KR101285228B1 - Method of preparing nuruk and fermented alcohol using the same by inoculating Aspergillus oryzae into unpolished-glutinous rice - Google Patents

Abstract

The present invention relates to a yeast and a method for preparing fermented wine using the same. More specifically, a method of preparing yeast by inoculating Aspergillus aurise (Aspergillus oryzae, microorganism accession number: KCTC11922BP) cultured as a strain on brown rice glutinous rice. will be. Nuruk according to the present invention and the fermented wine prepared using the same can provide a fermented wine excellent in appearance, aroma and taste.

Description

     Method of preparing nuruk and fermented wine using the yeast by inoculating Aspergillus duckase as a strain on brown rice glutinous rice {Method of preparing nuruk and fermented alcohol using the same by inoculating Aspergillus oryzae into unpolished-glutinous rice}

The present invention is inoculated with Aspergillus aurise (Aspergillus oryzae, microorganism accession number: KCTC11922BP) cultured as a strain on brown rice glutinous rice, and relates to a method of producing yeast and fermented wine using the yeast.

     Liquor contains the climate and folklore of the country, and has been closely associated with mankind for many years as a representative beverage that reflects history. Our folk liquor uses yeast using various grains (barley, wheat, green beans, corn, etc.), and various grains (barley, corn, sorghum, green beans, crude, buckwheat, red beans, etc.) and medicinal herbs. It was in its heyday as it left the name of more than 360 kinds of alcoholic beverages, but it was mostly cut off during the Japanese colonial period.

However, in the case of related alcoholic beverages, including the recently sold folk liquor, molds and yeasts of Aspergillus genus, which are used to make sake, are mainly used, and raw materials are limited to rice, so the variety of traditionalism is lost and consumers also The folklore market is in recession due to the food crunch.

     In addition, the development of efficient manufacturing technology of traditional medicine and Takju, analysis of various chemical components such as raw materials and rice flour, development of fermenter spawn and distillation characteristics of traditional cultured liquor, distribution and storage of microorganisms and enzymes in yeast and rice flour Extension and quality improvement have been reported. In addition, medicinal liquors in the form of leaching liquor (liqueur wine) using several medicinal plants have been developed. However, research on improving the physicochemical characteristics and physiological (physiological) functional substances and improving the quality of most folk liquors has not been conducted so far, so it is falling behind in quality competitiveness with foreign liquors.

     In addition, our traditional folk liquor is diverse in the distribution of microorganisms that act on the saccharification and fermentation of grains, which leads to various forms of metabolism during the brewing process, making it difficult to produce a uniform quality product.

Accordingly, the present inventors have come to the present invention to separate the excellent spawn suitable for various raw grains to develop yeast and to manufacture folk liquor using various grains.

It is an object of the present invention to inoculate Aspergillus aurise (Aspergillus oryzae, microorganism accession number: KCTC11922BP) cultured as a strain on brown rice glutinous rice to provide a fermented liquor prepared by using the yeast.

     In order to achieve the above object, the present invention provides a method for preparing yeast by inoculating Aspergillus aurise (Aspergillus oryzae, microorganism accession number: KCTC11922BP) cultured as a strain on mung beans.

     In another embodiment, the present invention provides yeast prepared by inoculating brown rice glutinous rice inoculated with a strain of Aspergillus aurise (Microbial Accession No .: KCTC11922BP).

     In yet another embodiment, after inoculating Aspergillus aurise (Aspergillus oryzae, microorganism accession number: KCTC11922BP) cultured as a strain on brown rice glutinous rice to prepare yeast, a method for producing fermented liquor using the yeast To provide.

     In another embodiment, the fermented liquor fermented with yeast prepared by inoculating Aspergillus arige (Aspergillus oryzae, microorganism accession number: KCTC11922BP) cultured as a strain in brown rice glutinous rice.

     In the present invention, the strain may be characterized in that it is cultured in a solid medium consisting of potato medium (PDA, Potato Dextrose Agar) and bran, the strain may also be characterized in that cultured in a liquid medium of bran. .

In the present invention, the strain may be characterized in that cultured in a medium having a water content of 20 to 30.

Fermented wine prepared using yeast prepared by inoculating Aspergillus aurise (Aspergillus oryzae, microorganism accession number: KCTC11922BP) cultured as a strain on brown rice glutinous rice according to the present invention will provide a folk wine excellent in appearance, aroma and taste. Can be.

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

Selection of Strains According to Morphological Characteristics and Types of Grains

1-1. Isolation of mold

In order to separate the mold from the test grains produced and the collected grains and yeasts, grains of yeast and yeast were prepared in 1 g of peptone dilution fluid (0.1% peptone, 0.85% NaCl, 0.03% KH ₂ PO ₄ , 0.04% Na ₂ HPO _4). ) 9ml was added, vortexed for 10 minutes, and allowed to stand. A supernatant was used as a sample. The collected grains were stomached for 1 minute with 45 ml of peptone dilution fluid (0.1% peptone, 0.85% NaCl, 0.03% KH ₂ PO ₄ , 0.04% Na ₂ HPO ₄ ) added for 5 minutes. Dilute each sample with 10 ¹ , 10 ² , 10 ³ , 10 ⁴ , 10 ⁵ , 10 ⁶ , YSC (yeast nitrogen base without amino acids 0.1%, soluble starch 0.5%, congo red 0.05%) and SF (soluble starch fungi selection medium) was plated and cultured for 30 to 3 days. The cultured Nuruk microorganisms were separated through pure passage through 3 to 5 passages and separated, and stored in glycerol stock. Hereinafter, the selection process for the separated microorganism will be described.

end. Separation from Grain Yeast

Grain malt is 19 kinds collected by each region such as ginseng, millet, mung bean, soybean, barley, buckwheat, rice, wheat, sorghum, rice barley, waxy barley, brown rice, glutinous rice, glutinous rice black rice, brown rice, red beans, black rice, yulmu, tea It was made using 144 grains. A total of 405 strains of fungi were isolated from pure yeast by the test production. These strains were classified by morphological characteristics and types of grains and 160 strains were selected.

I. Strain Separation from Grains

Twenty-six fungal strains were screened from isolates of 177 grains, and growth patterns were similar in culture media such as yellow green spores, yellow spores, black spores, white mycelium, and gray mycelia.

All. Separation from Yeast

The fungus was isolated from 289 yeast and 437 strains were isolated. Similar morphology of yellow spores, black spores, white hyphae, yellow green spores, and yellow spores was observed. Among them, 144 strains, depending on their characteristics, such as different mycelium color and shape, and different growth rate Was reselected.

Screening according to strain activity

end. Acid production capacity measurement

     Strains selected from grain yeast, grains and yeast were first plated on soluble starch fungi selection medium (SF) medium and the size of the red rings produced after incubation at 30 was measured to determine acid production capacity.

I. Starch Degradation Activity Measurement

     After inoculation into the starch medium, the pure isolates were incubated for a certain period of time and stained with iodine solution to determine the size of the transparent ring. .

Through the above measurement of acid production capacity and starch decomposition activity, 36 strains out of 160 strains isolated from grain malt, 11 strains out of 26 strains isolated from grain, and 22 strains out of 144 strains from yeast were screened. Was screened secondary.

Cultivation of Selected Strains

end. Determination of mold spores by medium for optimal solid medium selection

In order to select the optimum solid medium of the fungus, after culturing the fungus using four different media, the result of measuring the spores is shown in Table 1. Strain N159-1 produced 3.65 × 10 ⁷ CFU / mL of spores grown in PDA + Bran medium, more than three times higher than that of PDA medium, and 4.2 times higher than that of PDA + starch. , PDA + sucrose added 4.2 and 5.2 times more than the medium, respectively. N152-1 strains were 2.05 × 10 ⁷ CFU / mL in PDA + Bran medium, about 1.3 times more than in spores of PDA, and C1-5-2-2 strain in PDA + Bran medium. The spore count of 1.65 × 10 ⁸ CFU / mL was about 1.4 times higher than that of PDA medium. The C16-19 strain also showed a spore count of 2.70 × 10 ⁸ CFU / mL in PDA + bran medium, which was about 3.6 times higher than that of PDA medium (0.75 × 10 ⁸ CFU / mL). The strain of CN20.3.1-4 was 3.00 × 10 ⁷ CFU / mL in PDA + Bran medium, more than 3.1 times higher than the number of spores in PDA, and added PDA + starch and PDA + sucrose. About 3.7 and 2.7 times more than one medium, respectively. The CN12.17.1-3 strain showed 3.20 × 10 ⁷ CFU / mL spores in PDA + bran medium, which was about 2.7 and 3.3 times higher than that of PDA + starch and PDA + sucrose. The CN16.3.1-3 strain had a spore count of 3.60 × 10 ⁷ CFU / mL in PDA + wheat bran medium, which was about three times more than that of PDA and PDA + sucrose. The C1-5-2-1 strain showed 2.50 × 10 ⁸ CFU / mL of PDA + wheat bran medium, more than four times higher than the other three media.

