 3 and FIG. 6). Glycosylation and in cases
0일에도 719.33 SP 값을 보였는데 이는 450C에서 7일 동안 건조하는 과정 중에서 누룩의 발효가 진행된 것으로 생각되었다. 전체적으로 시판 누룩보다 본 발명의 중온 발효누록의 α-amylase, glucoamylase 활성과 당화력이 뛰어나고， 시판 누룩의 경우 산성단백질 분해력이 뛰어난 것을 알 수 있었다. On day 0, it showed 719.33 SP value, which was thought to be the fermentation of yeast during the drying process at 45 0 C for 7 days. Overall, it was found that α-amylase, glucoamylase activity and glycosylation activity of the mesophilic fermented nucleus of the present invention were superior to those of commercial Nuruk, and that the acidic protein decomposition ability of commercial Nuruk was excellent.
【표 3】  Table 3
Figure imgf000018_0001
Figure imgf000018_0001
<실험예 5>중은발효누룩의 생균수 변화확인 <Experiment 5> Confirmation of the viable cell number of fermented yeast
상기 <실시예 1〉에서 제조된 중온 발효누룩과 <비교예 2>의 시판누룩 (산성누룩 및 송학누룩)의 생균수 변화를 측정하였다.  The change in viable cell numbers of the medium-temperature fermented yeast prepared in <Example 1> and the commercial Nuruk (acidic Nuruk and Songhak Nuruk) of <Comparative Example 2> were measured.
구체적으로， 상기 누룩을 시료로 10 g을 취하여 멸균 백에 넣고 90 의 멸균 생리식염수를 가하며 균질기로 용해하고 멸균 생리식염수로 단계별로 희석한 후， PCA배지에 도말하고 나타난 균체수를 CFU/g로 표기하였다.  Specifically, take 10 g of the yeast as a sample, put it in a sterile bag, add 90 sterile saline solution, dissolve it with a homogenizer, dilute stepwise with sterile saline solution, smear the PCA medium, and display the number of cells in CFU / g. Notation.
그 결과 하기 표 4에 나타난 바와 같미， 대조구인 시판누록과 본 발명의 중은 발효누룩의 생균수는 109 내지 1010이었고， 발효기간에 따른 중온 발효누룩의 생균수는 발효 초기에 생육이 왕성하다가 발효기간이 길어짐에 따라 감소하는 경향을 볼 수 있었다 (표 4). 그리고 털 곰광이류 ( /c?ei?aw/a sp. , Rhizopus sp. , Mucor sp. 등)는 이 실험에서 제외를 하였다. As a result, as shown in Table 4 below, the control of commercially available Nuruk and the fermented yeast in the middle of the present invention was 10 9 to 10 10 , the viable count of mesophilic fermented yeast according to the fermentation period is vigorous growth at the beginning of fermentation As the fermentation period is longer, it tends to decrease (Table 4). And furry bearish (/ c? Ei? Aw / a sp., Rhizopus sp., Mucor sp. Etc.) were excluded from this experiment.
【표 4】  Table 4
Figure imgf000019_0001
Figure imgf000019_0001
값은 평균土 SD(N=3)이다.  The value is mean 土 SD (N = 3).
<실험예 6>중온 발효누룩의 유기산분석 Experimental Example 6 Organic Acid Analysis of Medium-Temperature Fermented Nuruk
상기 <실시예 1>에서 제조된 중온 발효누룩과 <비교예 2〉의 시판누룩 (산성누룩 및 송학누룩)와유기산을 분석하였다. The medium-temperature fermented yeast prepared in <Example 1> and commercial Nuruk (acidic Nuruk and Songhak Nuruk) and organic acid of <Comparative Example 2> were analyzed.
구체적으로, 누룩 시료 1 g에 20% ethanol 5 tnL을 가한 후， 30°C 은도에서 ?0분 동안 sonicator에서 초음파하면서 추출하고, 원심분리기에서 3,000 rpm, 4°C, 10분 동안 원심여과한 후， 20% ethanol 5 mL을 가하고 위와 같은 방법으로 3희 반복하여 총 20 mL이 되게 하였다. 상등액을 취하여 0.2 /Λΐι membrane filter (Mullepore Cp. , Ireland)로 여과한 후， 분석하였다. 유기산 분석은 하기 표 5의 조건으로 HPLC X-20A Prominence, Shimadzu Co., Japan)를 사용하였으며， column은 KO 811(7.8x300 mm, Shodex Co. ) , .mobile phase는. 4 mM sulfuric acid를 ■사용하였으며 flow rate는 0.6 mL/min, inject ion volume은 10 detector는 PDA(210 run) 사용하였다 (표 5). 【표 5】 Specifically, after adding 20% ethanol 5 tnL to 1 g of the yeast sample, extracted by sonicator for 0 minutes at 30 ° C silver, and centrifuged at 3,000 rpm, 4 ° C, 10 minutes in a centrifuge ， 5 mL of 20% ethanol was added and repeated 3 times in the same manner to make a total of 20 mL. The supernatant was taken and filtered with a 0.2 / Λΐι membrane filter (Mullepore Cp., Ireland) and analyzed. For organic acid analysis, HPLC X-20A Prominence, Shimadzu Co., Japan) was used under the conditions of Table 5, and the column was KO 811 (7.8 × 300 mm, Shodex Co.), .mobile phase. 4 mM sulfuric acid ■ The flow rate was 0.6 mL / min and the inject ion volume was 10 detectors using PDA (210 run) (Table 5). Table 5
Figure imgf000020_0001
그 결과 하기 표 6에 나타난 바와 같이， 대조구로서 <비교예 2>의 시판 누룩인 산성누룩의 경우 citric acid (구연산)가 주요 유기산으로 검출되었고， 송학누룩은 citric acid, malic acid (사과산)와 lactic acid (젖산)가 주요 유기산으로 분석되었다 (표 6). 본 발명의 <실시예 1>에서 제조한 중온 발효누룩의 경우, citric acid, malic acid, oxalic acid가 분석되었으며， 주요 유기산으로 citric acid (구연산)은 발효 초기에 증가하였다가 10일부터 일정하게 유지되었다 (표 6). 누룩별로 유기산의 양과 종류는 차이가 있었는데 이는 원재료, 관여 발효미생물 및 발효조건에 따른 차이로 생각되었다.
