KR20150052555A - Flavour promoted distilled spirit using yeast saccharomyces cerevisiae 88-4 - Google Patents

Abstract

The present invention relates to a novel saccharomyces cerevisiae strain used in manufacturing a distilled spirit, a distilled spirit manufactured by using the same and a distilled spirit manufacturing method using the same. The distilled spirit manufactured according to the present invention has a very excellent characteristic of an organic function because of proper balance among flavor contribution ingredients by the new saccharomyces cerevisiae 88-4 strain when alcohol fermentation is performed.

Description

FLAVOR PROMOTED DISTILLED SPIRIT USING YEAST SACCHAROMYCES CEREVISIAE 88-4 < / RTI > prepared by using the yeast Saccharomyces cerevisiae 88-4,

The present invention relates to novel saccharomyces cerevisiae ( Saccharomyces < RTI ID = 0.0 > cerevisiae strain, a distillation column manufactured using the same, and a method for producing a distillation column using the same.

In general, alcohol means a drink containing more than 1% of alcohol. Drinking has been naturally occurring and has evolved into a variety of unique forms depending on region, ethnicity, climate, climate and cultural differences. Since there was no modern fermentation technology before the existence of bacteria, yeast, etc., The fermented bean was prepared by the native yeast present in the yeast. However, there is a problem in that it is difficult to maintain good and constant efficacy by using native yeasts. In order to solve this problem, yeast having excellent characteristics among the native yeasts in modern times was selected and separated to carry out fermentation, and excellent and constant efficacy was maintained to produce excellent fermented beverage.

Alcohol fermentation of sugar (starch) is essential for the production of alcohol, and yeast suitable for the characteristics of alcohol is used for this purpose. The beer has cohesion for low temperature fermentation and reuse, whiskey for high temperature fermentation and alcohol production, alcohol and alcohol for alcohol and spirits, and at the same time high alcohol production ability as fermentation characteristics. Whiskey, brandy, rum, gin, vodka, tequila and Japanese distilled liquor are all mainstream manufactured by carefully selected yeast, and research has been continuously carried out to find the best yeast for each mainstream characteristic and to industrialize it.

Distilled spirits are made by distillation to obtain higher alcohol content than fermented beverages. They include whiskey, brandy, rum, gin, vodka, tequila and distilled shochu, usually containing alcohol fractions of over 40 degrees Celsius. The distillation column among the distillation columns is obtained by mixing, fermentation and distillation of the raw material containing starch, starch and water. The raw materials of the distillery include cereals such as rice, barley and corn containing a lot of starch, and documents such as potatoes and sweet potatoes.

In this regard, Korean Patent Registration No. 10-0561563 relates to a method for producing distilled soju using a purified enzyme step by step, and does not use a germ in the liquefaction, saccharification and fermentation process of soju, A method for shortening the time required for shortening the time is short. In addition, Korean Patent Registration No. 10-0415307 has proposed a distillation column characterized by using an fermentation product made by inoculating and propagating white germ with a raw material in which peas are mixed in cereals.

However, the characterization of yeast for the production of distilled liquor in Korea has been limited. Yeast for the production of distilled liquor, which is currently used in industry, is mainly used for baking or yeast for wine, alcohol, whiskey and Japanese liquor. As the product is imported and used, the manufacturing cost is increased.

The object of the present invention is to develop distilled liquors having excellent sensory characteristics and to develop distilled liquor-specific yeasts. That is, an object of the present invention is to find a yeast suitable for producing a distilled liquor, and to produce a distilled liquor exhibiting excellent flavor and excellent sensory characteristics by using the yeast strain. It is also intended to replace industrialized imported yeast used for the production of distilled spirits.

Accordingly, the present inventors have conducted intensive studies to achieve the above object. As a result, they have found that the strain Saccharomyces cerevisiae 88-4 has excellent fermentation characteristics for producing distilled liquor, and the sensory properties And thus the present invention has been completed.

The distilled liquor prepared according to the present invention has excellent sensory characteristics due to proper harmony of flavor contributing ingredients by using the strain Saccharomyces cerevisiae 88-4 when fermenting alcohol.

