KR101619063B1 - Aspergillus oryzae s-2 kacc93172p of ferment spawn used in brewing rice-wine, and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a flavor and rice wine balhyoje seed to be used in brewing Aspergillus duck No. S-2 (Aspergillus oryzae S- 2) KACC93172P, rice wine balhyoje manufacturing method using the same, and their culture methods, more particularly to a flavor Aspergillus kawachi ( Aspergillus kawachi), which has been widely used as a fermentation agent which has a great influence on the quality of the mungolli brewing in order to produce a quality rice wine having various scents such as fruit flavor, kawachii ) was used instead of aspergillus syringe S-2 ( Aspergillus oryzae S-2) KACC93172P is selected to produce a makgeolli having unique sour taste and unique flavor, and a method for culturing the makgeolli.
Aspergillus oryzae S-2 KACC93172P used as an fermentation agent for fermentation of the makkolli according to the present invention, a Makkolli fermentation product using the same and a method for culturing the fermented Makkolli fermentation product are aspherical fermenting agent Aspergillus kawachi Aspergillus kawachii) than Aspergillus duck low and the composition of organic acids, amino acids are rich harmony of flavors made, with various flavor the S-2 (Aspergillus oryzae S-2) KACC93172P is used as an fermentation agent, it can provide stable basis for stable demand of rice wine, and can provide a basis for scientific and diversification, and further contribute to worldwide expansion.

Description

(ASPERGILLUS ORYZAE S-2 KACC93172P OF FERMENT SPAWN USED IN BREWING RICE-WINE, AND MANUFACTURING USING INGREDIENT USING INGREDIENT USING INGREDIENT BREAKING RICE-WINE, S-2 (Aspergillus oryzae S-2) KACC93172P, METHOD THEREOF}

The invention balhyoje seed used for sake brewing Aspergillus duck No. S-2 (Aspergillus oryzae S- 2) KACC93172P, relates to a rice wine balhyoje manufacturing method using the same, and more particularly, flavor, and various flavor such as fruit flavor ( Aspergillus oryzae S-2 ( Aspergillus oryzae) instead of Aspergillus kawachii, which was conventionally applied as an fermentation agent, oryzae S-2) KACC93172P is selected and applied to the brewing of makgeolli, it is possible to produce rice wine containing harmonious, abundant and unique flavor unlike the makgeolli which is brewed using aspergillus kawachi fermentation agent, And to a process for producing the same.

Makgeolli is the oldest alcohol in history in Korea, and its color is as white as hot, with a low alcohol content of 6 ~ 7 degrees. It has low alcohol content and can be enjoyed freely. Yeast and lactobacillus are alive. Food fiber, vitamins, amino acids and other nutritional aspects are recognized as well-being, and they are getting good response from consumers. In addition, makgeolli is a health food that prevents adult diseases and eliminates thirst and hunger. The richness, refreshing feeling, unique fragrance and rich nutrients produced during the fermentation process have been recognized without distinction of the times.

Recently, as the function of makgeolli began to be recognized, it is becoming popular not only in Korea but also in other countries such as Japan and China, and the export volume is increasing rapidly. However, as makgeolli is expected to slow down in 2012 and go into a stagnant period, the desire for sustained growth through quality upgrading of makgeolli is increasing. In addition, although interest and consumption of makgeolli are increasing, makgeolli still can not be firmly established as a single industry because low-priced products are mainly distributed due to the small size and uniform manufacturing method of the company.

In 2011, the volume of makgeolli increased by 8% to 440,500 보다 (2010 ㎘ 2000 ‰), the only growth in the overall liquor market. In this way, basic and basic research on makgeolli is needed to increase stable demand of makgeolli, not to be ephemeral, but to increase scientific diversification and global expansion.

In traditional makgeolli, traditional yeast is used as the only fermenting agent. Traditional nuruk is mainly made from wheat, and cereals such as corn, barley, soybean, oats, barley and white rice are used depending on the region. The form of the fermented product is a flour made of rice flour, which is made from rice flour, a soup made by scattering grain grains, a powdered flour, a liquid soup made of liquid, ). The most important characteristic of our yeast is the presence of Aspergiluus genus Absidia and Rhizopus genus. The activity of enzymes produced by koji microorganisms and the productivity of organic acids are different depending on the type of koji, raw materials as well as the regions where koji are produced, and there are differences in the quality of flavor, aroma, and color.

At present, fermentation products produced by cultivating Aspergillus kawachii in many breweries are used for brewing makgeolli, and the makgeolli using the fermentation product is too acidic, lacks the amino acid and does not harmonize the taste, none. For this reason, it is evaluated that it causes a problem of deteriorating quality of Takju. Therefore, it is necessary to use differentiated fermentation agents in order to differentiate the different products so that the characteristics of brewers can be exited from the uniform makgeolli production. Therefore, there is a desperate need for studies on the microbiological aspects and enzymatic aspects of the yeast. Therefore, research on the preparation of new fermentation products and the brewing characteristics of the fermentation products are in short supply, and the excellent mold is separated from the yeast, It should be used in the future.

Accordingly, in the present invention, a method of manufacturing a new fermentation agent capable of replacing white germ with traditional koji and selecting a seed culture and culturing it in large quantities is disclosed.