As shown in the above results, all the strains used in the experiment showed that the most spores grew in the medium added PDA + bran regardless of the isolate. The reason why the spore count in bran was higher than other media was because the protein, fat and a large amount of minerals contained in the bran helped spore formation by acting as a nutrient.

Number of mold spores by medium for optimal solid medium selection ( ^* : CFU / mL) Strain badge Spore Count ^* Strain badge Spore Count ^* N159-1 PDA 1.20 × 10 ⁷ CN20.3.1-4 PDA 0.95 × 10 ⁷ PDA + starch 0.85 × 10 ⁷ PDA + starch 0.85 × 10 ⁷ PDA + sucrose 0.70 × 10 ⁷ PDA + sucrose 1.10 × 10 ⁷ PDA + Bran 3.65 × 10 ⁷ PDA + Bran 3.00 × 10 ⁷ N152-1 PDA 1.50 x 10 ⁷ CN12.17.1-3 PDA 2.6 × 10 ⁷ PDA + starch 1.40 × 10 ⁷ PDA + starch 1.15 × 10 ⁷ PDA + sucrose 0.95 × 10 ⁷ PDA + sucrose 0.95 × 10 ⁷ PDA + Bran 2.05 × 10 ⁷ PDA + Bran 3.20 × 10 ⁷ C1-5-2-2 PDA 1.15 × 10 ⁸ CN16.3.1-3 PDA 0.95 × 10 ⁷ PDA + starch 0.95 × 10 ⁸ PDA + starch 1.75 × 10 ⁷ PDA + sucrose 1.55 × 10 ⁸ PDA + sucrose 1.15 × 10 ⁷ PDA + Bran 1.65 × 10 ⁸ PDA + Bran 3.60 × 10 ⁷ C16-19 PDA 0.75 × 10 ⁸ C1-5-2-1 PDA 0.55 × 10 ⁸ PDA + starch 1.35 x 10 ⁸ PDA + starch 0.45 × 10 ⁸ PDA + sucrose 1.10 × 10 ⁸ PDA + sucrose 0.60 × 10 ⁸ PDA + Bran 2.70 × 10 ⁸ PDA + Bran 2.50 × 10 ⁸

I. Determination of Fungal Cell Weight by Different Media for Optimal Liquid Media Selection

     In order to select the optimum liquid medium of the fungus, after culturing the fungus using four media having different compositions, the result of measuring the cell mass is shown in Table 2. Among the eight strains used in the experiment, the bacterial masses of the seven strains except CN12.17.1-3 were in the order of bran medium> PSB medium> PDB medium> starch medium, especially in starch medium compared to wheat bran medium. A small amount was shown. N159-1 strains were 1.84g in bran medium, about 12.2 times higher than starch medium and 3.2 times higher than PDB medium. The strains N152-1 and CN20.3.1-4 were 1.68 and 1.94 g in bran medium, respectively, about 12 times higher than those in starch medium, and CN16.3.1-3 and starch in bran medium. The difference in the cell mass of the medium showed the largest difference of 49 times. The CN12.17.1-3 strains were bran medium, PSB medium, starch medium and PDB medium.

As a result, the cell mass of all strains was the highest in the bran medium. The reason why the cell mass in the bran was higher than the other medium is because the protein, fat and the large amount of minerals contained in the bran acted as nutrients.

Cell mass of mold by medium for optimal liquid medium selection ( ^* : g, dry basis) Strain badge Cell weight ^* Strain badge Cell weight ^* N159-1 PDB 0.56 CN20.3.1-4 PDB 0.35 PSB 0.75 PSB 0.70 Starch Badge 0.15 Starch Badge 0.16 Bran Badge 1.84 Bran Badge 1.94 N152-1 PDB 0.35 CN12.17.1-3 PDB 0.15 PSB 0.73 PSB 0.35 Starch Badge 0.14 Starch Badge 0.29 Bran Badge 1.68 Bran Badge 2.38 C1-5-2-2 PDB 0.57 CN16.3.1-3 PDB 0.21 PSB 0.61 PSB 0.36 Starch Badge 0.23 Starch Badge 0.03 Bran Badge 1.72 Bran Badge 1.47 C16-19 PDB 0.76 C1-5-2-1 PDB 0.65 PSB 1.66 PSB 0.81 Starch Badge 0.07 Starch Badge 0.22 Bran Badge 1.72 Bran Badge 1.73

All. Determination of Spores in Different Grains According to Water Content

     18 kinds of grains including glutinous rice, brown rice, glutinous rice black rice, barley, non-glutinous rice, millet, yulmu, buckwheat, black rice, brown rice glutinous rice, tea, rice barley, waxy barley, green beans, wheat, sorghum, red beans, etc. The result of measuring the number of spores is shown in Table 3.

For grains inoculated with strain N159-1, glutinous rice, non-glutinous rice and jum radish had the highest spore counts of 2.10 × 10 ⁸ CFU / mL, 0.84 × 10 ⁸ CFU / mL and 0.75 × 10 ⁸ CFU / mL, respectively, when the water content was 30%. Brown rice, glutinous black rice, barley, millet and buckwheat had spore counts of 1.69 × 10 ⁸ CFU / mL, 1.97 × 10 ⁸ CFU / mL, 0.83 × 10 ⁸ CFU / mL, and 1.38 × 10 ⁸ , respectively, at 20% water content. CFU / mL, 0.60 × 10 ⁸ CFU / mL, showed the most. Especially, in the case of glutinous rice black rice, millet, and buckwheat, the spore count was 20 times higher than the 30% moisture content.In the case of Yulmu, the spore count was 2 when the moisture content was 30% than the 20% moisture content. It was more than twice as high. Black rice inoculated with C16-19 strain showed 1.62 × 10 ⁸ CFU / mL of water content at 30%, more than double the spore count than 20%, and grains inoculated with C1-5-2-2 strain In the case of brown rice glutinous rice, the amount of spores was high when the water content was 30% and the tea was 20% water content. Rice barley inoculated with CN20.3.1-4 strain had 0.46 × 10 ⁸ CFU / mL of spores at 20% water content, which was higher than that at 30% water content. 0.42 × for barley and green beans inoculated with N152-1 strain, respectively. 10 ⁸ CFU / mL and 0.95 × 10 ⁸ CFU / mL showed high spores at 20% water content. Wheat inoculated with CN12.17.1-3 strains had 1.23 × 10 ⁸ CFU / mL of spores at 30% water content, 2.2 times more than at 20% water content, and sorghum and C1-inoculated with CN16.3.1-3 strains. The red beans inoculated with 5-2-1 strain showed 0.42 × 10 ⁸ CFU / mL and 0.95 × 10 ⁸ CFU / mL, respectively, when the water content was 20%.

In the case of bran, the N159-1 strain showed the most active spores when the moisture content was 30% and the other 7 strains were 20%. Spore count was 0.42 × 10 ⁸ CFU / mL, which was the lowest when inoculated with C16-19 strain, and 6 strains including CN16.3.1-3 and C1-5-2-1 were 0.60 × 10 ⁸ ~ 1.00 × 10 ⁸ Values between CFU / mL were shown. The highest number of spores at 30% water content was 1.29 × 10 ⁸ CFU / mL when inoculated with N159-1 strain among 8 strains.

As a result, the number of spores was increased in the case of 20-30% than when the moisture content of the grain inoculated 10%, and it was found that the growth conditions in the grains were changed according to the type of bacteria.