Figure imgf000020_0001
As a result, as shown in Table 6, citric acid (citric acid) was detected as a major organic acid in the acidic malt which is a commercial malt of <Comparative Example 2>, and Songhak malt was citric acid, malic acid (apple acid) and lactic. acid (lactic acid) was analyzed as the main organic acid (Table 6). In the mesophilic fermented yeast prepared in <Example 1> of the present invention, citric acid, malic acid, and oxalic acid were analyzed, and citric acid (citric acid) increased as early organic fermentation and kept constant from 10 days (Table 6). The amount and type of organic acid differed by yeast, which was considered to be different according to raw materials, related fermentation microorganisms and fermentation conditions.
【표 6] [Table 6]
구성 (mg%) Composition (m g %)
대조구 대조구 중온 발효누룩 (days)  Control Control Medium Temperature Fermented Nuruk (days)
유기산  Organic acid
산성누 송학누  Acid Silk
0 3 6 10 20 30  0 3 6 10 20 30
2.3.43士 2.63土 10.49土2.3.43 士 2.63 土 10.49 土
Oxalic n.d2) n.d n.d n.d n.d Oxalic nd 2 ) nd nd nd nd
5.09 1.05 0.99 298.52 5.09 1.05 0.99 298.52
186.65 145.89 205.95 189.35 174.83 164.66 155.56 186.65 145.89 205.95 189.35 174.83 164.66 155.56
Citric ±45.891 Citric ± 45.89 1
±71.53 +22.03 土 29.96 ±29.23 ±4.32 ±2.66 ±76.64 )  ± 71.53 +22.03 土 29.96 ± 29.23 ± 4.32 ± 2.66 ± 76.64)
Tartar 2.03± 1 2.57士  Tartar 2.03 ± 1 2.57 士
n.d n.d n.d n.d n.d n.d ic .05 0.23  n.d n.d n.d n.d n.d n.d ic .05 0.23
28.78  28.78
57.23士 41.04士 36.19士 35.09+ 29.65 22.20士 57.23 士 41.04 士 36.19 士 35.09+ 29.65 22.20 士
Malic n.d ± 12.7 Malic n.d ± 12.7
19.74 4.21 15.26 1.05 ±6.40 1.05  19.74 4.21 15.26 1.05 ± 6.40 1.05
2  2
Succin  Succin
n.d n.d n.d n.d n.d n.d n.d n.d ic  n.d n.d n.d n.d n.d n.d n.d n.d ic
Fumari  Fumari
n.d n.d n.d n.d . , n.d n.d n.d n.d c  n.d n.d n.d n.d. , n.d n.d n.d n.d c
33.28 +  33.28 +
Lactic n.d n.d n.d n.d n.d n.d n.d  Lactic n.d n.d n.d n.d n.d n.d n.d
11.37  11.37
Formi  Formi
n.d n.d n.d n.d n.d n.d n.d n.d c  n.d n.d n.d n.d n.d n.d n.d n.d c
Acetic n.d n.d n.d n.d n.d n.d n.d n.d Acetic n.d n.d n.d n.d n.d n.d n.d n.d
Pyrogl Pyrogl
n.d n.d n.d n.d n.d n.d n.d n.d utamic  n.d n.d n.d n.d n.d n.d n.d n.d utamic
Total 298.52 277.16 210.36 244.17 224.44 204.49 198.65 188.25  Total 298.52 277.16 210.36 244.17 224.44 204.49 198.65 188.25
υ값은 평균 ±SD(N=3)이다. 2)n. d는 검출되지 않았다. The value υ is the mean ± SD (N = 3). 2) n. d was not detected.
<실험예 7>중온 발효누룩의 발효기간에 따른 밀누룩의 유리 아미노산분석 상기 <실시예 1>에서 제조된 중온 발효누룩과 <비교예 2>의 시판 누룩 (산성누룩 및 송학누룩)의 유리 아미노산을 분석하였다. 시료 1 g에 80% ethanol 25 mL를 넣고 homogeni zer로 균질화한 후， 30°C에서 15분 추출하였다. 추출물을 4°C에서 3 , 000 rpm으로 10분 동안 원심분리하여 상등액을 1 mL 또는 2 mL 취하여 농축기 또는 dry oven을 사용하여 증발 건조시킨 다음， 0.02 N HC1 1 mL 또는 2 mL를 가하여 잘 녹이고， 0.2 Syr inge f i I ter (Whatman Co . , USA)로 여과한 후'， amino acid analysis system(L— 8900, Hitachi Co. , Japan)을 이용하여 하기 표고조건으로 분석하였다. Experimental Example 7 Analysis of Free Amino Acids of Wheat Nuruk According to Fermentation Period of Mesophilic Fermented Nuruks The free amino acids of mesophilic fermented yeast prepared in <Example 1> and commercially produced Nuruk (acidic Nuruk and Songhak Nuruk) of <Comparative Example 2> Was analyzed. 25 g of 80% ethanol was added to 1 g of the sample, homogenized with a homogeni zer, and extracted at 30 ° C. for 15 minutes. Centrifuge the extract at 4 ° C for 3, 000 rpm for 10 minutes, take 1 mL or 2 mL of the supernatant, evaporate to dryness using a concentrator or dry oven, dissolve well by adding 1 mL or 2 mL of 0.02 N HC1. 0.2 Syr inge fi I ter (Whatman Co., USA) and then filtered using ' , amino acid analysis system (L-8900, Hitachi Co., Japan) under the following elevation conditions.