1. Application No. 10-2011-0122160 Manufacturing Method of Improved Nuruk for Makgeolli

1. Park JH, Bae SM, Yook C, Kim JS. Fermentation characteristics of Takju prepared with old rice. Korean J. Food Sci. Technol., 36: 609-615 (2004) 2. Joung EJ, Paek NS, Kim YM. Studies on Korean Takju using the By-Product of Rice Milling, Department of Food & Biotechnology. Food & Biotechnology Research Center, Hankyoung National Univ, Korean J. Food & Nutr., 17: 199-205 (2004) 3. Lee JO, Kim CJ. The effect of adding buckwheat sprouts on the fermentation characteristics of Yakju. Korean J. Food Culture. 26: 72-79 (2011) 4. Han EH, Lee TS, Lee DS. Quality characteristics in mash of Takju prepared by using different Nuruk during fermentation. Korean J. Food Sci. Technol., 29: 555-562 (1997)

The invention as a significant impact on rice wine brewing when jujil balhyoje, the release of this prior application Aspergillus Kawachi (Aspergillus kawachii ) was used instead of aspergillus syringe S-2 ( Aspergillus oryzae S-2) KACC93172P is selected, and it is possible to produce makgeolli which is harmonious, rich and unique flavor different from makgeolli which is brewed using aspergillus kawachi fermentation agent which is applied to most of the distribution makgeolli products. And a method for producing the same.

Above object of the present invention, separated from traditional malt with low organic acid composition it is accomplished by providing a taste and flavor balhyoje Aspergillus duck No. S-2 (Aspergillus oryzae S- 2) KACC93172P used for superior rice wine brewing.

The above-mentioned Aspergillus oryzae S-2 ( Aspergillus oryzae S-2) KACC93172P is achieved by providing Aspergillus oryzae S-2 KACC93172P, which has the nucleotide sequence of SEQ ID NO: 1.

The above object of the present invention can also be achieved by a method for producing rice as described above, comprising the steps of immersing rice for 2 to 3 hours, removing moisture, preparing a boiled rice at 100 ° C for 2 to 3 hours, 50g each aseptically, S-2 ( Aspergillus oryzae S-2) KACC93172P seedlings in an amount of 0.05 to 1% by weight relative to the weight of the increase, and then culturing in a constant temperature and humidity chamber at a temperature of 25 to 30 DEG C and a humidity of 50 to 70% for 24 hours Aspergillus < RTI ID = 0.0 > S-2 < / RTI & oryzae S-2) KACC93172P as an inoculum.

Also above object of the present invention, the Aspergillus duck prepared as described above, the S-2 (Aspergillus oryzae S-2) inoculating the medium with 0.1% to 0.5% by weight of the fermentation agent to be produced as KACC93172P seed culture, adding 3% of bran to the total weight of the medium, adding the bran- Incubating for 48 hours to 70 hours under the culture condition of 50 占 폚 and 80% humidity. Aspergillus oryzae S-2 ( Aspergillus oryzae S-2) KACC93172P as an inoculum.

The aspergillus oryzae S-2 ( Aspergillus sp.), Which is characterized in that the above fermentation agent is dried and pulverized after drying to less than 10% oryzae S-2) KACC93172P as an inoculum.

In addition, the above-mentioned medium may be a bran medium or a PDA medium. Aspergillus oryzae S-2 ( Aspergillus oryzae S-2) KACC93172P as an inoculum.

The above-mentioned Aspergillus oryzae S-2 ( Aspergillus oryzae the S-2) Aspergillus duck No. S-2 (Aspergillus oryzae S-2), characterized in that balhyoje as KACC93172P seed is selected by any one form of the entry form, spore form or in liquid culture form KACC93172P The method comprising the steps of:

Aspergillus oryzae S-2 KACC93172P, an fermenting agent used for brewing a makgeolli according to the present invention, and a makgeolli brewed using a makkolli fermentation agent using the same were used for Aspergillus ( Aspergillus) kawachii ) It has lower composition of organic acid than makkolli which is brewed by using fermentation agent, harmonizes taste with abundant amino acid and contains various flavors. It can provide stable basis for steady increase of demand for makkolli, It also has the effect of contributing to the expansion of the world.

Taste analysis result by taste sensor

Hereinafter, the present invention will be described in more detail.

In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear. In addition, the terms described below are defined in consideration of the functions of the present invention, which may vary depending on the intention of the user, the operator, or the custom.

Therefore, it goes without saying that the definition should be based on the contents throughout this specification.

Example 1. Materials of the present invention

The rice used in the present invention was rice purchased from Nonghyup (Hanaro Mart) in Seokjeong-dong, Anseong-si, Gyeonggi-do. The wheat bran was purchased from Samhwa Mills Co., Ltd. The commercial fermentation agent used as a control was Aspergillus kawachii ) seed product, which was named H-1.

Example 2. Raw material analysis

According to the AOAC (Association of Official Analytical Chemists) method, the moisture was determined by the atmospheric pressure drying method at 105 ° C, the soxhlet extraction method using crude fat, and the semi micro kjeldahl method using crude protein.