Spore count in each grain according to moisture content ( ^*: CFU / ml) Strain grain Moisture content (%) 10 20 30 N159-1 Glutinous rice 1.39 × 10 ^{8 *} 2.02 × 10 ⁸ 2.10 × 10 ⁸ Brown rice 0.30 × 10 ⁶ 1.69 × 10 ⁸ 1.43 × 10 ⁸ Glutinous rice black rice 0.20 × 10 ⁸ 1.97 × 10 ⁸ 0.68 × 10 ⁸ barley 1.80 × 10 ⁶ 0.83 × 10 ⁸ 0.48 × 10 ⁸ Rice 0.64 × 10 ⁸ 0.78 × 10 ⁸ 0.84 × 10 ⁸ millet 0.22 × 10 ⁸ 1.38 × 10 ⁸ 0.50 × 10 ⁸ Yulmu 0.48 × 10 ⁷ 0.32 × 10 ⁸ 0.75 × 10 ⁸ buckwheat 0.22 × 10 ⁸ 0.60 × 10 ⁸ 0.27 × 10 ⁸ C16-19 Black rice 1.00 × 10 ⁶ 0.79 × 10 ⁸ 1.62 × 10 ⁸ C1-5-2-2 Brown Rice Sticky Rice 0.84 × 10 ⁷ 0.74 × 10 ⁸ 1.15 × 10 ⁸ Tea 0.31 × 10 ⁷ 0.42 × 10 ⁸ 0.23 × 10 ⁸ CN20.3.1-4 Rice barley 0.25 × 10 ⁸ 0.46 × 10 ⁸ 0.27 × 10 ⁸ N152-1 Waxy barley 0.47 × 10 ⁷ 0.47 × 10 ⁸ 0.27 × 10 ⁸ green gram 0.16 × 10 ⁸ 0.46 × 10 ⁸ 0.26 × 10 ⁸ CN12.17.1-3 wheat 0.42 × 10 ⁷ 0.54 × 10 ⁸ 1.23 × 10 ⁸ CN16.3.1-3 Sorghum 0.58 × 10 ⁷ 0.42 × 10 ⁸ 0.33 × 10 ⁸ C1-5-2-1 red bean 0.50 × 10 ⁸ 0.95 × 10 ⁸ 0.85 × 10 ⁸ N159-1 bran
1.15 × 10 ⁷ 0.71 × 10 ⁸ 1.29 × 10 ⁸ C16-19 0.77 × 10 ⁷ 0.42 × 10 ⁸ 0.41 × 10 ⁸ C1-5-2-2 0.33 × 10 ⁸ 0.99 × 10 ⁸ 0.82 × 10 ⁸ CN20.3.1-4 0.71 × 10 ⁷ 0.68 × 10 ⁸ 0.37 × 10 ⁸ N152-1 0.25 × 10 ⁸ 0.63 × 10 ⁸ 0.52 × 10 ⁸ CN12.17.1-3 0.23 × 10 ⁸ 0.80 × 10 ⁸ 0.52 × 10 ⁸ CN16.3.1-3 0.46 × 10 ⁸ 0.74 × 10 ⁸ 0.60 × 10 ⁸ C1-5-2-1 0.32 × 10 ⁸ 0.74 × 10 ⁸ 0.34 × 10 ⁸

Identification of Selected Strains

       Mold isolated from 289 yeast and 179 grains was isolated from ITS1 (CTTGGTCATTTAGAGGAGT).

AA) and ITS4 (TCCTCCGCTTATTGATATGC) primers were identified, and the results are shown in Table 4. Aspergillus spp. Respectively. Aspergillus spp. Was 6 strains, Absidia sp. 3 strains, and Rhizopus sp. 1 strains. Mycocladus sp., N16 strain were Aspergillus tubingensis and N21 strain were Absidia in N3-2 and N254-2 strains that showed high starch degradation activity in starch and grain medium. corymbifera N152-1 and 4 strains showed good starch decomposition ability in starch medium. N159-1 is Aspergillus oryzae , N176-2 for Mycocladus corymbiferus and N245-3 are Absidia Corymbiferus was identified. Aspergillus in 11 strains isolated from cereals spp. was 5 strains and Mucor sp. was a strain, and Aspergillus was a C16-19 strain that had high activity in starch and grain medium. identified as niger .

Identification of strains (only selected strains were shown) strain no. result strain no. result CN16.3.1-3 Aspergillus oryzae N152-1 Aspergillus oryzae CN20.3.1-4 Aspergillus oryzae N159-1 Aspergillus oryzae C1-5-2-2 Aspergillus oryzae C1-5-2-1 Aspergillus oryzae C16-19 Aspergillus niger

Strain Selection During Yeast Manufacturing

end. Primary Screening of Strains

     Using barley, wheat, millet, non-glutinous rice, tea, glutinous barley, yulmu, glutinous rice, brown rice glutinous rice, green beans, sorghum, buckwheat, glutinous rice black rice, black rice, brown rice glutinous rice, rice barley, red beans, etc. Of the 1173 round yeasts (1x10cm) prepared, strains having high saccharogenic ability were selected according to the isolates of each bacterium for each grain to prepare circular yeasts (5x15cm), and the glycosylation power was measured (Table 5). A total of 23 strains were selected. Among them, 12 strains, including CN20.3.1-4 and CN 12.17.1-3, were isolated from grain malt. Strains isolated from grains were 6 strains including C13-10 and C30-5, and strains from Nuruk were 5 strains including N220-1.

     As a result of saccharification, the yeast inoculated with CN12.17.1-3 in wheat showed the highest saccharification ability with 541.2SP, and the yeast inoculated with CN20.3.1-4 in barley and rice barley was 438.0 and 433.8, respectively. SP was high. Non-glutinous rice, green tea, yulmu, glutinous rice, buckwheat and brown rice glutinated rice inoculated with C1-5-2-2 showed saccharification ability between 120 and 270SP. 201.7, 80.0 and 125.5SP, respectively, showed that the degree of growth of bacteria was different according to the inoculated grains. Among the five isolates from yeast, N152-1-inoculated rice malt had the highest saccharification of 405.9SP, and the yeast inoculated with yulmu, glutinous rice and green beans showed a saccharification between 100 and 120SP. The yeast inoculated with N159-1 also showed different glycation potency according to the grains, with the highest saccharification ability of the inoculated yeast in the mail (302SP), and the green tea and brown rice showed 141.8 and 172.9SP, respectively. As the lowest.

     The results of monitoring the temperature and humidity of Nuruk's interior and soup room during the cultivation and drying process are shown in Figure 15. Shown in The internal temperature of yeast increased gradually, peaking at 2 ~ 3 days of culture, and then gradually decreased. The temperature of souproom was the highest at 34.7 on the second day of culture, which was 61.5% RH, and the humidity was kept high due to rainy and snowy weather until 7 days before artificial drying began.

The results of the first strain selection were barley and rice barley CN20.3.1-4, wheat CN12.17.1-3, millet CN23.3.1-3, waxy barley N220-1, non-glutinous rice, tea, yulmu, glutinous rice and brown rice glutinous rice. 5-2-2, brown rice and red beans C1-5-2-1, green beans N152-1, sorghum CN16.3.1-3, buckwheat N159-1, glutinous rice black rice C30-5 and black rice C16-19 Appeared to be suitable.

Glucose Power of Nuruk Prepared for Strain Selection (Finally Selected Strains Only Shown) grain Separation
-Grain yeast Glycation Separation
-grain Glycation barley CN20.3.1-4 438.0 wheat CN12.17.1-3 514.2 millet  N159-1 98.0 Rice C1-5-2-2 264.3 Tea C1-5-2-2 170.2 N159-1 141.8 Waxy barley N220-1 381.4 Yulmu C1-5-2-2 127.5 N152-1 100.7 Glutinous rice C1-5-2-2 246.1 N152-1 121.7 Brown rice C1-5-2-1 201.7 N159-1 172.9 green gram C1-5-2-1 80.0 N152-1 102.3 Sorghum CN16.3.1-3 123.9 buckwheat C1-5-2-2 263.3 N159-1 302.0 Glutinous rice black rice CN20.3.1-4 130.7 Black rice  C16-19 143.2 Brown Rice Sticky Rice CN20.3.1-4 156.3 C1-5-2-2 248.6 Rice barley CN20.3.1-4 433.8 N152-1 405.9 red bean C1-5-2-1 125.5

I. Second Screening of Strains

     Using barley, wheat, millet, non-glutinous rice, tea, glutinous barley, yulmu, glutinous rice, brown rice glutinous rice, green beans, sorghum, buckwheat, glutinous rice black rice, black rice, brown rice glutinous rice, rice barley, red beans, etc. Of the 1173 round yeasts (1 × 10 cm) prepared, three strains having high saccharification ability were selected for each grain to prepare round yeasts (5 × 15 cm) to measure glycation ability (Table 6). Among the selected strains, 8 strains were isolated from grain malt, including CN20.3.1-4, CN30.9.1-1, and 6 strains, such as C13-10, and N171-2, 7 from strains. The strain was a total of 21 strains.