1 is a flow chart of a method of manufacturing a distillation column according to an embodiment of the present invention.
Figure 2 compares base sequences CBS405 and 18S rRNA sequences of the same species as Saccharomyces cerevisiae 88-4 of the present invention.

Hereinafter, the present invention will be described in detail. However, it should be understood that the present invention may be embodied in many other specific forms without departing from the spirit or essential characteristics thereof.

In one aspect, the present invention provides novel Saccharomyces cerevisiae 88-4 88-4).

The inventors of the present invention evaluated yeast strains which were isolated from various yeast strains by collectively evaluating alcohol efficacy, occurrence of acid bubble formation, fragrance components and sensory test results, And it was confirmed that Servebiia. The fermentation strain 88-4 according to the present invention is a wild brewing yeast isolated from a traditional yeast, and has excellent fermentation characteristics for producing a distilled liquor. The results of this study are as follows: (1) It was confirmed that the isolates were different from the known strains of the same species and the industrialized strains. This strain was named as 88-4 strain and deposited on October 2, 2013 with the deposit number KCCM11456P at the Korean Society for Microbiological Research.

In another aspect, the present invention relates to a distillate produced using the Saccharomyces cerevisiae 88-4.

In another aspect, the present invention relates to a process for preparing a distillate using the Saccharomyces cerevisiae 88-4. The process for producing a distillate of the present invention comprises a) an inlet manufacturing step, b) a first fermentation step, c) a second fermentation step, d) a distillation step and e) a filtration step.

In one embodiment, step a) comprises admixing and cultivating white germ with the grains grown to produce entrances. In step (b), the yeast culture solution and water are mixed with the entry prepared in step (a) and fermented to prepare a first fermentation broth. In step c), the fermented starch-containing raw material and water are mixed with the primary fermentation broth prepared in b) and fermented to prepare a secondary fermentation broth. In step d), the secondary fermentation broth prepared in step c) is distilled.

In one embodiment, Saccharomyces cerevisiae 88-4 is used for fermentation in step b), step c) or steps b) and c). Preferably, Saccharomyces cerevisiae 88-4 is used for both fermentation in steps b) and c).

A flow chart of a method of manufacturing a distillation column according to an embodiment of the present invention is shown in FIG. Referring to FIG. 1, an embodiment of a method of manufacturing a distillate of the present invention will be described step by step.

In order to prepare a distilled liquor to which Saccharomyces cerevisiae 88-4 strain according to the present invention is applied, entry is first made. Entry can be made by conventional methods by inoculating white rice bran into cereals such as rice or barley. For example, inoculation can be made by inoculating white rice germ in a rice grate and propagating white rice germs in a thermo-hygrostat for 48 hours while maintaining the temperature at 33 ° C and a humidity of 95%.

An aspherical microorganism, preferably Aspergillus luchuensis , is used as the white germ, but is not limited thereto.

The novel yeast strain according to the present invention can be cultured in a large amount by a conventional method for culturing Saccharomyces sp. As the culture medium, a medium composed of carbon source, nitrogen source, vitamins and minerals can be used. For example, a yeast malt extract agar medium, a potato dextrose agar (PDA) medium, or an immobilized glycoside can be used as a medium. The immigration saccharified liquid is added to 300 parts by weight of water with respect to 100 parts by weight of the immigration using the immigration produced in the above step, stirred and saccharified at 60 to 65 ° C for 3 to 5 hours and then at 4,500 to 6,000 rpm for 20 to Centrifugation for 30 minutes, filtration, sterilization, and dilution to 10 ° Brix to 14 ° Brix. The culture can be carried out using the culture medium under aerobic conditions at a temperature of 25 ° C to 30 ° C for 36 hours to 72 hours and then stored at 4 ° C.

Water is added to the impregnation prepared in the above step and mixed, and the temperature is adjusted to 25 ° C. It is preferable that 120 to 140 parts by weight of water are added to 100 parts by weight of the above-mentioned mixture. When the number of cultured yeast cells is 10 ⁸ / ml or more per 100 parts by weight of the entry, the primary fermentation can be carried out at 25 ° C for 4 days to 6 days after inoculating 1.5% to 2.5%

The primary fermentation broth prepared in the above step and the fermented starch-containing raw materials are mixed and fermented to prepare a soup base.