Example 3 Preparation of Fermentation Agent

The rice is immersed for 2 hours to 3 hours, the water is removed for 30 minutes to 40 minutes, and the mixture is heated at 100 ° C for 2 hours using a warmer (MODEL-1 Bluebrewlab, Korea). The moisture content is taken to a jeungmi increase to 28 to 32%, based on the weight of the total weight of the sterile by 50g placed in a conical flask 500㎖ Aspergillus duck No. S-2 (Aspergillus oryzae S-2) KACC93172P seedlings were inoculated with 0.05 to 1% by weight of the weight of the increase in weight using platinum, sealed with a foil, shaken, mixed and incubated in a thermostat (SE-96HP, SANG WOO Scientific, Korea) (SE-96HP, SANG WOO Scientific, Korea) with a humidity of 50 to 70% for 24 hours. The Aspergillus oryzae S-2 KACC93172P used as the fermentation agent of the present invention was sensitive to temperature and died under the condition of 30 ° C or higher.

Example 4. Fermentation agent culture

4-1. Seed culture

A PDA (potatodextrose agar) medium was used for the fungal isolation and a Czapek-Dox broth medium was used for the separation of the microorganisms. Aspergillus < RTI ID = 0.0 > oryzae S-2) To investigate the optimum culture medium of KACC93172P and to determine the optimal growth conditions for each medium, PDA medium, Czapek-Dox broth (with 2% Agar), bran medium (bran 10% + agar 2% added) was used. Aspergillus oryzae S-2 KACC93172P was inoculated on each medium and cultured at 27 ° C in an incubator (DR-131, Dreng Science, Korea).

4-2. Fermentation agent culture

Aspergillus < RTI ID = 0.0 > oryzae S-2) KACC93172P was cultured for 24, 48, 72, 96 hours at 25, 30, 35 ℃ and 80% Respectively.

4-3. Inoculation method of fermentation agent

The difference of enzyme activity according to inoculation method was investigated by different forms of fermentation agent. The fermentation agent prepared in the form of spore and the form of entry was dried in a vacuum dryer (OV-01, jeio Tech, Korea) to a moisture content of less than 10% and then pulverized into a pulverizer (Philips, HR-2870, China) Was inoculated in an amount of 0.1, 0.5 wt. In addition, Aspergillus < RTI ID = 0.0 > oryzae S-2) KACC93172P was inoculated in an amount of 1.0% by weight based on the weight of the increase in weight, and the difference in the type of fermentation agent was cultured in a constant temperature and humidity incubator for 3 days.

4-4. Raw material for culture

Aspergillus < RTI ID = 0.0 > oryzae S-2) KACC93172P was inoculated in an amount of 0.1 to 0.5% by weight based on the weight of the increase, and the bran was further added in an amount of 1 to 3% by weight based on the total weight of the fermentation product. The effect of addition of wheat bran as a culture medium was confirmed by incubating for 3 days under constant temperature and humidity condition.

Example 5: Conventional fermentation agents Aspergillus kawachii and Aspergillus kawachii The fermenting agent of the present invention, aspergillus oryzae S-2 ( Aspergillus oryzae S-2) Comparison of characteristics of rice wine sake brewed with KACC93172P

5-1. immersion

Conventional fermentation agents such as Aspergillus kawachii and Aspergillus kawachii The fermenting agent of the present invention, aspergillus oryzae S-2 ( Aspergillus oryzae S-2) KACC93172P, respectively, in order to compare the characteristics of each rice wine. The fermentation agent was Aspergillus S-2 oryzae S-2) S-2 cultured with KACC93172P and H-1 cultured with Aspergillus kawachii , which is a common fermenting agent, were used. The main breeder was dipped in La Parisienne (SI, Lesaffre, France) among commercially available yeast. The incubation time was determined by incubating for 48 hours at 23 ° C with shaking for 48 hours. Stage 1 was incubated for 24 hours at 23 ° C, and incubated for 120 hours at 23 ° C. Table 1 shows the ratio of raw materials. After fermentation, fermentation time, temperature, ethanol content, reducing sugar, acidity, pH, yeast count and bacterial count were compared for 8 days.

Seed bacteria Configuration Mother One-step soak Two-stage soaking
S-2
H-1 rice 237 1000 Yeast (g) 12 Water (ml) 225 880 1700 Rice entry (g) 150 237

5-2. Composition of fermentation product

5-2-1. Reduction sugar measurement

The sample was obtained by filtration of the syrup by measuring and quantitating by direct reducing sugar method described in the mainstream analysis regulations. Then, 20 ml of distilled water was added to a 200 ml Erlenmeyer flask, and 5 ml of Fehling A solution and 5 ml of Pelling B solution / RTI > The solution was added just before the work. The Erlenmeyer flask was placed on a hot plate and the Felling mixture was boiled. Then, using a 2-ml pipette, shake the sample until the color changes in the boiled solution, and record the amount of sample used until the color changes. 8 ml of standard glucose solution was added rapidly. When the solution containing the sample starts to boil, add 4 drops of methylene blue solution and shake to observe the color change. Add more standard glucose until the color of the solution changes to purple and use the standard glucose solution Lt; / RTI > If it does not spread quickly, add a standard glucose solution.

The control group value is measured in the same manner as above, and the amount of standard glucose solution used is recorded. Only 10 ml of distilled water was added to the control instead of the sample.

5-2-2. Titratable acidity and pH

The titratable acidity was measured by dropping 2 to 3 drops of the mixed indicator in a 10 ml sample according to the regulations of the National Bureau of Statistics (USA), and neutralizing titration with 0.1 N sodium hydroxide (NaOH). The titration value was converted into lactic acid (%) Respectively. The pH was also measured by a pH meter (model 745P, ISTEK, Korea) according to the IRS mainstream analysis regulations.