     As a result of the measurement of saccharification power, among rice, glutinous rice, glutinous rice black rice, black rice, brown rice glutinous rice and non-glutinous rice were selected as 1 strain from grains and 2 strains from yeast. N220-1, N159-1, C1-5-2-2, etc. Three strains appeared in common. Glutinous rice and non-glutinous rice showed saccharifying ability of over 150SP in all three selected strains. Among them, N159-1-inoculated yeast had the highest glycation potency, and Glutinous rice black rice was also inoculated with two different strains of yeast inoculated with N159-1. It showed much higher glycosylation ability than leaven. In the black rice, only Nuruk inoculated with C16-1 showed more than 100SP of saccharification ability, and brown rice glutinous rice showed more than 220SP in saccharification capacity of N159-1 and C1-5-2-2. In the case of brown rice, one strain was selected for each isolate, and as with glutinous rice, the highest glycation potency of Nuruk was inoculated with N159-1. Among barleys, barley, which is an outer barley, was selected from 1 strain isolated from grains and 2 strains from yeast. All three strains showed higher glycosylation ability than 350SP. Among them, N159-1 inoculated yeast had the highest glycosylation capacity of 431SP. Rice barley was selected from two strains from grain yeast and one strain from yeast, and all three strains showed high glycosylation ability of 400SP or higher, and the yeast inoculated with N152-1 showed the highest glycation ability of 454.1SP. Two isolates from grain yeast and one strain from yeast were selected, and the inoculation of two isolates from yeast showed more than 400SP. Sorghum strains were selected from 1 strain from grain yeast and 2 strains from yeast. Among the three strains, the glycosylation power of koji inoculated with CN16.3.1-3 isolated from grain yeast was 123.3SP, but overall was less than 150SP. Value was shown. In the case of tea and millet, 1 strain from grain and 2 strains from yeast were selected. Among them, the tea was found to have a higher glycosylation of 185.1SP in yeast inoculated with C1-5-2-2, and the millet was 154.5SP in yeast inoculated with N159-1 among strains isolated from yeast. Showed. All yeast strains were isolated from yeast, and the glycosylation power of yeast inoculated with N159-1 was 176.7SP, which was higher than that of the other two strains. The wheat strains selected from the two strains showed more than 300SP of glycosylation ability, and among them, the glycosylation ability of inoculated CN12.17.1-3 was 402.4SP. In case of buckwheat, two strains from grain yeast and one strain from yeast were selected. Among them, the highest glycosylation of N159-1 inoculated with yeast was 248.0SP. All three strains of mung bean were selected from yeast and showed saccharification ability of less than 100SP. The two strains from grain yeast and red beans from strains were selected according to the type of inoculated strain. Among them, the glycation ability of yeast inoculated with C1-5-2-1 was 158.2SP.

     Nuruk was prepared by inoculating strains having excellent glycosylation capacity for each grain, and monitoring the temperature and humidity of nuruk's interior and soup room during the incubation and drying process. The internal temperature of Nuruk increased rapidly after 1 day of incubation and reached the highest value of 46.5 at 2 days. At this time, the room temperature was 40.5 and the humidity was 90% RH. From the 4th day of culture, both the nuruk's temperature and the room temperature were 30, and gradually decreased until the 7th day of artificial drying.

     Selection of the second strain resulted in glutinous rice, brown rice, glutinous black rice, barley mung bean, non-glutinous rice, millet, yulmu and buckwheat N159-1, brown rice glutinous rice and tea, C1-5-2-2, black rice C16-19, sorghum CN16. 3.1-3, C1-5-2-1 for red beans and CN12.17.1-3 for wheat were found to be suitable.

Glucose Power of Nuruk Prepared for Strain Selection (Finally Selected Strains Only Shown) grain Germ Glycation grain Germ Glycation barley N159-1 431.0 green gram N152-1 80.1 N159-1 95.9 wheat CN12.17.1-3 402.4 Sorghum CN16.3.1-3 123.3 millet N159-1 154.5 buckwheat N159-1 248.0 Rice N159-1 213.5 Glutinous rice black rice N159-1 244.0 C1-5-2-2 199.2 Tea C1-5-2-2 185.1 Black rice C16-19 105.4 Waxy barley N152-1
423.5 Brown Rice Sticky Rice N159-1 227.6 C1-5-2-2 247.1 Yulmu N159-1 176.7 Rice barley N152-1 454.1 CN20.3.1-4 420.2 CN12.17.1-3 404.7 Glutinous rice N159-1 250.1 red bean C1-5-2-1 158.2 N152-1 198.7 C1-5-2-2 164.8 Brown rice N159-1 252.3 C1-5-2-1 151.2

As a result, among the 34 strains selected from the 1st and 2nd stages, 6 strains isolated from grain malt, 6 strains isolated from grain, and 5 strains isolated from malt were commonly selected. For each grain, barley CN20.3.1-4 and N159-1, wheat and buckwheat CN12.17.1-3 and N159-1, rice barley CN20.3.1-4 and N152-1, millet CN23.3.1-3 and N159 -1, green barley is N220-1 and N152-1, non-glutinous rice, yulmu, glutinous rice is C1-5-2-2 and N159-1, green beans are N152-1 and N159-1, brown rice is C1-5-2-1 N159-1 glutinous rice black rice is C30-5 and N159-1 brown rice and brown rice glutinous rice C1-5-2-2, red beans C1-5-2-1, sorghum CN16.3.1-3, black rice C16-19 And the captain showed that N159-1 was the best.

Fermented Wine Preparation and Final Strain Selection

     Each of the grains were inoculated with grain yeast, grains, and yeast inoculated with the strains (high strains of saccharification for each grain among CN, C, and N strains in Table 6). The analysis results are shown in Tables 7 and 8.

     The alcohol content was 5.0 ~ 11.7%, pH was 3.87 ~ 5.08, the sugar content was 4.6 ~ 11.7 ° Brix, the acidity was 0.11 ~ 0.66, the reducing sugar was 0.52 ~ 4.50mg / mL, and the amino acid was below 0.73g / 100mL. The alcohol content of the yeast inoculated with N159-1 strain in glutinous rice was 10.9%, the sugar content was 9.3, the reducing sugar was 4.35mg / mL, and the acidity was 0.11. The alcohol content of sake made with malt inoculated with brown rice, glutinous rice black rice, barley, and millet N159-1 was 10.0, 9.9, 9.7, and 9.8%, respectively, and the acidity was 0.14, 0.19, 0.14, 0.19, respectively. . The alcohol content of Nuruk inoculated with C16-19 strain in black rice was 9.0%, reducing sugar was 1.61mg / mL, and acidity was 0.66. Nuruk wines inoculated with C1-5-2-2 strains in brown rice glutinous rice and tea were lower in alcohol content of 8.3 and 7.9, respectively, with acidity of 0.22, 0.61 and reducing sugar of 1.61 and 0.57 mg / mL, respectively. appear. The alcohol using Nuruk inoculated with N152-1 strains in rice barley and waxy barley showed alcohol content of 9.9, 9.1%, and acidity of 0.16 and 0.13, respectively, and the reducing sugars of Nuruk inoculated with N152-1 strains. Alcohol was the highest at 4.50 mg / mL. Nuruk inoculated with strain N159-1 in Yulmu and buckwheat, respectively, had a low alcohol content of 8.1 and 7.2%, respectively, and the acidity was 0.52 and 0.49, respectively. . Nuruk, inoculated with CN12.17.1-3 strains, showed 9.5% alcohol content and 1.62 mg / mL of reducing sugar, and the highest sugar content was 11.7 ° Brix and 0.73g / 100mL of amino acid. The alcohol content of Nuruk inoculated with CN16.3.1-3 strains was the lowest at 5.0%, and the lowest sugar content was 4.6 ° Brix. Nuruk inoculated with N159-1 strain in non-glutinous rice showed the highest alcohol content (11.7%) and low acidity (0.12).