The starch-containing raw material may be at least one selected from the group consisting of rice, barley, corn, millet, wheat, potato and sweet potato, but is not limited thereto. It is preferable that the fermented starch-containing main raw material and water are mixed with the primary fermentation broth prepared in the above step and then the secondary fermentation is carried out at a temperature of 25 ° C to 30 ° C for 8 to 12 days.

The secondary fermentation broth obtained in the above step is heated to produce a steam, and the produced steam is cooled to produce a distillate. The distillation may be a normal pressure distillation or a reduced pressure distillation using a commonly used single distillation column. The atmospheric distillation can be distilled at 80 to 120 DEG C to 760 mmHg (atmospheric pressure), the intermediate pressure distillation can be distilled at 360 to 400 mmHg at 80 to 100 DEG C, and the vacuum distillation is performed at 70 to 150 mmHg It can be distilled.

Thus, in another aspect, the present invention provides a distillate produced according to the above process.

The distilled liquor prepared according to the present invention has excellent sensory characteristics due to proper harmony of flavor contributing ingredients by using the strain Saccharomyces cerevisiae 88-4 when fermenting alcohol.

The present invention will be described in more detail by way of the following examples. However, the following examples are for the purpose of illustrating the present invention and are not intended to limit the scope of the present invention.

[Example]

1st round selection

The first selection criteria were alcohol efficacy and no bubble formation. Among the yeast - preserved yeast isolated from the nationwide collected nuruk, white rice germ (Aspergillus lucensis) was inoculated into the rice grown and maintained at 33 ° C and 95% humidity in a thermo - hygrostat for 48 hours. Were cultured in 100 ml of the immobilized glycosaminoglycan at a concentration of 2 × 10 ⁷ / ml of yeast, and 108 yeast strains which had alcohol efficacy and did not produce lipid membrane-derived bubbles during fermentation were first selected.

Second selection

The second selection was selected according to the excellent yeast selection criteria shown in Table 1 below. A total of 108 strains were selected and fermented at 25 ℃ for 5 days. The alcohol content was more than 12% (v / v) and the pH was 3.5 (w / v) 40 strains were selected under the conditions of ± 0.2, an acidity of 15.0 ± 3.5 ml (0.1 N NaOH titration), a sugar content of 10 ± 3 Brix (%) and a viable cell count of primary fermentation yeast of 2 × 10 ⁷ / ml or more.

3rd round selection

The third selection was selected according to the excellent yeast selection criteria shown in Table 2 below.

Forty strains of the second selection were immersed in a volume of 2,000 ㎖ to confirm fermentation characteristics for the production of distilled liquor. The number of viable cell counts and alcohol production of primary fermentation yeast, primary alcohol production of secondary fermentation, and final alcohol content of fermentation end were analyzed. Alcohol fermentation patterns were analyzed for the selection of fast - forming yeast for the production of distilled liquor. The fast dominance of yeast in the fermentation stage for distillers is a very important factor in the efficient use of the substrate and in the control of flavor contributing ingredients. Alcoholic fermentation patterns were analyzed by increasing the number of differentiated alcohol and divided into two groups: first half fermentation group and second half fermentation group. Among the yeasts showing a fermentation pattern in which the first half increases rapidly and the second half becomes gentle by drawing a polynomial function graph of the differentiation alcohol production, the second fermentation third day point alcohol is more than 8% (v / v) and the fermentation alcohol is 15% v / v). As described above, 6 strains were selected by comparing alcohol content, pH, acidity, amino acid degree, alcohol fermentation pattern analysis, final alcohol content in secondary fermentation end, sugar content in supernatant Brix, and unfermented free sugar equivalent (g / (Table 3).

Final selection

The final selection was made by analyzing the fragrance components of the distillate and evaluating the preference of the distillate for 6 strains and 3 industrialized strains. Among the industrialized yeasts, three types of industrialization strains were used: Fermentis Ethanol Red Yeast, Whimsy (Fermentis M1 Yeast), and Yeast for Manufacturing in Japan (Brewing Association Suzhou No.2).