[표 7】  TABLE 7
Figure imgf000022_0001
그 결과 하기 표 8에 나타난 바와 같이， <비교예 2>의 시판중인 산성누룩의주요 아미노산은 proline (단맛 /쓴맛), alanine (단맛)， valine (약한 쓴맛)， glutamic acid (감칠맛)， γ-aminobutyric acid 등 14개 유리 아미노산으로 구성되었고， 총 아미노산의 함량은 602.92 /g/mL이었으며， 이 중 필수 아미노산의 .함량은 21.14 /g/mL이었다. 송학누룩은 주요 유리 아미노산은 proline (단맛 /쓴맛)， alanine (단맛)， valine (약한 쓴맛)， urea, α-aminobutyr ic acid 등 24개 유리 아미노산으로 구성되었고， 총 아미노산의 함량은 929.98 /g/mL아었고， 이 중 필수 아미노산의 함량은 103.63 /g/m£이었다 (표 8) .
Figure imgf000022_0001
As a result, as shown in Table 8, the main amino acids of commercial yeast in Comparative Example 2 were proline (sweet / bitter), alanine (sweet), valine (light bitter), glutamic acid (mild), γ- It was composed of 14 free amino acids such as aminobutyric acid, and the total amino acid content was 602.92 / g / mL, and the essential amino acid content was 21.14 / g / mL. The major free amino acids of Songhak Nuruk consisted of 24 free amino acids including proline (sweet / bitter), alanine (sweet), valine (weak bitter), urea, α-aminobutyr ic acid, and the total amino acid content is 929.98 / g / mL, which is required The amino acid content was 103.63 / g / m £ (Table 8).
본 발명의 중온 발효누룩의 발효 30일차 주요 유리 아미노산은 prol ine (단맛 /쓴맛)， val ine (약한 쓴맛)， taur ine , alanine , γ-aminobutyr ic acid 등 ' 8개 융리 아미노산으로 구성되었고， 총 아미노산의 함량은 107.23 g/mL이었고， 이 중 팔수 아미노산의 함량은 12.27 / g/mL이었다 (표 8) . Fermentation 30 primary key amino acids of mesophilic fermenting yeast of the invention prol ine (sweet / bitter taste), val ine (weak bitter taste), taur ine, alanine, γ-aminobutyr ic acid such as' was composed of eight yungri amino acids, a total of The amino acid content was 107.23 g / mL, of which the amino acid content was 12.27 / g / mL (Table 8).
유리 아미노산 분석 결과, 중온 발효누룩보다 시판 누룩의 유리 아미노산의 함량이 높고 다양한 아미노산으로 구성되어 있는 것을 볼 수 있었다. 이는 원재료, 제조 지역의 기후 환경 및 발효조건의 차이로 생각되었고， 이 발효제로 탁주를 제조하였을 때 이들의 주질 특성은 뚜렷할 것으로 추측되었다. As a result of free amino acid analysis, it was found that the content of free amino acid of commercial Nuruk was higher than that of medium-temperature fermented Nuruk and composed of various amino acids. This was considered to be a difference between the raw materials, the climatic environment of the manufacturing region and the fermentation conditions, and it was assumed that the main characteristics of Takju were obvious when the fermentation agent was prepared.
【표 8]  [Table 8]
유리 대조구 중온 발효누룩 /g/mL, day)  Glass control medium fermented yeast / g / mL, day)
아미노 Amino
산성누룩 .송학누룩 0 3 6 10 20 30 산 Acid Yeast . Songhak Yeast 0 3 6 10 20 30 Mountain
'8.71 ±0.6 16.49士 1. ' 8.71 ± 0.6 16.49 士 1.
P-Ser n.d n.d n.d n.d n.d n.d  P-Ser n.d n.d n.d n.d n.d n.d
51) 05 5 1) 05
10.47± 1, 6.67±0. 6.25± L 5.04±2. 9.78± 1. 6.17士2. 10.47 ± 1, 6.67 ± 0. 6.25 ± L 5.04 ± 2. 9.78 ± 1. 6.17 士 2.
Tau n.d¾ n.d Tau n.d¾ n.d
.55 38 68 26 48 65 .55 38 68 26 48 65
18·49±3. 34.41 ±4,. 18.49 ± 3. 34.41 ± 4,.
Urea n.d n.d n.d n.d n.d n.d  Urea n.d n.d n.d n.d n.d n.d
84 77  84 77
10.03士 1. 그 67±4.0  10.03 士 1.That 67 ± 4.0
Asp n.d n.d n.d n.d n.d n.d  Asp n.d n.d n.d n.d n.d n.d
72 5  72 5
1.58土 0.1 8.11±4.9 ·  1.58 土 0.1 8.11 ± 4.9
Thrc n.d n.d n.d n.d n.d n.d  Thrc n.d n.d n.d n.d n.d n.d
2 1  2 in 1
2.53士 0.7 9.29±5.8  2.53 士 0.7 9.29 ± 5.8
Ser n.d n.d n.d n.d n.d n.d  Ser n.d n.d n.d n.d n.d n.d
3 9  3 9
20.43±3. 15.78± 1.  20.43 ± 3. 15.78 ± 1.
Glu n.d n.d n.d n.d n.d n.d  Glu n.d n.d n.d n.d n.d n.d
64 13  64 13
13.38±2. 9.30±0.7 3.00±0. 2.26± 1. 2.50±0. 3.81± 1. 2.04±0. 1.44±0. 13.38 ± 2. 9.30 ± 0.7 3.00 ± 0. 2.26 ± 1.2.50 ± 0. 3.81 ± 1.2.04 ± 0. 1.44 ± 0.
Gly Gly
52 4 40 01 84 05 53 10 52 4 40 01 84 05 53 10
36.57±8. 44.02士 2. 2.47±0. 2.68±0. 2.98± 1. 4.25±0. 2.57±0. 3.35±0.36.57 ± 8. 44.02 士 2. 2.47 ± 0. 2.68 ± 0. 2.98 ± 1. 4.25 ± 0. 2.57 ± 0. 3.35 ± 0.