5-2-3. Ethanol content

During the fermentation period, the amount of ethanol was changed to 100 ml of distilled water by distillation, 70 to 80 ml, 100 ml of distilled water, and 100 ml of distilled water according to the regulations of the National Liquor Administration. Lt; / RTI >

5-3. Organic acid composition investigation

Organic acids were analyzed by post-column method with HPLC-20A series (SHIMADZU co., Tokyo, Japan). The sample of the makgeolli soup was filtered using a 0.45 μm membrane filter and connected to two SHODEX RSpak KC-811 (8.0 mm ID × 300 mm) columns. The oven temperature was 63 ° C. and the flow rate was 0.8 Ml / min, the mobile phase was 3 mM perchloric acid, and the injection volume was 10 μl. The separated organic acids were developed with 0.2 mM BTB, 15 mM Na2HPO4, and 2 mM NaOH in a reaction coil at a temperature of 25 ° C and a flow rate of 1.0 mL / min. The absorbance at 440 nm was measured Respectively. All experiments were repeated three times.

5-4. Flavor composition of rice wine with different fermentation agents

The aroma composition of rice wine with different fermenting agents was investigated. Volatile components were analyzed using GC-MS (Agilent 6890N GC / Agilent 5973 mass selective detector (MSD, Agilent Co., Palo Alto, CA, USA). The volatile components were extracted with SPME fiber (50/30 μm divinylbenzene, carboxen on polydimethylsiloxane) (Supelco Co., Bellefonte, Pa., USA) and 10 ml of makgeolli sample was placed in a 20 ml headspace vial and sealed with Teflon cap. After reaching an equilibrium state at 50 ° C for 30 minutes, the volatile components were adsorbed for 1 minute by exposing 1 cm of SPME fiber, and the fiber was exposed to the GC injector port (200 ° C) and desorbed for 1 minute. DB-wax (60 m length × 0.25 mm id × 0.25 μm film thickness: J & W Scientific, Folsom, CA, USA) was used for the GC / MS column for the chromatography, and the oven temperature was 40 ° C. For 5 minutes and then heated up to 200 ° C at a rate of 5 ° C / min and maintained for 20 minutes. The injector temperature was 200 ℃, the detector temperature was 250 ℃, helium was used as the carrier gas, and the flow rate was 1.1 ㎖ / min. The ionization voltage was 70 eV and the molecular weight range (m / z) analyzed was 33-500.

6. Sensory Quality Inspection

6-1. Sensory test

The sensory evaluation was carried out on 20 students of food science and biotechnology department of Hankyong University. The sensory evaluation was carried out by using the makgeolli made with commercial fermentation agent and Aspergillus oryzae S-2) Evaluation of color, flavor, taste, and overall acceptability of makgeolli made with fermentation agent using KACC93172P to be 6% ethanol, 0.18% acidity and 15mg / ㎖ aspartame of 100ppm Method.

6-2. Taste Analysis by Taste Sensor

To confirm the difference of taste objectively, we used a taste analyzer (Taste Sensor, Isent, Japan) using a taste sensor. The rice wine was diluted 10 times, and 35 ml of each sample was added to the taste test machine. The taste was measured four times and the average value of the remaining values except the one measured value was used. We analyzed the sourness, bitterness, astringency, umanmi, saltiness and richness as the measurement items. The result of the measurement by the electrode of the sensor is converted into the value of the taste again and is shown as a result. The numerical difference in taste indicates a change in concentration of the taste component that can recognize a difference in taste of a person.

Example 7. Results

7-1. Fermentation agent culture

7-1-1. Seed culture

Effects using a medium prepared directly with a common fungal culture medium on the market on growth, Aspergillus duck the S-2 (Aspergillus oryzae S-2) KACC93172P was incubated at a temperature of 30 ° C and a humidity of 80% for 72 hours to select an optimum medium for the difference in activity of the fermentation broth.

Aspergillus < RTI ID = 0.0 > oryzae S-2) KACC93172P was inoculated on a PDA, wheat bran, and Czapek culture medium, and the activity was measured. Aspergillus oryzae S-2 ( Aspergillus oryzae S-2) in KACC93172P, followed by PDA medium 98.93 sp, bran medium 77.59 sp, and Czapek medium 86.23 sp. Protein degradation analysis showed 39.18 and 39.98ml casein / h in PDA and wheat media, respectively, but lower in casein / h in Czapek medium than 18.87ml casein / h. The acidity was 0.10-0.30 ml and the moisture content was 28.33-32.56%. These results indicate that the strain preserving medium exhibits higher saccharifying activity than the Czapek mold base medium in PDA medium and wheat bran medium. In addition, it was confirmed that as the nutrient condition was abundant in the medium, the activity of the strain was transferred to the culture medium.

Aspergillus oryzae S-2 KACC93172P ( Aspergillus oryzae S-2) S-2




30 ℃







PDA
SP 98.93 + - 8.53 ^a PA 39.18 ± 0.60 ^a acid 0.30 0.10 ^a MC 28.33 ± 1.20 ^a

CZA
SP 77.59 ± 0.87 ^a PA 18.87 + 0.44 ^a Acid 0.10 0.03 ^c MC 32.38 ± 2.81 ^a
Bran
juice SP 86.23 ± 0.88 ^a PA 39.98 ± 1.01 ^a Acid 0.10 0.05 ^a MC 32.56 ± 2.28 ^a

7-1-2. Incubation time

Aspergillus < RTI ID = 0.0 > oryzae S-2) KACC93172P in order to find the optimal culture conditions, the enzyme activity according to the incubation time with different incubation temperature at 80% humidity is shown in Table 3, Table 4 and Table 5.