Manufacture of distilled spirits

The distillate was prepared for 9 strains. For the manufacture of the immigration, the rice was immersed for 3 hours, drained and then added. In 10 kg of the expanded rice, 2 g of the spore forming material (Aspergillus lucensis) spore was inoculated and cultured for 48 hours at 33 캜 and 95% humidity.

For the first fermentation, 513 g of the above-prepared entrant and 718 g of water were mixed, followed by culturing in an adherent glycoside to inoculate 10 ml of yeast which was confirmed to have a water content of 10 ⁸ / ml or more. As the yeast culture, 200 g of the immigration produced by the above method and 600 g of water were mixed and stirred at 65 캜 for 3 hours and 30 minutes. After saccharification, the mixture was centrifuged at 4,700 rpm for 30 minutes, filtered through filter paper, and diluted with water to 10 ° Brix. The above mixed solution was fermented at 25 DEG C for 5 days to prepare a primary fermentation broth.

Rice was prepared for secondary fermentation. In order to remove foreign matter or contamination of rice, it was cleaned with water and immersed in water at room temperature for 3 hours for sufficient absorption of water, and then steamed for 60 minutes at normal pressure. After the addition, the mixture was cooled at room temperature for 20 minutes. 1,743 g of water and 1,026 g of rice were added to the primary fermentation broth prepared above, and the mixture was fermented at 25 캜 for 10 days to prepare 4,000 g rice syrup.

The rice syrup was distilled under reduced pressure at 110 mmHg at an internal temperature of 40 ° C using a single distiller designed as a supercritical fluid. The decompression pump was a water pump. The distiller for the distillation experiment consisted of a round shaped flask with 5 ℓ capacity of frit, a frost prevention distillation column, a horizontal cooler tube, a sight glass with graduated pressure reducing valve, and a receiver to collect the effluent. The initial fermentation volume for distillation was 2 L, the initial fermentation broth temperature was 20 to 25 DEG C, and the cooler for cooling the effluent vapor was set to 5 DEG C and circulated at a rate of 15 L / min. During the distillation, the temperature of the effluent that passed through the cooler was adjusted normally to 20 to 25 ° C and the maximum effluent temperature to not exceed 30 ° C. Distillation rate and effluent temperature were controlled by heat input to the still. The spirits were kept in the mainstream without breaking the flow of freshwater, so that the alcohol content was about 45%. As a result, a distillation stock solution with an alcohol content of 45% was obtained.

Fragrance component analysis

The fragrance components were analyzed by GC-MS. Since the fragrance component is expressed by the peak area ratio, the distillate alcohol was batch diluted with 20% v / v and the analysis was carried out with the same alcohol concentration. 20 ml of the sample was equilibrated at 60 ° C, perfumed by SPME (solid phase microextraction) method and collected by Hewlett-Packard equipped with a Stabilwax ^® -DA column (30 m × 0.25 mm Id × 0.25 μm film thickness, manufactured by Restek Corp.) Packard 7890A GC / HP-5973N mass selective detector (MSD) (manufactured by Hewlett-Packard Co., USA). The oven temperature of the GC was increased from 50 ° C to 200 ° C, the injector temperature was 250 ° C, and helium was used as the carrier gas. The MSD conditions include capillary direct interface temperature of 250 ° C, ion source temperature of 230 ° C, ionization voltage of 70 eV, mass range of 33-350 amu And the scan rate was 2.2 scan / sec. Identification of volatile flavor components was confirmed by comparing retention indices (RI), mass spectra and aroma properties. The analyzed results (unit peak area%) are shown in Table 4 below.

Sensory evaluation

The degree of preference of the distillate thus prepared was evaluated. For the sensory evaluation, the distillate was prepared with an alcohol content of 20% (v / v). (9 points: very good; 7 points: good; 5 points: average; 3 points: not good; 1 point: very poor). The evaluators were repeatedly evaluated by 9 lecturers on two occasions, and a brief description of the evaluation was given. The results are shown in Table 5 below.

As a result, the preference of spirits made from 88-4 was the highest and the preference was higher than industrialized yeast at 95% confidence level. As a result of the comparison of yeast characteristics and sensory evaluation for the production of distilled liquor, 88-4 strain was finally selected.