Ala Ala
47 61 66 51 . 57 87 80 32 47 61 66 51. 57 87 80 32
1.70± 1.0 1.70 ± 1.0
Cit n..d n.d n.d n.d .6.77 . n.d n.d  Cit n..d n.d n.d n.d .6.77. n.d n.d
5  5
.28.84士4. 15.91土 3 . 28.84 士 4. 15.91 土 3
a- ABA n.d n.d n.d n.d n.d n.d a- ABA n.d n.d n.d n.d n.d n.d
85 .52 22.56士 0. 38.66士2. 12.38士 0 13.80±4 12.14±2 8.74±4. 13.35士 4 12.27土 785 .52 22.56 士 0.38.66 士 2. 12.38 士 0 13.80 ± 4 12.14 ± 2 8.74 ± 4. 13.35 士 4 12.27 土 7
Vale Vale
81 15 .58 .74 .35 32 .75 .89 81 15 .58 .74 .35 32 .75 .89
4.09± 1.0 7.67±.0. 10.01± 1 10.58±0 8.94土 L 4.09 ± 1.0 7.67 ± .0. 10.01 ± 1 10.58 ± 0 8.94 土 L
Cys n.d n.d n.d  Cys n.d n.d n.d
5 40 .05 .92 05  5 40 .05 .92 05
22.47土 5.  22.47 土 5.
Ilec n.d n.d . n.d n.d n.d n.d n.d Ile c nd nd . nd nd nd nd nd
65  65
19.09±2.  19.09 ± 2.
Leuc n.d n.d n.d n.d n.d n.d n.d Leu c nd nd nd nd nd nd nd
95  95
21.64士0.  21.64 士 0.
Phe n.d n.d n.d n.d n.d n.d n.d  Phe n.d n.d n.d n.d n.d n.d n.d
39  39
17. Γ7±3. 4.89士0.5 17.Γ7 ± 3 . 4.89 士 0.5
b-Ala n.d n.d n.d n.d n.d n.d b-Ala n.d n.d n.d n.d n.d n.d
04 5  04 5
b一 1.64±0.1  b 一 1.64 ± 0.1
n.d n.d n.d n.d n.d n.d n.d AiBA 5  n.d n.d n.d n.d n.d n.d n.d AiBA 5
19.80±4. 16.45±9. 4.00±0. 1.53士ᄋ. 1.38±0. 1.72±0'. 3.07土 0. 2.01 ± 1. g-ABA  19.80 ± 4. 16.45 ± 9. 4.00 ± 0. 1.53 士 ᄋ. 1.38 ± 0. 1.72 ± 0 '. 3.07 土 0. 2.01 ± 1. g-ABA
32 16 93 20 54 28 72 05 . 32 16 93 20 54 28 72 05.
68.89±3. 62.20±4. 71.02±5 42.14±6 21.65土 3 31.29±2 19.70± 1 36.36±368.89 ± 3. 62.20 ± 4. 71.02 ± 5 42.14 ± 6 21.65 土 3 31.29 ± 2 19.70 ± 1 36.36 ± 3
NH3 NH3
53 16 .93 .76 . .90 .17 .59 .08 53 16 .93 .76. .90 .17 .59 .08
2.13土 0.1 2.13 土 0.1
Lysc n.d n.d n.d n.d n.d n.d n.d Lys c nd nd nd nd nd nd nd
5  5
5.63±0.1  5.63 ± 0.1
Hisc n.d n.d n.d n.d n.d n.d n.d His c nd nd nd nd nd nd nd
5  5
6.36±0.6  6.36 ± 0.6
Argc n.d n.d n.d n.d n.d n.d n.d  Argc n.d n.d n.d n.d n.d n.d n.d
7  7
2.10± 1.0 2.04± 1.2 2.40± 1. 1.10±0. 4.17± 1. 1.54士 1. 2.10 ± 1.0 2.04 ± 1.2 2.40 ± 1. 1.10 ± 0. 4.17 ± 1.1.54 士 1.
Hypro n.d n.d Hypro n.d n.d
5 6 05 15 28 05 5 6 05 15 28 05
360.69士 547.07士 1.18±0. 39.72± 1 43.54±5 49.53± 1 Γ7.15±0 44·08±8360.69 士 547.07 士 1.18 ± 0. 39.72 ± 1 43.54 ± 5 49.53 ± 1 Γ7.15 ± 0 44 · 08 ± 8
Pro Pro
9.49 4.83 78 .05 . .23 .58 .91 .16 9.49 4.83 78 .05. .23 .58 .91 .16
Totals 602.92 929.98 128.10 118.82 103.41 127.15 71.83 107.23 필수 Totals 602.92 929.98 128.10 118.82 103.41 127.15 71.83 107.23 Required
아미노 24.14 103.63 12.38 13.80 12.14 8.7 13.35 12.27 산 3> Amino 24.14 103.63 12.38 13.80 12.14 8.7 13.35 12.27 acid 3 >
값은 평균土 SD(N=3)이다. The value is mean 土 SD (N = 3).
2) n. d는 검출되지 않았다. 2) n. d was not detected.
3) 필 아미노산 <실험예 8>중은발효누룩의 향기성분분석 상기 <실시예 1>에서 제조된 중온 발효누특과 <비교예 2>의 시판 누룩 (산성누룩 및 송학누룩)의 향기성분을 분석하였다. 구체적으로, 누룩 시료 10 g에 에테르 -펜탄 흔합용액 (1:1) 100 mL을 넣고 150 rpm 20oC에서 하룻밤 동안 추출한 후， 여과하고 걸러진 시료에 에테르 -펜탄 흔합용액 (1:1) 50 mL을 넣고 150 rpm, 20oC에서 1시간 동안 재 추출하였다. 두 추출용액을 흔합하여 sodium sulfate를 첨가하여 물이 전부 흡수될 때까지 흔들어준 다음， 이 액을 여과하고 농축한 후， 0.2 \Ά Syringe filter (Whatman Co. USA)로 여과하여 GCMS (GCMS-QP2010plus)을 이용하여 하기 표 9의 조건으로 분석하였다 (표 9). 3) Peel Amino Acid <Experiment 8> Analysis of Flavor Components in Fermented Nuruk The fragrance components of the medium-temperature fermented silkworms prepared in <Example 1> and the commercial Nuruk (acidic Nuruk and Songhak Nuruk) of <Comparative Example 2> were analyzed. Specifically, 100 mL of the ether-pentane mixed solution (1: 1) was added to 10 g of the yeast sample, extracted at 150 rpm 20 o C overnight, and then filtered and the filtered sample 50 mL of the ether-pentane mixed solution (1: 1). And re-extracted at 150 rpm, 20 o C for 1 hour. Combine the two extraction solutions, add sodium sulfate, shake until all the water is absorbed, filter the solution, concentrate it, filter with 0.2 \ Sy Syringe filter (Whatman Co. USA), and GCMS (GCMS-QP2010plus). ) Was analyzed under the conditions of Table 9 below (Table 9).