According to the experimental results, the activity of glycosylation enzyme increased sharply after 48 hours, but protease activity and acidity were not measured until 48 hours after the initiation of culture.

24 48 72 96 Protease activity 25 ℃ S-2 - - 39.21 + - 0.14 a ^b 78.75 + - 0.84 ^a 30 ℃ S-2 - - 39.18 ± 0.40 ^ab 79.79 ± 0.61 ^a 35 ℃ S-2 - - 39.96 ± 0.48 ^ab 79.79 + 0.64 ^a

24 48 72 96 Acidity 25 ℃ S-2 - - 0.20 ± 0.08 ^b 0.30 ± 0.05 ^abc 30 ℃ S-2 - - 0.15 0.03 ^ab 0.20 ± 0.06 ^c 35 ℃ S-2 - - 0.15 0.03 ^ab 0.20 ± 0.08 ^bc

24 48 72 96 Moisture content 25 ℃ S-2 32.05 + 0.63 ^a 32.16 +/- 1.47 ^a 32.04 + 0.66 ^a 30.34 + 1.03 ^a 30 ℃ S-2 31.95 +/- 1.43 ^a 32.34 + 1.82 ^a 32.26 ± 1.92 ^a 32.45 + 1.16 ^a 35 ℃ S-2 32.79 ± 2.94 ^a 33.37 ± 1.55 ^a 33.58 ± 2.49 ^a 33.31 + 1.63 ^a

7-1-3. Inoculation method of fermentation agent

The results of the inoculation type of fermentation agent showed that the fermentation agent was more easily absorbed in the incubation group than that in the inoculated experimental group. Also, it was effective to inoculate the inoculation amount of the fermentation agent in an amount of 0.1 to 0.5% by weight based on the weight of the increased amount when directly inoculating the fermentation agent based on preliminary experiments. As a result of the inoculation of the fermenting agent by the mode of entry, it was confirmed that it can be used as a seed when the S-2 ( Aspergillus oryzae S-2) KACC93172P is manufactured as an entry form (Table 6).

The results of inoculation in the form of spores are shown in Table 7. The spore morphology also had no problems in the form of the fermentation agent and in the preparation of the fermentation agent. However, the inoculation method of spore type was more active than the inoculation type. The liquid form inoculum was aspergillus syringe S-2 ( Aspergillus oryzae S-2) KACC93172P There was no significant difference in the activity of saccharifying enzyme according to the amount of liquid inoculum. Therefore, in the present invention, 1% by weight liquid seeds were inoculated to the weight of the weight.

Results for liquid inoculation (Table 8) confirm that there is no problem in producing fermentation products in different forms of seeds. As for the specificity of the strains based on the finished products, the method of inoculating the aseptically inoculated strains was safe for the manipulation of the strains and the contamination. In the convenience and final storage period, the fermentation agent and spore culture method were the best methods .

A. oryzae S-2 0.1 0.5







Entry Form
Fermentation agent







25 ℃
SP 62.12 + 4.13 ^a 63.26 ± 1.56 ^a PA 19.53 ± 0.2 ^a 3 19.03 ± 0.67 ^a acid 0.10 0.03 ^a 0.10 0.05 ^a MC 30.81 + 1.97 32.25 + 1.33 ^b

30 ℃
SP 85.70 ± 3.28 ^a 79.03 ± 6.15 ^a PA 39.61 + 0.18 ^a 38.56 ± 1.27 ^a acid 0.15 + 0.05 ^abc 0.05 ± 0.06 MC 30.57 ± 2.25 ^a 31.44 + 1.04 ^a

35 ℃
SP 72.41 + 4.76 ^ab 80.83 + - 3.51 ^a PA 19.70 ± 0.61 ^b 39.01 + 0.36 ^a acid 0.20 ± 0.05 ^a 0.10 0.03 ^a MC 30.87 ± 1.17 ^a 31.91 + 0.68 ^a

A. oryzae S-2 0.1 0.5








Spore morphology
Fermentation agent







25 ℃
SP 92.94 ± 5.38 ^a 85.36 ± 4.17 ^a PA 39.18 ± 0.57 ^a 38.59 + 0.37 ^a acid 0.12 + 0.03 ^a 0.12 + 0.03 ^a MC 32.25 + 1.05 ^a 30.98 + 0.72 ^b

30 ℃
SP 154.77 ± 13.71 ^b 157.02 + - 6.54 ^a PA 79.92 + 0.83 ^a 78.94 ± 0.53 ^a acid 0.25 0.03 ^ab 0.31 0.08 ^a MC 33.52 ± 1.05 ^a 30.68 ± 2.60 ^a

35 ℃
SP 131.99 ± 6.46 ^ab 138.781 ± 0.99 ^a PA 76.36 ± 0.60 ^b 78.60 ± 0.37 ^a acid 0.24 + 0.01 ^a 0.25 0.03 ^ab MC 31.99 + 2.06 ^a 38.82 ± 5.15 ^a

A. oryzae S-2 1.0





Liquid culture form
Fermentation agent







25 ℃

SP 51.36 ± 2.10 ^b PA 18.96 + 0.17 ^b acid 0.30 0.01 ^b MC 32.72 ± 1.21 ^a

30 ℃
SP 105.94 + 4.15 ^a PA 79.90 ± 0.21 ^a acid 0.32 ± 0.03 ^a MC 33.80 + 1.51 ^a
35 ℃