     Organic acids and free sugars were analyzed by citric acid, malic acid, succinic acid, lactic acid and acetic acid. There were three free sugars: fructose, glucose and maltose. In the case of malt inoculated with glutinous rice inoculated with N159-1 strain, succinic acid showed the highest content of 9.91mg / mL among lactic acid, and lactic acid, malic acid and citric acid were between 0.01 ~ 0.54mg / mL. In the two free sugars detected, the content of glucose was the highest at 1.82mg / mL. In the yeast of inoculated yeast seeded with N159-1 in brown rice, organic acid had the highest succinic acid of 6.82 mg / mL, and lactic acid and citric acid of 1.31 and 0.26 mg / mL, respectively, and free sugar of maltose. The content was 0.85 mg / mL, indicating a high content. The organic acid of liquor made from N159-1 strain and yeast made from glutinous rice black rice was 1.37 and 2.94mg / mL, respectively, and the free sugar contained maltose 0.75mg / mL, respectively. The organic acid of Yakju made from yeast inoculated with C16-19 strain in black rice showed 20.56mg / mL of lactic acid, which was much higher than other organic acids, and the free sugars of three kinds were between 0.19 ~ 1.01mg / mL. Indicated. Yeoju prepared with Nuruk inoculated with C1-5-2-2 strain in brown rice glutinous rice showed the highest content of lactic acid (4.89mg / mL) among the organic acids, and the other two organic acids had a value below 0.50mg / mL. Free sugar had the highest maltose content of 0.43mg / mL. The wine made with Nuruk inoculated with barley N159-1 strain showed the highest content of succinic acid (3.61mg / mL) among the organic acids and less than 0.35mg / mL of the three free sugars. The organic acid of Yakju made from Nuruk inoculated with N152-1 strains of rice barley showed the highest value of 6.49mg / mL in succinic acid, and fructose and glucose of 0.17 and 0.35mg / mL in free sugar, respectively. The organic acid of the liquor made from the barley and N152-1 strains showed 3.95 and 1.75mg / mL of succinic acid and lactic acid, respectively. The fructose and glucose of 0.42 and 0.73mg / mL of fructose and glucose were free sugar, respectively. Indicated. The organic acid of the liquor made from Nuruk was inoculated with C1-5-2-2 and the highest was lactic acid (17.75mg / mL), and the free sugar showed maltose (0.50mg / mL). In the yeast made with inoculated with N159-1 strain in the millet, succinic acid and lactic acid showed 3.18 and 2.17 mg / mL, respectively, and free sugar of fructose was 0.14 mg / mL. The organic acid of Yakju made from yeast inoculated with Yulmu inoculated with N159-1 strain showed the highest content of lactic acid (16.12mg / mL), and the other two organic acids showed values between 0.78 ~ 1.47mg / mL. The fructose and glucose were 0.15 and 0.27mg / mL, respectively. In medicinal herb made from yeast made from wheat and CN12.17.1-3 strain, organic acid showed high value of succinic acid (7.56mg / mL) and three free sugars between 0.23 ~ 0.93mg / mL. . In the yeast liquor prepared by inoculating buckwheat with N159-1 strain, the organic acid had the highest lactic acid content of 12.92mg / mL and the free sugar showed maltose of 1.42mg / mL. The liquor made with the yeast inoculated with sorghum CN16.3.1-3 had the highest lactic acid content of 7.08mg / mL in the organic acid, and the other two types of organic acids were detected below 1.75mg / mL. , Free sugar showed high maltose of 0.71mg / mL. In the yeast produced by inoculation with non-rice inoculated with N159-1 strain, organic acid showed the highest succinic acid with 4.49mg / mL and glucose with 1.97mg / mL with high sugar.

     Sensory evaluation (Table 9) showed significant differences in terms of appearance, taste and overall preference (p <0.01). The alcohol using Nuruk prepared by inoculating glutinous rice and brown rice with N159-1 strain respectively showed more than 7.00 and 5.50 in appearance and flavor, respectively, but the taste and overall acceptability were below 5.00. The liquor made with yeast inoculated with N159-1 strain in glutinous rice black rice showed 7.00, 6.00, and 5.00 tastes, and 5.33, respectively. Appearance of liquor using yeast inoculated with C16-19 strain and C1-5-2-2 strain in black and brown rice glutinous rice respectively was 7.33, but taste and overall acceptability were low. The wine made with yeast inoculated with barley N159-1 strain showed the appearance, aroma and taste of 6.33, 6.00 and 5.00, respectively, and the overall acceptability was 5.00. The appearance and aroma of alcohol using Nuruk inoculated with N152-1 strain in rice barley were 7.00 and 5.67, respectively. The appearance, aroma and taste of alcohol using Nuruk inoculated with N152-1 strain in rice barley 7.33, 6.67, and 5.00, respectively, and the overall acceptability was 5.33. The appearance, aroma and taste of liquor made from Nuruk inoculated with N159-1 strains were 6.67, 6.33, and 5.67, respectively. Liquor made from yeast inoculated with C1-5-2-2 strains in Chajo, Yulmu, and wheat, yeast inoculated with N159-1 strains in Yulmu, and yeast inoculated with CN12.17.1-3 strains in wheat. The degree of preference was higher than usual, but was low in taste and overall preference. The liquor made with malt inoculated with N159-1 strain in non-rice was 7.00, 5.67 and 6.00 in appearance, aroma, and taste, respectively, with the highest palatability of 6.00. On the other hand, the preference of liquor made with buckwheat and sorghum was very low overall. Overall, liquor made from malt inoculated with N159-1 or N152-1 strains of non-glutinous rice, millet, waxy barley, glutinous rice black rice and barley was found to have high preference.

Analysis of liquor dipped using non-glutin and starch as a raw material inoculated by inoculating strains isolated from grain koji, grains, and yeast (strains with high saccharifying ability for each grain among the CN, C, and N strains in Table 6) The results are shown in Tables 10 and 11.

     The alcohol content was 7.9 ~ 15.8%, pH was 4.48 ~ 5.44, the sugar content was 6.1 ~ 11.6 ° Brix, the acidity was 0.08 ~ 0.19, the reducing sugar was 1.33 ~ 13.74 mg / mL, and the amino acid was 0.12 ~ 0.49 mg / 100mL. The alcohol content of Nuruk inoculated with N159-1 strain in glutinous rice, brown rice, and glutinous rice black rice, respectively, was relatively high (12.2 ~ 12.6%), and the acidity was 0.11 ~ 0.12, reducing sugar was 5.42 ~ 6.14 mg / mL. Nuruk inoculated with C16-19 strain in black rice had the highest acidity of 0.19 and the lowest reducing sugar of 1.33 mg / mL. Of yeast inoculated with strain N152-1 in yeast, yeast inoculated with strain C1-5-2-2, yeast inoculated with strain N159-1 in Yulmu, and yeast inoculated with strain N159-1 in buckwheat The alcohol content was relatively high with 12.1 ~ 12.6%, acidity of 0.12 ~ 0.17, and sugar content of 10.5 ~ 11.6 ° Brix. The reduced sugar content of Nuruk inoculated with N152-1 strain in rice barley was 13.74 mg / mL. Was the lowest. Yeast inoculated with C1-5-2-2 strain in brown rice, yeast inoculated with N159-1 strain in barley, yeast inoculated with N159-1 strain in millet, and yeast inoculated with CN12.17.1-3 strain in wheat The alcohol content was 11.2, 11.2, 11.4, 11.9%, respectively, and the acidity was 0.13, 0.14, 0.14, 0.13, respectively. The alcohol content of Nuruk inoculated with N159-1 strain in mung beans was low as 9.9% and the lowest in acidity was 0.08. The alcohol in Nuruk inoculated with C1-5-2-1 strain in red beans was alcohol. The highest content was 15.8%. Overall, fermentation of sorghum and mung beans was slow.