【표 9】 내용 조건 [Table 9] Contents condition
GC-MS QP2010plus GC-MS QP2010plus
HP-INN0WAX (60 m χθ.25 mm i,d χθ.25 pm film thikness) 주입 온도 250°C 컬럼 속도 1.0 cm/sec (linear velocity, N2) 분리량 HP-INN0WAX (60 m χ θ.25 mm i, d χ θ.25 pm film thikness) Injection temperature 250 ° C Column velocity 1.0 cm / sec (linear velocity, N2)
20  20
(Split ration)  (Split ration)
23013 23013
30 m in  30 m in
5 요 ψ  5 yo ψ
2 min 2 min
FID 250°C 주입 부피 1 id FID 250 ° C injection volume 1 id
그 결과 하기 표 10에ᅵ 나타난 바와 같이 hydrocarbone 계열의 32종， ester 계열 0 4 32 species of hydrocarbone series, as a result appears to i are shown in Table 10, ester series 0 4
10종， acid 계열이 4종， alco )l류가 2종으로 총 48종의 향기성분이 검출되었다. 분석한 3종의 (중온 발효누룩， 산성누룩 및 송학누룩) 누룩의 공통 향기성분은 octane, formic acid ethyl ester , ( E ) -6-me t hy 1 -3-undecene , decane, methyl benzene , undecane, metyhl laurate, 1,3ᅳ dimethyl—benzene, 1 , 2-d i methyl -benzene , l_ethyl_3ᅳ methylᅳ benzene, acidic acid, n-hexadecanoin acid의 12개 성분으로 구성되어져 있었다 (표 10). A total of 48 aroma components were detected, including 10 species, 4 types of acid series, and 2 types of alco) l. The common fragrance components of the three types of yeast (medium-fermented yeast, acidic yeast and Songhak yeast) were octane, formic acid ethyl ester, (E) -6-met hy 1-3-undecene, decane, methyl benzene and undecane It was composed of 12 components, metyhl laurate, 1,3 'dimethyl-benzene, 1, 2-dimethyl-benzene, l_ethyl_3' methyl ᅳ benzene, acidic acid and n-hexadecanoin acid (Table 10).
누룩별 주요성분은 산성누룩은 (9Z,12Z)-9, 12-octadecadienoate ethyl , ethyl palmitate, n-hexadecanoic acid, 8-heptadecene, 9-octadecenoic acid 등 종 22개' 성분으로, 송학누룩은 acetic acid ethyl ester , n-hexadecanoic acid, octadec-9- enoic acid, methyl octadeca-9, 12-dienoate, 1-tr icosene 등 .27개 '성분，■ 중은 발효누룩 (TN_A)는 acetic acid ethyl ester , n-hexadecanoic acid, 4-ethoxy- benzoic acid ethyl ester , 1 , 3-d i me t hy 1 -benzene , 2-methyl -octane 등ᅵ종' 25개 성분으로 분석되었다 (표 10). 특히. 시판누룩과 본 발명의 증온 발효누룩과의 향기의 구성성분은 큰 차이는 보이지 않는다. Key Ingredients yeast is the yeast acid (9Z, 12Z) -9, 12 -octadecadienoate ethyl, ethyl palmitate, n-hexadecanoic acid, 8-heptadecene, 9-octadecenoic acid species such as 22 to "ingredient songhak yeast is acetic acid .27 ' components such as ethyl ester, n-hexadecanoic acid, octadec-9-enoic acid, methyl octadeca-9, 12-dienoate, 1-tr icosene, etc. ■ Medium fermented yeast (TN_A) is acetic acid ethyl ester, n -hexadecanoic acid, 4-ethoxy- benzoic acid ethyl ester, 1, 3-di me t hy 1 -benzene, 2-methyl -octane such species i, was analyzed by 25 component (Table 10). Especially. The composition of the fragrance between commercial Nuruk and the elevated temperature fermented Nuruk of the present invention does not show a big difference.
【표 10】  Table 10
RT RT
피크 서부 주오 Peak West Chuo
(min) 사_서ᄋ누 1 ^ 송학누룩  4 mins 1
발효누룩  Fermented Yeast
1 5.467 Octane 0.72 . 0.81 1.03  1 5.467 Octane 0.72. 0.81 1.03
2 5.861 3ᅳ Methyl— octane 0.29  2 5.861 3 ᅳ Methyl— octane 0.29
3 5.982 Formic acid ethyl ester 0.48 0.6 0.66  3 5.982 Formic acid ethyl ester 0.48 0.6 0.66
4 6.445 Dodecane 1.12 .  4 6.445 Dodecane 1.12.
5 6.448 2-Methyl-octane 1.45  5 6.448 2-Methyl-octane 1.45
6 6.574 (E)-6-Methyl-3-undecene 0.57 0.63 0.75  6 6.574 (E) -6-Methyl-3-undecene 0.57 0.63 0.75
7 6.793 Tetrahydro-2-methyl-furan 0.25  7 6.793 Tetrahydro-2-methyl-furan 0.25
8 7.344 Nonane 0.6 0.86  8 7.344 Nonane 0.6 0.86
9 9.151 2-Methyl-nonane . 0.35  9 9.151 2-Methyl-nonane. 0.35
10 10.76 Decane 0.34 0.4 0.69 ' 10 10.76 Decane 0.34 0.4 0.69 ''
11 11.322 3,3-Dimethyl-octane 0.28. 11 11.322 3,3-Dimethyl-octane 0.28.