SP 65.42 ± 1.18 ^b PA 19.26 ± 0.16 ^b acid 0.32 ± 0.03 ^b MC 31.46 ± 1.03 ^a

7-1-4. Raw material for culture

The bamboo shoots and wheat bran were mixed to prepare Aspergillus strain S-2 oryzae S-2) KACC93172P was inoculated and cultured at 25, 30, and 3 ° C for 72 hours based on 80% humidity, and the activity of each of the fermentation agents was measured (Table 9). As a result of the experiment, it was observed that when the fermentation broth was added to the bran at the boiling point, the spore formation time of the fermentation agent appeared very rapidly and was formed vigorously. Thus, it was found that the acidity and especially the proteolytic ability I could confirm. As a result, the higher the amount of bran added, the higher the sugar content was.

A. oryzae S-2 Bran concentration 0.1 3.0
25 ℃

SP 47.50 + 4.98 ^b 56.62 ± 4.86 ^b PA 39.31 + - 0.52 ^a 38.95 + - 0.53 ^a acid 0.30 0.03 ^abc 0.25 ± 0.03 ^bc MC 31.96 ± 3.17 ^a 33.5 ± 1.80 ^a

30 ℃
SP 193.80 ± 10.93 ^b 272.46 +/- 12.35 ^a PA 79.35 ± 1.08 ^a 78.79 ± 0.37 ^a acid 0.25 0.08 ^a 0.15 + 0.03 ^b MC 31.82 ± 1.14 ^ab 32.88 ± 2.35 ^ab

35 ℃
SP 246.49 + - 13.03 ^b 323.91 ± 8.10 ^a PA 68.23 ± 1.32 ^bc 75.61 ± 2.45 ^a acid 0.10 0.03 ^c 0.25 ± 0.05 ^bc MC 32.89 ± 2.83 ^a 32.16 ± 1.00 ^a

7-2. Comparison of Fermentation Characteristics of Makkoli Brewed with Different Fermentation Agents

Aspergillus < RTI ID = 0.0 > oryzae S-2) The results of the total of 8 days after immersion of the experimental group S-2 made with KACC93172P in one step (Table 10) and commercial products such as Aspergillus kawachii ). The results of the total 8 days of incubation of H-1 (Table 11) were confirmed.

All of the two cultures were incubated with caution not to exceed 25 ℃.

The pH change was maintained in the range of 4.0 to 5.0 in the fermentation medium S-2 of the present invention and the fermentation was maintained in the range of 3.0 to 5.0 in the commercial fermentation medium H-1 to maintain the H-1 fermentation agent lower than that of the S-2 fermentation agent .

The acidity of the S-2 fermentation product of the present invention was found to be in the range of 1.0 to 2.8 and in the case of the commercial fermentation product H-1, it was found to be 4.9 to 9.2. Respectively. According to the conventional report of Park et al. (1), it is reported that the change of the acid content in the snack of Takju is small, and the abnormal fermentation is considered to occur when the acid content is increased sharply. According to this report, Aspergillus oryzae S-2 was confirmed to be maintained at 1.0 to 3.0 without significant change in the acid content than the commercial fermentation product H-1, It was confirmed that KACC93172P has an excellent acidity as a rice wine fermentation agent. In addition, a suitable acid has a positive aspect to inhibit the propagation of the germs to improve the taste and to ferment the germination. In addition, according to Jung et al. (2), acidity has been reported to increase due to the action of microorganisms that grow on the fungus, and similar results as the experimental results of the present invention were ascertained.

In the reducing sugar content, the fermentation time of the fermentation medium S-2 and the fermentation medium H-1 of the present invention was high at 17.03 mg / ml and 10.18 mg / ml on the first day, respectively. However, The values of S-2 and M-1 were 1.01 mg / ml and 1.95 mg / ml, respectively. (3) and (4) that the reducing sugar is lowered in the latter stage of fermentation because it is converted to ethanol by the yeast as the fermentation progresses. In addition, in the report of Park et al. (1), the reduction of reducing sugar during fermentation of syrup is closely related to the number of yeast and the production of ethanol. Especially, during the fermentation process, the reduction of reducing sugar And it was confirmed that the concentration of ethanol was increased proportionally.

The content of ethanol was found to be similar to that of the fermentation product S-2 of the present invention and the commercial fermentation product H-1. The ethanol content of the first day showed a low value of 6.3% in S-2 and 6.6% in H-1, and gradually increased with the first day, 17.4% of S-2 and 18.1% of H- %. It was confirmed that the concentration of ethanol was increased in proportion to the reduced amount of reducing sugar identified above.

analysis Product temperature (℃) pH The acidity (ml) Reducing sugar (mg / ml) Ethanol content (%) 1 day 22.0 4.851 1.1 17.03 6.3 2 days 25.2 4.431 1.0 26.91 8.2 3 days 22.5 4.356 1.7 6.64 11.7 4 days 23.1 4.399 2.0 7.28 14.0 5 days 20.6 4.417 2.2 1.73 15.7 6 days 23.9 4.437 2.3 1.34 16.0 7 days 22.4 4.490 2.3 2.12 16.4 8 days 22.4 4.510 2.8 2.01 17.0 9th 22.6 4.600 2.6 1.01 17.4

analysis Product temperature (℃) pH The acidity (ml) Reducing sugar (mg / ml) Ethanol content (%) 1 day 22.5 4.32 9.2 10.18 6.6 2 days 24.5 3.59 4.9 24.17 8.1 3 days 22.3 3.62 5.7 4.14 11.7 4 days 23.3 3.68 5.9 2.55 14.0 5 days 20.4 3.75 6.6 1.58 15.3 6 days 23.9 3.81 6.8 1.33 16.3 7 days 22.1 3.95 6.6 4.56 16.5 8 days 22.4 4.01 7.1 4.78 17.6 9th 22.7 4.07 6.7 1.95 18.1