     Organic acids and free sugars were analyzed by citric acid, malic acid, succinic acid and lactic acid. There were three free sugars, fructose, glucose and maltose. In the case of malt inoculated with glutinous rice inoculated with N159-1 strain, succinic acid showed the highest content of 11.56mg / mL among lactic acid, and lactic acid, malic acid and citric acid were between 0.10 ~ 0.66mg / mL. Glucose content of the two detected free sugars was the highest at 2.97mg / mL. In the yeast of inoculated yeast inoculated with strain N159-1 in brown rice, organic acid had the highest succinic acid (13.76 mg / mL) and free sugar had the highest content of glucose (2.17 mg / mL). Free sugar showed a value of 0.20 mg / mL or less. The organic acid of liquor made from the yeast made with strain N159-1 and glutinous rice black rice showed 17.09mg / mL of succinic acid, and the other three organic acids showed below 0.30mg / mL. High content was found at 3.00 mg / mL. The organic acid of Yakju made from yeast inoculated with C16-19 strain in black rice showed 7.35mg / mL of succinic acid, which was much higher than other organic acids. Indicated. Yeoju prepared with Nuruk inoculated with C1-5-2-2 strain in brown rice glutinous rice showed the highest content of succinic acid (14.96mg / mL) among organic acids and glucose (1.20mg / mL). In the wine made with Nuruk inoculated with barley N159-1 strain, succinic acid was 21.91mg / mL, and the two free sugars were 0.16 and 2.32mg / mL, respectively. The organic acid of Yakju made from yeast inoculated with N152-1 strains of barley showed the highest succinic acid level of 23.80mg / mL and glucose of 6.25mg / mL in free sugar. The organic acid of liquor made from the barley and N152-1 strain showed succinic acid value of 12.44mg / mL, and the free sugar showed 3.27mg / mL of glucose. The organic acid of the liquor made by inoculating C1-5-2-2 strain in the tea was highest in succinic acid (21.73mg / mL), and the value of three free sugars was between 0.08 ~ 1.00mg / mL. In the yeast made with inoculated with N159-1 strain in the millet, succinic acid and lactic acid were 12.97 and 2.33 mg / mL, respectively, and free sugar was 2.83 mg / mL, respectively. The organic acid of Yakju made from yeast inoculated with Yulmu inoculated with N159-1 strain showed 25.41mg / mL of succinic acid, which was much higher than other organic acids. The content of / mL is shown. In medicinal herbs made from yeast made from wheat and CN12.17.1-3 strain, organic acid showed high value of succinic acid (7.80mg / mL) and two free sugars between 0.20 ~ 2.67mg / mL. . In the alcoholic beverage made by inoculating buckwheat with N159-1 strain, succinic acid and lactic acid showed 11.52 and 2.18mg / mL, respectively, and free sugar showed 2.22mg / mL of glucose. . The liquor made with the yeast inoculated with sorghum CN16.3.1-3 had the highest succinic acid content of 8.98 mg / mL among the organic acids, and the other two organic acids were detected in amounts less than 1.10 mg / mL. The three free sugars showed values less than 1.00 mg / mL. Citric acid, succinic acid, and succinic acid were found to be 1.21 and 1.74 mg / mL, respectively, and the organic acids of yeast made from Nuruk inoculated with N159-1 strain of mung beans were 0.95 mg / mL or less. The content of was shown. The organic acid of Yakju prepared by using yeast inoculated with red bean C1-5-2-1 strain showed high succinic acid of 3.34mg / mL, and free sugar of glucose and maltose of 2.19 and 0.10mg / mL, respectively. Showed. In the yeast produced by inoculation with non-rice inoculated with N159-1 strain, organic acid showed the highest succinic acid with 4.49mg / mL and glucose with 1.97mg / mL with high sugar.

     Sensory test results (Table 12) Sensory test results of liquor using yeast prepared by inoculating N159-1 strain on glutinous rice showed 7.33 and 7.00 appearance and fragrance preference, respectively, and overall acceptability was 5.67. Nuruk was inoculated with a liquor of 6.00 or more in all items. The appearance, aroma and taste of Nuruk inoculated with glutinous rice black rice inoculated with N159-1 strain were 7.00, 7.00, and 6.33, respectively, and the overall acceptability was 6.67. In the malt inoculated with black rice C16-19 strain, All items showed more than 6.00 degree of preference. The Nuruk wine, which was inoculated with C1-5-2-2 strain in brown rice glutinous rice, had a high appearance, aroma, and taste of 7.67, 7.33, and 7.33, respectively. The overall acceptability of Nuruk inoculated with N159-1 strain to barley, Nuruk inoculated with N152-1 strain to rice barley, and liquor inoculated with N152-1 strain to barley were 5.67, 4.67, and 6.67, respectively. . The appearance and aroma of Nuruk inoculated with C1-5-2-2, N159-1, and N159-1 strains in the tea, millet, and yulmu, respectively, was over 5.50, but the taste and overall acceptability were less than 5.00. . The appearance and fragrance of liquor made from malt inoculated with CN12.17.1-3 strain in wheat and malt inoculated with CN16.3.1-3 strain in sorghum showed more than 5.50 and overall acceptability was 5.00, respectively. Nuruk, which was inoculated with N159-1 strain in non-glutinous rice, had the appearance, aroma, and taste of 7.00, 5.57, and 6.00, respectively. The appearance of liquor inoculated with buckwheat, mung beans, and red beans was 5.50 or more, but the aroma, taste, and overall acceptability were low. Overall, liquor made with malt inoculated with brown rice, glutinous rice black rice, glutinous barley, brown rice, black rice, non-glutinous rice, glutinous rice, and barley showed high acceptability. All items showed significant difference (p <0.05, 0.001, 0.0001).

The fermentation pattern, characteristics, and preference of fermented Yakju using inoculum inoculated with strains with excellent saccharification ability for each grain were selected, considering the saccharification ability of each grain malt.

Characterization of Yakju Using Yeast Inoculated with Highly Saccharified Bacteria (Starch Raw Material: Grain) grain Strain Alc (%) pH Acidity Sugar content
(° Brix) Reducing sugar
(mg / mL) amino acid
(g / 100mL) Glutinous rice N159-1 10.9 4.62 0.11 9.3 4.35 0.10 Brown rice N159-1 10.0 5.08 0.14 8.4 1.79 0.12 Glutinous rice black rice N159-1 9.9 4.82 0.19 7.9 0.64 0.14 Black rice C16-19 9.0 3.87 0.66 8.7 1.61 0.31 Brown Rice Sticky Rice C1-5-2-2 8.3 4.61 0.22 6.9 0.58 0.11 barley N159-1 9.7 4.28 0.14 8.5 3.41 0.04 Rice barley N152-1 9.9 4.54 0.16 9.1 2.93 0.11 Waxy barley N152-1 9.1 4.54 0.13 8.7 4.50 0.13 Tea C1-5-2-2 7.9 3.88 0.61 7.6 0.57 0.26 millet N159-1 9.8 4.90 0.19 8.7 0.68 0.29 Yulmu N159-1 8.1 4.05 0.52 8.0 0.52 0.27 wheat CN12.17.1-3 9.5 4.89 0.26 11.7 1.62 0.73 buckwheat N159-1 7.2 4.37 0.49 7.9 0.95 0.35 Sorghum CN16.3.1-3 5.0 4.08 0.35 4.6 0.59 0.00 Rice N159-1 11.7 4.40 0.12 9.6 4.48 0.13

Characterization of Yakju Using Yeast Inoculated with Highly Saccharified Bacteria (Starch Raw Material: Grain) grain Strain Organic acid (mg.mL) Free sugar (mg / mL) citric malic succinic lactic acetic fructose glucose maltose Glutinous rice N159-1 0.16 0.01 9.21 0.54 - - 1.82 0.17 Brown rice N159-1 0.26 - 6.82 1.31 - 0.10 0.36 0.85 Glutinous rice black rice N159-1 0.40 - 1.37 2.94 - 0.17 0.27 0.75 Black rice C16-19 0.21 - 0.17 20.56 1.94 0.19 0.82 1.01 Brown Rice Sticky Rice C1-5-2-2 0.37 - 0.43 4.89 - 0.13 0.24 0.43 barley N159-1 0.04 0.01 3.61 0.71 - 0.15 0.33 0.30 Rice barley N152-1 - 0.01 6.49 0.78 - 0.17 0.35 - Waxy barley N152-1 - - 3.95 1.75 - 0.42 0.73 - Tea C1-5-2-2 - - 3.68 17.75 0.71 0.13 - 0.50 millet N159-1 0.29 - 3.18 2.17 - 0.14 - - Yulmu N159-1 - - 1.47 16.12 0.78 0.15 0.27 - wheat CN12.17.1-3 - - 7.56 2.96 - 0.23 0.23 0.93 buckwheat N159-1 - - 1.49 12.92 1.81 0.09 0.23 1.42 Sorghum CN16.3.1-3 - - 1.74 7.08 0.46 0.09 0.42 0.71 Rice N159-1 0.05 - 4.49 0.58 - 0.13 1.97 -