12 12.574 3，그 Dimethyl decane 0.39  12 12.574 3 ， Dimethyl decane 0.39
13 12.805 Methyl benzene 0.93 ' 0.98 1.35 14 15.876 Undecane 0.17 0.2 0.3313 12.805 Methyl benzene 0.93 ' 0.98 1.35 14 15.876 Undecane 0.17 0.2 0.33
15 17.57 Methyl laurate 0.35 0.39 15 17.57 Methyl laurate 0.35 0.39
16 17.593 Ethyl-benzene . 0.51  16 17.593 Ethyl-benzene. 0.51
17 18.467 1,3— Dim6thy卜 benzene 1.07 1.17 1.65  17 18.467 1,3— Dim6thy 卜 benzene 1.07 1.17 1.65
18 21.236 1，2 Dimethyl benzene 0.48  18 21.236 1,2 Dimethyl benzene 0.48
19 21.249 Xylene 0.5 0.69 19 21.249 Xylene 0.5 0.69
■ 20 23.861 1 Ethyl 3 methyl— benzene 0.52 0.59 0.86 ■ 20 23.861 1 Ethyl 3 methyl— benzene 0.52 0.59 0.86
21 25.12 1，3，5 Trimethyl benzene 0.21  21 25.12 1,3,5 Trimethyl benzene 0.21
22 27.637 1,2,4-Trimethyl-benzene 0.78  22 27.637 1,2,4-Trimethyl-benzene 0.78
23 27.649 1，3，5 Trimethyl— benzene 0.85 1.28  23 27.649 1，3，5 Trimethyl— benzene 0.85 1.28
24 38.34 3-Hexadecene 0.6  24 38.34 3-Hexadecene 0.6
25 39.584 Acetic acid ： 0.43 0.69 1.07  25 39.584 Acetic acid ： 0.43 0.69 1.07
26 44.157 Acetic acid ethyl ester 23.55 33.8  26 44.157 Acetic acid ethyl ester 23.55 33.8
27 45.258 2,3-Butanediol 0.61  27 45.258 2,3-Butanediol 0.61
28 47.947 n-Pentadecane . 0.46 . 0.31 28 47.947 n-Pentadecane . 0.46. 0.31
29 50.651 1-Hexadecene 0.53 2.45  29 50.651 1-Hexadecene 0.53 2.45
30 50.671 9-Octadecene 0.95  30 50.671 9-Octadecene 0.95
31 61.891 9-Eicosene 1.0  31 61.891 9-Eicosene 1.0
32 61.909 1-Octadecene 2.97  32 61.909 1-Octadecene 2.97
33 66.855 8-Pentadecanone 0.52  33 66.855 8-Pentadecanone 0.52
34 72.137 3-Eicosene 0.84  34 72.137 3-Eicosene 0.84
35 72.14 1-Nonadecene  35 72.14 1-Nonadecene
36 72.314 Ethyl tetradecanoate 0.56  36 72.314 Ethyl tetradecanoate 0.56
37 78.364 4-Ethoxy benzoic acid ethyl ester 1.7  37 78.364 4-Ethoxy benzoic acid ethyl ester 1.7
38 81.908 Ethyl palmitate 7.7  38 81.908 Ethyl palmitate 7.7
39 90.247 1-Tricosene 4.3- 39 90.247 1-Tricosene 4.3-
. 40 90.694 Methyl octadeca 9,12 dienoate 7.99 . 40 90.694 Methyl octadeca 9,12 dienoate 7.99
41 90.745 Ethyl stearate 1.03  41 90.745 Ethyl stearate 1.03
42 91.598 9-Octadecenoic acid ethyl ester 5.37  42 91.598 9-Octadecenoic acid ethyl ester 5.37
(9Z, 12Z)-9, 12-Octadecadienoate  (9Z, 12Z) -9, 12-Octadecadienoate
43 93.605 8.05  43 93.605 8.05
ethyl  ethyl
44 98.297 1-Eicosanol 0.5  44 98.297 1-Eicosanol 0.5
1,2- 1,2-
45 100.562 0.18 45 100.562 0.18
Benzenedicarboxylicacid,dibutylester  Benzenedicarboxylicacid, dibutylester
46 110.209 n-Hexadecanoic acid 7.36 10.4 7.67  46 110.209 n-Hexadecanoic acid 7.36 10.4 7.67
47 128.877 8-Heptadecene 6.43  47 128.877 8-Heptadecene 6.43
48 128.935 Octadec 9 enoic acid . 8.29 48 128.935 Octadec 9 enoic acid . 8.29
시판 누룩과 전통누룩의 향기성분 분석결과， acet i c ac id ethyl ester와 n- nn Analysis of Flavor Components in Commercial Nuruks and Traditional Nuruks, acet ic ac id ethyl ester and n- nn
Zb hexadecanoic acid가 누룩의 향기성분 중 30~40% 이상으로 가장 많은 비중을 차지하였는데， acetic acid ethyl ester는 천연으로는 파인애플 등 과일 속에 존재하는 향기의 한 상분으로， 포도주나 청주 등에도 미량 함유되어 있는 향기성분이다. n-hexadecanoic acid는 waxy하면서 부드럽고 달콤한 향을 내는 향기성분 인식되는 성분이며， Octadec-9-enoic acid은 시큼한 향을 가지는 향기성분으로 송학누룩의 특징적인 향기상분으로 분석되었고， 4-Ethoxy-benzoic acid ethyl ester은 간장의 구수한 향기를 내는 성분으로 중온 발효누룩와 특징적인 향기성분으로 분석되었다. <실험예 9>중은 발효누룩의 저장성 확인  Zb hexadecanoic acid was the most important component of yeast with more than 30 ~ 40% .Acetic acid ethyl ester is a natural component of fragrance that exists in fruits such as pineapple. It is a fragrance ingredient. n-hexadecanoic acid is a waxy, soft and sweet fragrance that is recognized as a fragrance ingredient. Ethyl ester is an ingredient of soy sauce that has been analyzed as medium fermented malt and characteristic flavor component. Experimental Example 9 confirms the shelf life of fermented yeast