In the overall aspect, the initial reducing sugar content was higher in the fermenting agent S-2 of the present invention than in the H-1, and the fermentation agent of S-2 showed higher acidity and pH than the H-1 fermentation agent in the latter stage of fermentation, The content and the reducing sugar content did not show any significant difference.

7-3. Organic acid composition

The organic acid of the makgeolli sample was analyzed by HPLC and is shown in Table 12.

division Oxalic acid
(Oxalic
acid) Citric acid
(Citric
acid) Malian
(Malic
acid) Succinic acid
(Succinic
acid) Lactic acid
(Lactic
acid) Acetic acid
(Acetic
acid) Aspergillus kawachii ) 3.3 3492.3 460.4 914.7 493.8 87.9 Aspergillus oryzae S-2 KACC93172P ( Aspergillus oryzae S-2) 0.5 664.3 168.7 738.8 367.1 3.3

As a result of the analysis, it was found that oxalic acid, malic acid, succinic acid, acetic acid, Aspergillus kawachii ), it was possible to confirm low organic acid composition. Through this, the mountainous Aspergillus kawachii ) with Aspergillus < RTI ID = 0.0 > oryzae S-2) KACC93172P as a fermentation agent.

7-4. Flavor composition of rice wine with different fermentation agents

Table 13 shows the composition of the aroma components of the rice wine of each fermented product identified in Example 5-4.

compound Aspergillus kawachii ) Aspergillus oryzae S-2 KACC93172P ( Aspergillus oryzae S-2) Average SD Average SD Ethyl acetate 3.85 0.03 3.36 0.25 2-methylpropyl acetate 0.36 0.01 0.73 0.12 Ethyl butanoate 0.28 0.05 0.29 0.04 1-Propanol 0.52 0.19 0.17 0.05 2-ethyl-1-propanol 1.09 0.14 1.55 0.12 3-methyl-1-butanol acetate 2.95 1.47 6.56 2.44 Isoamyalcohol (3-methyl-1-butanol) 7.59 1.02 7.64 - Ethyl Hexanoate 0.58 0.06 0.90 0.05 Ethyl Octanoate 4.13 0.89 6.87 0.37 3-methylbutyl octanoate - - 0.18 0.03 Ethyl decanoate 6.46 1.53 10.62 1.47 Ethyl dodecanoate 1.73 1.32 3.21 0.68 2-Methoxy-4-vinylphenol - - 0.94 0.14 2-phenylethyl acetate 3.06 2.45 2.26 0.20 Ethyl tetradecanoate 1.31 0.97 1.75 0.39 Ethyl oleate 0.58 0.21 0.62 0.04 Ethyl linoleonate 0.95 0.05 0.94 0.10

As a result of the analysis, it was found that Aspergillus kawachii , Aspergillus oryzae S-2 ( Aspergillus oryzae S-2) 3-methyl-1-butanol acetate corresponding to banana or orientation in KACC93172P, Ethyl hexanoate corresponding to pineapple or banana or wine, Ethyl octanoate corresponding to orange pineapple or berries, Peach, Pineapple, Ethyl dodecanoate, which is equivalent to Ethyl decanoate, Fruit flavor or Flower color,

7-5 Sensory Quality

7-5-1. Sensory test

Aspergillus < RTI ID = 0.0 > oryzae S-2) The fermentation agent ( Aspergillus kawachii ), which is inoculated with the fermentation product of the present invention inoculated with KACC93172P , is shown in Table 12.

The color, flavor, taste, and overall quality shown in Table 14 are the items that can confirm the preference of the product, and fruits, feces, and alcohol are items that can confirm the strength of the product.

Aspergillus oryzae S-2 of the present invention ( Aspergillus oryzae) than the products using the fermentation products commonly used in fragrance, taste, oryzae S-2) were able to see the high scores in the sensory evaluation of products used to KACC93172P balhyoje, Aspergillus duck in the other Fruit items invention takes the S-2 (Aspergillus oryzae S-2) KACC93172P. As a result, it was found that aspergillus oryzae S-2 ( Aspergillus oryzae S-2) KACC93172P was inoculated with a large amount of metabolites.

Sensory properties A. kawachii Entrance A. oryzae KACC93172P
Entrance color 6.80 ± 1.54 5.50 ± 1.91 incense 4.50 ± 1.40 6.80 ± 1.36 flavor 4.65 ± 1.73 5.35 ± 2.13 Overall quality 4.10 ± 1.45 6.45 ± 1.61 Fruitlessness 4.35 ± 0.99 5.60 ± 1.82 Inside 5.50 ± 1.91 4.95 ± 1.50 Alcoholism 6.00 ± 1.91 4.80 ± 1.44

7-5-2. Taste Analysis by Taste Sensor

In order to confirm the difference in objective taste, an fermentation product made of Aspergillus oryzae S-2 KACC93172P isolated from a conventional yeast using a taster sensing system and a fermentation product made of a commercial fermentation product The taste of makgeolli made with H-1 was analyzed.