Characterization of Yakju Using Yeast Inoculated with Highly Saccharified Bacteria (Starch Raw Material: Grain) grain Strain Exterior Symbol ^** Scent Taste sign ^** Overall preference ^** Glutinous rice N159-1 7.33 ^ab 5.67 ^abc 4.33 ^abc 4.67 ^abc Brown rice N159-1 7.67 ^a 5.67 ^abc 4.67 ^abc 4.67 ^abc Glutinous rice black rice N159-1 7.00 ^ab 6.00 ^ab 5.00 ^ab 5.33 ^ab Black rice C16-19 7.33 ^ab 4.00 ^bc 3.67 ^bcde 4.00 ^abcd Brown Rice Sticky Rice C1-5-2-2 7.33 ^ab 6.67 ^a 4.00 ^abcd 4.00 ^abcd barley N159-1 6.33 ^abc 6.00 ^ab 5.00 ^ab 5.00 ^abc Rice barley N152-1 7.00 ^ab 5.67 ^abc 3.67 ^bcde 3.67 ^bcde Waxy barley N152-1 7.33 ^ab 6.67 ^a 5.00 ^ab 5.33 ^ab Tea C1-5-2-2 6.67 ^abc 5.33 ^abc 2.67 ^cdef 3.00 ^cde millet N159-1 6.67 ^abc 6.33 ^a 5.67 ^ab 5.67 ^ab Yulmu N159-1 7.00 ^ab 6.33 ^a 3.67 ^bcde 3.67 ^bcde wheat CN12.17.1-3 5.00 ^cd 5.33 ^abc 2.00 ^def 2.00 ^de buckwheat N159-1 3.67 ^d 3.67 ^c 1.67 ^ef 1.67 ^e Sorghum CN16.3.1-3 5.67 ^bc 4.00 ^bc 1.33 ^f 1.67 ^e Rice N159-1 7.00 ^ab 5.67 ^abc 6.00 ^a 6.00 ^a LSD (5%) 1.75 ns 2.23 2.08

Characteristics of Yakju Using Nuruk Inoculated with Bacteria with Excellent Saccharification Power (Starch Raw Material: Non-glutinous Rice) grain Strain Alc (%) pH Acidity Sugar content
(° Brix) Reducing sugar
(mg / mL) amino acid
(g / 100mL) Glutinous rice N159-1 12.5 4.85 0.11 10.3 5.42 0.26 Brown rice N159-1 12.6 4.96 0.12 11.2 6.14 0.26 Glutinous rice black rice N159-1 12.2 4.87 0.11 10.7 6.10 0.26 Black rice C16-19 11.5 4.48 0.19 9.1 1.33 0.20 Brown Rice Sticky Rice C1-5-2-2 11.2 4.71 0.13 10.2 3.23 0.19 barley N159-1 11.2 4.52 0.14 11.0 8.94 0.26 Rice barley N152-1 10.7 4.75 0.11 11.1 13.74 0.36 Waxy barley N152-1 12.4 4.65 0.12 11.0 9.58 0.30 Tea C1-5-2-2 12.6 4.69 0.13 10.5 3.99 0.24 millet N159-1 11.4 4.64 0.14 10.0 5.83 0.30 Yulmu N159-1 12.2 4.93 0.14 11.6 9.74 0.49 wheat CN12.17.1-3 11.9 4.69 0.13 10.9 9.03 0.31 buckwheat N159-1 12.1 4.61 0.17 10.9 4.34 0.29 Sorghum CN16.3.1-3 7.9 4.48 0.12 6.1 2.03 0.12 green gram N159-1 9.9 5.44 0.08 8.1 6.03 0.13 red bean C1-5-2-1 15.8 5.11 0.14 10.5 11.65 0.13

Characteristics of Yakju Using Nuruk Inoculated with Bacteria with Excellent Saccharification Power (Starch Raw Material: Non-glutinous Rice) grain Strain Organic acid (mg.mL) Free sugar (mg / mL) citric malic succinic lactic fructose glucose sucrose Glutinous rice N159-1 0.10 0.17 11.56 0.66 - 2.97 0.18 Brown rice N159-1 0.25 0.23 13.76 0.26 0.07 2.17 0.16 Glutinous rice black rice N159-1 0.23 0.27 17.09 0.29 0.09 3.00 0.20 Black rice C16-19 0.51 - 7.35 0.58 0.35 0.28 0.14 Brown Rice Sticky Rice C1-5-2-2 0.23 0.25 14.96 0.39 - 1.20 0.14 barley N159-1 0.07 0.37 21.91 0.85 - 2.32 0.16 Rice barley N152-1 0.12 0.30 23.80 0.58 0.06 6.25 0.22 Waxy barley N152-1 0.08 0.27 12.44 0.69 0.07 3.28 0.22 Tea C1-5-2-2 0.27 0.09 21.73 1.06 0.08 1.00 0.19 millet N159-1 0.17 - 12.97 2.33 0.11 2.83 0.18 Yulmu N159-1 0.52 0.31 25.41 0.56 0.10 3.83 0.45 wheat CN12.17.1-3 0.05 0.24 7.80 0.49 - 2.67 0.20 buckwheat N159-1 - 0.15 11.52 2.18 - 2.22 0.23 Sorghum CN16.3.1-3 0.27 - 8.98 1.05 0.06 0.98 0.13 green gram N159-1 1.21 0.11 1.74 0.71 0.12 0.95 0.09 red bean C1-5-2-1 0.91 0.21 3.34 0.91 - 2.19 0.10 Rice N159-1 0.05 - 4.49 0.58 0.13 1.97 -

Characteristics of Yakju Using Nuruk Inoculated with Bacteria with Excellent Saccharification Power (Starch Raw Material: Non-glutinous Rice) grain Strain Appearance Symbol ^* Fragrance Hodo ^*** Taste sign ^**** Overall preference degree ^**** Glutinous rice N159-1 7.33 ^ab 7.00 ^ab 5.33 ^bcd 5.67 ^bcd Brown rice N159-1 6.67 ^b 6.67 ^ab 6.00 ^abc 6.00 ^abc Glutinous rice black rice N159-1 7.00 ^ab 7.00 ^ab 6.33 ^ab 6.67 ^ab Black rice C16-19 6.67 ^b 6.00 ^ab 6.00 ^abc 6.00 ^abc Brown Rice Sticky Rice C1-5-2-2 7.67 ^a 7.33 ^a 7.33 ^a 7.33 ^a barley N159-1 7.00 ^ab 7.00 ^ab 5.67 ^bcd 5.67 ^bcd Rice barley N152-1 7.00 ^ab 7.00 ^ab 4.67 ^cde 4.67 ^cde Waxy barley N152-1 7.00 ^ab 7.00 ^ab 6.67 ^ab 6.67 ^ab Tea C1-5-2-2 7.00 ^ab 6.33 ^ab 4.33 ^def 4.33 ^def millet N159-1 7.33 ^ab 5.57 ^bc 3.67 ^ef 3.67 ^ef Yulmu N159-1 7.00 ^ab 6.67 ^ab 4.67 ^cde 4.67 ^cde wheat CN12.17.1-3 6.67 ^b 6.00 ^ab 4.67 ^cde 5.00 ^cde buckwheat N159-1 6.67 ^b 4.33 ^cd 3.67 ^ef 3.67 ^ef Sorghum CN16.3.1-3 6.67 ^b 5.57 ^bc 5.33 ^bcd 5.00 ^cde green gram N159-1 6.67 ^b 3.67 ^d 3.00 ^f 3.00 ^f red bean C1-5-2-1 5.67 ^c 4.00 ^d 3.67 ^ef 3.67 ^ef Rice N159-1 7.00 ^ab 5.57 ^bc 6.00 ^abc 6.00 ^abc LSD (5%) 0.84 1.60 1.56 1.45