상기 <실시예 1>에서 제조된 중은 발효누룩의 저장 기간에 따른 누룩 효소의 잔존능을 분석하였다. , ' 구체적으로， α-amylase, glucoamylase, acidic protease 활성 및 .당화력 (SP)을 측정하였으며， 상기 <실시예 3>과 동일한 방법으로 수행하였으며， 0， 8 및 12주 저장기간 동안 4°C, 15°C 및 30°C 온도에서 누룩 효소의 잔존능을 분석하였다. 그 결과 하기 표 11 및 도 8에 나타난 바와 같이， α-amylase 활성을 비교한 결과 중온 발효누룩의 저장 12주 효소활성은 4°C 저장조건에서 310 units/g로 저장 0주의 효소활성을 100%로 보았을 때 99.9% 유지되었고， 0.1% 효소가 실활 돠었다. 15°C 저장조건에서는 271 units/g로 87.3% 유지되었으며， 실제 12.7% 효소가 실활 되었고， 30°C 저장조건에서는 257.4 units/g로 83% 유지되었고， 실제 17% 효소가 실활 되었다 (표 11 및 도 8). During the preparation in <Example 1> was analyzed the remaining capacity of the yeast enzyme according to the storage period of the fermented yeast. ' Specifically, α-amylase, glucoamylase, acidic protease activity and . The glycosylation potency (SP) was measured, and was carried out in the same manner as in <Example 3>, and the residual ability of the yeast enzyme at 4 ° C., 15 ° C. and 30 ° C. during 0, 8 and 12 weeks storage period was analyzed. It was. As a result, as shown in Table 11 and FIG. 8, the comparison of α-amylase activity results in storage of 12-week enzyme activity of mesophilic fermented yeast stored at 310 units / g at 4 ° C storage conditions. 99.9% of the enzyme and 0.1% of the enzyme was inactivated. At 15 ° C storage conditions, 871% were maintained at 271 units / g, the actual 12.7% enzyme was inactivated, at 30 ° C storage conditions were maintained at 257.4 units / g, 83%, and the actual 17% enzymes were inactivated (Table 11 And FIG. 8).
Glucoamylase 활성을 분석한 결과, 중온 발효누룩의 저장 12주 효소활성은 40C 저장조건에서 5,136 units/g로 95.2% 유지되었지만 실제 4.8% 효소가 실활 되었다. 15°C 저장조건에서는 5,038과 units/g로 93.4% 유지되었으며， 실제 6.6% 효소가 실활 되었고, 30°C 저장조건에서는 4， 776 units/g로 88.6% 유지되었고， 실제 11.4%의 효소가실활 되었다 (표 11 및 도 8) . Analysis of glucoamylase activity showed that the 12-week enzyme activity of mesophilic fermented yeast was 95.2% at 5,136 units / g under 4 0 C storage condition, but 4.8% of enzyme was inactivated. At 15 ° C storage condition, it was maintained 93.4% at 5,038 and units / g, the actual 6.6% enzyme was inactivated, at 88 ° 6 storage condition was maintained at 4, 776 units / g at 88.6%. 11.4% of the enzyme was inactivated (Table 11 and FIG. 8).
Acidi c protease 활성을 분석한 결과， 중온 발효누룩의 저장 12주 효소활성은 40C 저장조건에서 2 , 776.3 uni ts/g로 94.8% 유지되었고 실제 5. 효소가 실활 되었다. 15°C 저장조건에서는 2 , 708.8 uni ts/g로 92.5% 유지되었으며， 실제 7.5% 효소가 실활 되었,고， 30°C 저장조건쎄서는 2 , 629.9 uni ts/g로 89.8% 유지되었고， 실제 10.2%의 효소가 실활 되었다 (표 11 및 도 8) . Acidic c protease activity analysis showed that the 12-week enzyme activity of mesophilic fermented yeast was maintained at 94.8% with 2, 776.3 uni ts / g at 4 0 C storage condition. At 15 ° C storage condition, 92.5% was maintained at 2, 708.8 uni ts / g, the actual 7.5% enzyme was inactivated, and at 8 ° C at 30 ° C storage condition was maintained at 29.8% 9.9 ts / g. 10.2% of the enzyme was inactivated (Table 11 and FIG. 8).
일반적으로 국내에서 많이 통용되는 당화력을 SP( saccharogeni c power ) 단위로 분석한 결과， 중온 발효누룩의 저장 12주 효소활성은 4°C 저장조건에서 1，025.3 SP로 97. 1%유지되었다. 누룩을 제조한 그 시점에서 분석해보면 2.9% 실활 되었다. 15°C 저장조건에서는 959.2 uni ts/g로 90.9% 유지되었으며， 실제 9. 1% 효소가 실활 되었고， 30oC 저장조건에서는 902.9 uni ts/g로 85.5% 유지되었고， 실제 14. ¾의 효소가 실활 되었다 (표 11 및 도 8) . 이들 효소의 잔존능을 분석한 결과， 12주 (90일)의 저장기간 동안 a-amylase , glucoamylase , acidi c protease , saccharogenic power 모두 4°C 저장조건에서 효소 잔존능이 가장 우수한 것으로 40C > 15°C > 30°C 순으로 효소의 잔존능이 우수하였다. 이는 4°C에서 미생물의 증식 및 효소의 분해 작용이 억제되어 12주의 저장기간에도 효소활성이 90% 이상 유지되는 것으로 생각되어진다. In general, the most commonly used glycosylation in Korea was analyzed by the unit of SP (saccharogenic c power). As a result, the 12-week enzymatic activity of mesophilic fermented yeast was 97. 1% at 1,025.3 SP at 4 ° C storage conditions. At the time of manufacture of yeast, analysis was 2.9% inactivated. At 15 ° C storage condition, 90.9% was maintained at 959.2 uni ts / g, and actual 9.1% enzyme was inactivated, and 85.5% was maintained at 902.9 uni ts / g at 30 o C storage condition. Was inactivated (Table 11 and FIG. 8). During storage of the results, and 12 weeks (90 days) was analyzed and the residual capacity of these enzymes, a-amylase, glucoamylase, acidi c protease, saccharogenic power best to 4 0 C> 15 capability both enzyme remaining at 4 ° C Storage Condition The residual ability of the enzyme was excellent in the order of ° C> 30 ° C. It is thought that the proliferation of microorganisms and the degradation of enzymes are inhibited at 4 ° C., so that the enzyme activity is maintained at 90% or more even during the storage period of 12 weeks.