Analysis of five kinds of makgeolli was shown in Table 15 and Fig. As a result of the overall taste analysis, the commercial fermentation agent Aspergillus In the product group H-1 produced as kawachii , the sour taste and the salty taste were excellent, and the bitter taste, the sweet taste and the tender taste were aspergillus syringe S-2 ( Aspergillus oryzae S-2) KACC93172P as the fermentation agent. As a result, it was confirmed that various flavors could be harmonized by using Aspergillus oryzae S-2 KACC93172P in the makgeolli brewing which desires various flavors.

Sensory properties Makgeolli H-1 SNU-2 Sourness -4.70 ± 0.16 ^{b 2)} -20.44 + 0.28 ^e Bitterness 4.12 ± 0.15 ^e 8.27 ± 0.23 ^a Astringency 1.78 ± 0.01 ^e 2.26 + 0.03 ^a Umami 3.48 ± 0.04 ^e 8.51 + 0.13 ^a Richness 1.07 ± 0.16 ^a 1.29 + 0.30 ^a Saltiness -12.20 + 0.04 ^a -16.11 + -0.13 ^c

Agricultural Biotechnology Research Institute KACC93172 20130219

<110> Gyeonggi Makgeoli Golbalization Association <120> ASPERGILLUS ORYZAE S-2 KACC93172P OF FERMENT SPAWN USED IN          BREWING RICE-WINE, AND MANUFACTURING METHOD THEREOF <130> P14 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 713 <212> DNA <213> Aspergillus oryzae S-2 KACC93172P <400> 1 tgcggaagga tcattaccga gtgtagggtt cctagcgagc ccaacctccc acnccgtgtt 60 tactgtacct tagttgcttc ggcgggcccg ccattcatgg ccgccggggg ctctcagccc 120 cgggcccgcg cccgccggag acaccacgaa ctctgtctga tctagtgaag tctgagttga 180 ttgtatcgca atcagttaaa actttcaaca atggatctct tggttccggc atcgatgaag 240 aacgcagcga aatgcgataa ctagtgtgaa ttgcagaatt ccgtgaatca tcgagtcttt 300 gaacgcacat tgcgccccct ggtattccgg ggggcatgcc tgtccgagcg tcattgctgc 360 ccatcaagca cggcttgtgt gttgggtcgt cgtcccctct ccggggggga cgggccccaa 420 aggcagcggc ggcaccgcgt ccgatcctcg agcgtatggg gctttgtcac ccgctctgta 480 ggcccggccg gcgcttgccg aacgcaaatc atctttttcc aggttgacct cggatcaggt 540 agggataccc gctgaactta agcatatcaa taagccggag gaaagatcat taccgagtgt 600 agggttccta gcgagcccac ctccaccctt gtttaccgta cctcagttgc ttcggcgggg 660 cccccttcat ggccgccggg ggctctcagc cccgggcccg gcccaccaaa aac 713

Claims (7)

(Aspergillus oryzae S-2) KACC93172P having the nucleotide sequence of SEQ ID NO: 1, which is an effective yeast strain used for brewing makgeolli which has a low organic acid composition and is excellent in taste and aroma. delete Immersing the rice for 2 to 3 hours, removing moisture, and raising the rice at 100 占 폚 for 2 to 3 hours;
The above-mentioned steeping was aseptically taken out in an amount of 50 g for the first fermentation to inoculate 0.05 to 1% by weight of Aspergillus oryzae S-2 strain KACC93172P against the weight of the increase, , And culturing in a thermostat with a humidity of 50 to 70% for 24 hours; (Aspergillus oryzae S-2) KACC93172P having the nucleotide sequence of SEQ ID NO: 1, which comprises the nucleotide sequence of SEQ ID NO: 1. A fermentation broth comprising Aspergillus oryzae S-2 KACC93172P having the nucleotide sequence of SEQ ID NO: 1 as prepared in claim 3 is cultivated by secondary fermentation, 0.1 to 0.5% by weight in the culture medium;
Adding 3% bran to the total weight of the culture medium;
Culturing the bran-added culture medium for 48 to 70 hours at a temperature of 30 DEG C and a humidity of 80%; (Aspergillus oryzae S-2) KACC93172P having the nucleotide sequence of SEQ ID NO: 1, which comprises the nucleotide sequence of SEQ ID NO: 1. 5. The method of claim 4,
The aspergillus oryzae S-2 KACC93172P having the nucleotide sequence of SEQ ID NO: 1 is cultivated in the fermentation broth as seed culture, characterized in that the above-mentioned fermentation agent is dried at less than 10% and then pulverized and inoculated. Way. 6. The method of claim 5,
(Aspergillus oryzae S-2) KACC93172P having the nucleotide sequence of SEQ ID NO: 1, wherein the culture medium is a bran culture medium or a PDA (potato dextrose agar dedium) culture medium. A method for culturing an inoculum of an inoculum. 6. The method of claim 5,
The fermentation broth containing Aspergillus oryzae S-2 KACC93172P having the nucleotide sequence of SEQ ID NO: 1 as a seed may be selected from any one of an entry form, a spore form and a liquid culture form (Aspergillus oryzae S-2) KACC93172P having the nucleotide sequence of SEQ. ID.

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