Based on the saccharification power of the yeast produced by each grain and the result of analyzing and using the yeast, the strain was selected for the preparation of the dedicated yeast for each grain (Table 13). Finally, 8 strains were selected as Aspergillus 1 strain of flavus, 1 strain of Aspergillus niger and Aspergillus 6 strains were oryzae . The strains isolated from grain koji were CN20.3.1-4, CN16.3.1-3 and CN12.17.1-3, which were found to be suitable for the production of dedicated yeast for rice barley, sorghum and wheat, respectively. The selection criteria for rice barley was 6.25, which was the preference of Yakju prepared using rice barley malt, and sorghum and wheat showed higher glycosylation ability of malt prepared by inoculating the selected bacteria. The strains isolated from grains were C16-19, C1-5-2-2, and C1-5-2-1, which were found to be suitable for the production of exclusive malt of black rice, brown rice glutinous rice and tea, and red beans. The selection criteria were that black rice and brown rice glutinous rice had high saccharification power, and the preference score of Yakju made by using non-glutinous rice as starch material was 6.67, and tea and red beans were C1-5-2-2 and This is because the saccharification power of Nuruk prepared by inoculation with C1-5-2-1 was higher than that of Nuruk using another strain. The strains isolated from Nuruk were N159-1 and N152-1, and N159-1 was found to be suitable for the production of exclusive yeast of glutinous rice, brown rice, glutinous black rice, barley, non-glutinous rice, millet, yulmu and buckwheat. It has been shown to be suitable for the preparation of dedicated malt of mung beans. The selection criteria were glutinous rice, brown rice, glutinous rice black rice, barley, waxy barley, and non-glutinous rice, which had high saccharification ability, and the preference score of Yakju made by using non-glutinous rice as starch raw material was 6.00 or higher. Mung bean and buckwheat had higher glycosylation ability of yeast prepared by inoculating each of the selected strains.

Strains for the manufacture of exclusive yeast for each grain No. grain Strain No. grain Strain One Glutinous Rice ^* N159-1 (Accession Number: KCTC11927BP) 10 Red beans ^** C1-5-2-1 (Accession No .: KCTC11921BP) 2 Brown Rice ^* N159-1 (Accession Number: KCTC11927BP) 11 Rice ^* N159-1 (Accession Number: KCTC11927BP) 3 Glutinous rice black rice ^* N159-1 (Accession Number: KCTC11927BP) 12 Chajo ^** C1-5-2-2 (Accession No .: KCTC11922BP) 4 Black rice ^* C16-19 (Accession No .: KCTC11923BP) 13 Millet ^** N159-1 (Accession Number: KCTC11927BP) 5 Brown Rice Sticky Rice ^* C1-5-2-2 (Accession No .: KCTC11922BP) 14 Admiral ^** N159-1 (Accession Number: KCTC11927BP) 6 Barley ^* N159-1 (Accession Number: KCTC11927BP) 15 Wheat ^** CN12.17.1-3 7 Rice Barley ^*** CN20.3.1-4 (Accession Number: KCTC11925BP) 16 Mung Bean ^** N152-1 (Accession No .: KCTC11926BP) 8 Waxley ^* N152-1 (Accession No .: KCTC11926BP) 17 Buckwheat ^** N159-1 (Accession Number: KCTC11927BP) 9 Sorghum ^** CN16.3.1-3 (accession number:
KCTC11924BP)

*: Excellent glycation ability + preference

**: Excellent glycation power

***: Excellent sign

Physiological Functionality Measurement of Fermented Wine Using Selected Nuruk

     Antihypertensive angiotensin converting enzyme (ACE) inhibitory activity was measured by using acetone extracted from rabbit lung with a commercial standard ACE and Hip-His-Leu as a major physiological function. DPPH method, SOD-like activity was measured by absorbance measurement using pyrogallol, and anti-dementia acetylcholine esterase inhibitory activity was measured by Ellmen method based on acetylcholin (Table 14).

     Finally, after preparing traditional folk liquor using the selected yeast, their physiological functionalities were examined. 4 The traditional liquor made with yeast was the highest with 90.5%.

     Antioxidant activity, which inhibits aging by preventing oxidation of organic substances in the body, was also highest in traditional liquor made with No 4 Nuruk (25.3%).

There was no or very weak SOD-like activity related to the inhibitory activity of acetylcholinesterase (AChE), which esterifies the neurotransmitter acetylcholine, causing dementia, thrombolytic activity, and anti-aging.

Physiological Functionality of Alcoholic Beverages Prepared with Selected Nuruks No. grain Antioxidant activity (%) SOD-like activity (%) ACE inhibitory activity (%) ¹⁾ Fibrinolytic activity
(clearzone: mm) AChE inhibitory activity (%) ¹⁾ 2 Brown rice 9.6 (± 0.8) n.d 81.9 (± 0.6) n.d - 3 Glutinous rice black rice n.d n.d 87.9 (± 0.3) n.d - 4 Black rice 25.3 (± 1.1) n.d 90.5 (± 0.7) n.d - 5 Brown Rice Sticky Rice 22.1 (± 0.6) 3.76 (± 0.26) 77.2 (± 1.0) n.d - 8 Waxy barley 7.4 (± 0.4) n.d 77.6 (± 0.8) n.d - 11 Rice 19.2 (± 1.1) n.d 80.7 (± 0.1) n.d - 12 Tea 7.5 (± 0.3) n.d 72.3 (± 0.4) n.d -

1) ACE; angiotensin I-converting enzyme, AChE; acetylcholinesterase.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Korea Biotechnology Research Institute KCTC11922 20110426

Claims (17)

A method of producing yeast by inoculating brown rice glutinous rice as inoculated with a strain of Aspergillus duckase C1-5-2-2 (Aspergillus oryzae, Microorganism Accession No .: KCTC11922BP).
The method of claim 1,
The strain is a method for producing yeast, characterized in that cultured in a solid medium consisting of potato medium (PDA, Potato Dextrose Agar) and bran.
The method of claim 1,
The strain is a method for producing yeast, characterized in that cultured in a liquid medium of bran.
The method of claim 1,
The strain is a method of producing yeast, characterized in that cultured in a medium of 20 to 30% by weight of the brown rice glutinous rice.
Nuruk prepared by inoculating brown rice glutinous rice inoculated with Aspergillus duckase C1-5-2-2 (Aspergillus oryzae, microorganism accession number: KCTC11922BP).
6. The method of claim 5,
The strain is yeast, characterized in that cultured in a solid medium consisting of potato medium (PDA, Potato Dextrose Agar) and bran.
6. The method of claim 5,
The strain is yeast, characterized in that cultured in a liquid medium of bran.
6. The method of claim 5,
The strain is yeast, characterized in that cultured in a medium of 20 to 30% by weight of the brown rice glutinous rice.
A method of preparing fermented liquor by inoculating brown rice glutinous rice by inoculation with Aspergillus aurise C1-5-2-2 (Aspergillus oryzae, Microorganism Accession No .: KCTC11922BP) cultured as a strain.
The method of claim 9,
The strain is a method for producing a fermentation strain, characterized in that cultured in a solid medium consisting of potato medium (PDA, Potato Dextrose Agar) and bran.
The method of claim 9,
The strain is a method for producing a fermented liquor characterized in that cultured in a liquid medium of bran.
The method of claim 9,
The strain is a method of producing a fermented liquor characterized in that the culture of the brown rice glutinous rice in the content of 20 to 30% by weight.
A fermented liquor fermented with yeast prepared by inoculating Brown Rice Glutinous Rice as a strain inoculated with Aspergillus duckase C1-5-2-2 (Aspergillus oryzae, Microorganism Accession No .: KCTC11922BP).
The method of claim 13,
The strain is fermented liquor, characterized in that cultured in a solid medium consisting of potato medium (PDA, Potato Dextrose Agar) and bran.
The method of claim 13,
The strain is fermented liquor, characterized in that cultured in a liquid medium of bran.
The method of claim 13,
The strain is fermented liquor, characterized in that cultured in a medium of 20 to 30% by weight of the brown rice glutinous rice.
The method of claim 13,
The fermented wine is any one selected from the group consisting of Takju, Yakju, Cheongju, beer and fruit wine.