【표 11】  Table 11
온도 (°c)  Temperature (° c)
저장기간 (주)  Storage period
4 15 30  4 15 30
a-amylase 0 310.28± 14.371) 310.28士 14.37 310.28士 14.37 a-amylase 0 310.28 ± 14.37 1) 310.28 士 14.37 310.28 士 14.37
(units/g, 8 . 309.17± 11.08 290.30土 14.90 278.69± 11.03  (units / g, 8.309.17 ± 11.08 290.30 土 14.90 278.69 ± 11.03
weeks)  weeks)
12 310.03±3.11 271.00±9.06 257.44±6.58  12 310.03 ± 3.11 271.00 ± 9.06 257.44 ± 6.58
Glucoamylase 0 5392±90.07 5392±90.07 5392士 90.07  Glucoamylase 0 5392 ± 90.07 5392 ± 90.07 5392 士 90.07
(units/g, 8 5230士 81.02 5258±86.56 5160±32.79  (units / g, 8 5230 士 81.02 5258 ± 86.56 5160 ± 32.79
weeks) 12 5136±76.91 ' 5038±77.78 4776± 10.49 weeks) 12 5136 ± 76.91 '' 5038 ± 77.78 4776 ± 10.49
Acidic 0 2928.52±89.81 2928.52±89.81 2928.52±89.81  Acidic 0 2928.52 ± 89.81 2928.52 ± 89.81 2928.52 ± 89.81
protease 8 2921.48士 8.63 2859.58±71.97 2779.12土.2그 01  protease 8 2921.48 士 8.63 2859.58 ± 71.97 2779.12 °.
(units/g, 12 2776.32±21·70 2708.76±6.85 2629.92±63.79 weeks) (units / g, 12 2776.32 ± 21, 70 2708.76 ± 6.85 2629.92 ± 63.79 weeks)
Saccharogenic 0 1055.64 1055.64 1055.64 power 8 1045.27 1009.25 1002.34 Saccharogenic 0 1055.64 1055.64 1055.64 power 8 1045.27 1009.25 1002.34
(SP, weeks) 12 1025.26 959.15 902.94 (SP, weeks) 12 1025.26 959.15 902.94
값은 평균土 SD(N=3)이다. The value is mean 土 SD (N = 3).

Claims

【청구의 범위】 [Range of request]
【청구항 1】  [Claim 1]
1) 밀을 분쇄하는 단계;  1) milling the wheat;
2) 상기 단계 1)의 분쇄된 밀에 물을 첨가한 후 침윤시키는 단계;  2) adding water to the milled mill of step 1) and then infiltrating it;
3) 상기 단계 2)의 침윤시킨 밀을 성형하는 단계; 3) molding the infiltrated mill of step 2);
4) 상기 단계 3)의 성형된 밀을 34 내지 38°C 온도에서 발효시키는 단계; 및  4) fermenting the shaped mill of step 3) at a temperature of 34 to 38 ° C .; And
5) 상기 단계 4)의 발효 후 건조시키는 단계를 포함하는 중온 발효누룩의 제조방법.  5) Method for producing a medium temperature fermented yeast comprising the step of drying after fermentation of step 4).
【청구항 2] [Claim 2]
제 1항에 있어서， 상기 단계 4)의 발효는 36 내지 37°C 온도에서 발효시키는 것을 특징으로 하는 중온 발효누룩의 제조방법. The method of claim 1, wherein the fermentation of step 4) is a method for producing a medium-temperature fermentation nuruk, characterized in that the fermentation at a temperature of 36 to 37 ° C.
【청구항 3】 [Claim 3]
제 1항에 있어서， 상기 단계 4)와 발효는 1일 내지 30일인 것을 특징으로 하는 중온 발효누룩의 제조방법. The method according to claim 1, wherein the step 4) and the fermentation are 1 to 30 days.
【청구항 4】 [Claim 4]
제 1항에 있어서， 상기 단계 5)의 건조는 40 내지 50oC 온도에서 5 내지 10일 동안 건조시키는 것을 특징으로 하는 중온 발효누룩의 제조방법. The method of claim 1, wherein the drying of step 5) is a method of producing fermented yeast at room temperature, characterized in that for 5 to 10 days to dry at 40 to 50 ° C temperature.
【청구항 5】 [Claim 5]
제 1항의 중온 발효누룩 제조방법으로 제조된 누룩. Nuruk prepared by the method of producing fermented yeast of claim 1.
【청구항 6】 [Claim 6]
제 5항에 있어서， 상기 누룩은 효소활성 및 저장성이 우수한 것을 특징으로 하는 u 중온 발효누룩 제조방법으로 제조된 누룩. The method of claim 5, wherein the yeast is characterized in that the enzyme activity and storage capacity is excellent u Nuruk produced by the method of producing medium-temperature fermented yeast.
【창구항 7】 [Window 7]
제 5항의 누룩으로 제조된 탁주. Takju prepared by the yeast of claim 5.
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