Identification of the strain

The final selected 88-4 strain was analyzed for 18S rRNA sequence (using ITS1 and ITS4 primers) and showed 99% homology with the genbank accession number EF620858.1. It was identified as Myes serebisji. Fig. 2 shows the results of comparing 18S rRNA sequences between CBS405 and 88-4 strains of the strain Saccharomyces cerevisiae. In addition, the 18S rRNA sequence of strain 88-4 is shown in SEQ ID NO: 1.

Korea Microorganism Conservation Center KCCM11456P 20131002

<110> KOREA FOOD RESEARCH INSTITUTE <120> FLAVOR PROMOTED DISTILLED SPIRIT USING YEAST SACCHAROMYCES          CEREVISIAE 88-4 <130> IPDB-50915 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1167 <212> DNA &Lt; 213 > 18S rRNA of Saccharomyces cerevisiae 88-4 (KCCM11456P) <400> 1 taagtgcgac agatcggtca ggttaactaa tgcagtacgg atcggtggca atatgatagt 60 aatgtggtgt gaacctgccg atgtcgaagc aaggaaaggt caagaatgaa ctccaggtgt 120 gtgtgtaccg tttaatgtgc cataacaaaa gacacaatct ctcgccgctg ggtagcacga 180 aagagaacta cagactagca agacgcgcac ctaagcgcag gcgcggctcg tactctctca 240 tctcttgtct tctcgcgcag tgtaagcgtc tctatgctct cgccaaaaaa aaaaaaaaaa 300 aatccacttt caaaaaaaat taaaaatctc ttaaaaggcc cctccggggg gggggggctc 360 cggaggatca ttaaagaaat tttataattt tggaaaaggg tttttttttt tttgttttgg 420 caagagcatg agagctttta ctgggcaaga agacaagaga tggagagtcc agccgggcct 480 gcgcttaagt gcgcggtctt gctaggcttg taagtttctt tcttgctatt ccaaacggtg 540 agagatttct gtgcttttgt tataggacaa ttaaaaccgt ttcaatacaa cacactgtgg 600 agttttcata tctttgcaac tttttctttg ggcattcgag caatcggggc ccagaggtta 660 acaaacacaa acaattttat ctattcatta aatttttgtc aaaaacaaga attttcgtaa 720 ctggaaattt taaaatatta aaaactttca acaacggatc tcttggttct cgcatcgatg 780 aagaacgcag cgaaatgcga tacgtaatgt gaattgcaga attccgtgaa tcatcgaatc 840 tttgaacgca cattgcgccc cttggtattc cagggggcat gcctgtttga gcgtcatttc 900 cttttcaaac attctgtttg gtagtgagtg atactctttg gagttaactt gaaattgctg 960 gccttttcat tggatgtttt ttttttccaa agagaggttt ctctgcgtgc ttgaggtata 1020 atgcaagtac ggtcgtttta ggttttacca actgcggcta atctttttta tactgagcgt 1080 attggaacgt tatcgataag aagagagcgt ctaggcgaac aatgttctta aagtttgacc 1140 tcaaatcatg tagaggccca gtccttg 1167

Claims (6)

A method for producing Saccharomyces cerevisiae 88-4 ( Saccharomyces cerevisiae) 88-4, Accession No. KCCM11456P):
a) mixing and culturing white germ with the added cereal to produce an entry;
b) mixing the fermentation broth with water to prepare a first fermentation broth by fermentation;
c) mixing a primary fermented broth with starch-containing raw material and water to produce a secondary fermentation broth;
d) distillation stage; And
e) Filtration step. The method according to claim 1,
Wherein the starch-containing main raw material is at least one selected from the group consisting of rice, barley, corn, millet, wheat, potato and sweet potato. The method according to claim 1,
Wherein the saccharomyces cerevisiae 88-4 is used for the fermentation of step b) and step c). A distillate produced according to the process of any one of claims 1 to 3. Saccharomyces sp. 88-4 ( Saccharomyces cerevisiae 88-4, Accession No. KCCM11456P). Saccharomyces sp. 88-4 ( Saccharomyces cerevisiae 88-4, accession no. KCCM 11456P).

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