KR101284263B1 - Saccharomyces cerevisiae NK28 for producing wine and method for producing wine using the same - Google Patents

Abstract

The present invention relates to a new yeast strain Saccharomyces cerevisiae NK28 (NK28) (accession number KACC 93117P), which is isolated from kiwi shells, for wine casting. Saccharomyces cerevisiae NK28 strain of the present invention has excellent utility for disaccharides, especially maltose, compared to the commercially available yeast strains for wine fermentation, and can be grown in a medium containing 30% glucose as well It produces a large amount of alcohol in a short time, so it has excellent alcohol efficiency. Therefore, Saccharomyces cerevisiae NK28 strain of the present invention has a high sweetness and alcohol content, but the taste is soft, it can be usefully used to prepare wines to suit the taste of Korean people.

Description

Saccharomyces cerevisiae NK28 for producing wine and method for producing wine using the same}

The present invention relates to a novel strain Saccharomyces cerevisiae NK28 (Skcharomyces cerevisiae NK28) (Accession No. KACC 93117P) and a wine manufacturing method using the same. More specifically, the present invention relates to a wine-making Saccharomyces cerevisiae NK28 strain isolated from kiwi skin, and a method of producing wine having high sugar and alcohol content and soft taste using the same.

Recently, with increasing income and interest in well-being in Korea, the consumption of wine is growing very fast by 20-30% every year. However, most of the wine still depends on imports, the imports are constantly increasing. Specifically, Korea's wine imports grew 128.6% between 2002 and 2006. In 2011, a total of 37.6 million bottles were imported, accounting for 81.5% of the total wine consumption. Accordingly, many studies on wine production are being actively conducted in Korea, and production of wines that suits Korean tastes is being attempted with Campbell Early and Geobong grapes, which are familiar to Korean consumers.

However, since the quality of wine produced in Korea is inferior to that of imported wine, there is little competitiveness, and the demand for domestic wine is insignificant compared to imported wine, and there are many parts that need to be improved in quality. Domestic wines are mainly produced by blending imported wines. The quality of wine is influenced by many grape varieties, but the fermentation strains, the manufacturing process and the ripening process are also affected. Korea is not environmentally qualified to grow high quality grapes compared to France and Chile, and since it takes a considerable time for varieties to be improved, the development of excellent grape varieties has a strategy to be competitive in the short term. I can't. Therefore, in order to improve the quality of wine using domestically grown grape varieties, many studies on fermentation yeast and manufacturing process are required. In particular, in the production of wine, the fermentation capacity and the flavor component of fermentation are mostly determined by fermented yeast as a factor affecting the quality. Nevertheless, since the strains used in the domestic wine production were mostly fermented yeasts such as S. cerevisiae Lv001 and S. cerevisiae La Parisienne, the need for developing fermented yeasts suitable for domestically grown grape varieties is increasing.

When using domestically produced grapes for wine production, the sugar content is low (usually 13-14 brix), high acidity, high content of malic acid has a strong and acidic disadvantages. Koreans tend to prefer sweetness to bitterness when drinking wine. However, when fermentation of wine by adding excess sugar at one time, most of the fermentation strains are killed by osmotic shock and thus the quality of the wine is degraded. Therefore, a fermentation yeast capable of withstanding osmotic pressure due to high sugar concentration is required. In addition, alcohol is produced when the yeast strain fermented with glucose in grape juice. At this time, if the fermented yeast ferments sugar at a very high rate and produces alcohol rapidly, the yeast strain is killed before the fermentation is sufficiently completed, thereby affecting the quality of wine and alcohol content. If fermented yeast efficiently utilizes maltose, which has two glucoses as its carbon source, it takes longer to make maltose and use it, thus relieving the impact of high glucose content and the impact of rapid alcohol production for longer. Fermentation can be induced.

Therefore, the present inventors have been separating the wild type yeast strains from various fruit peels in order to solve the problems of domestic wines as described above and to prepare a fermented yeast suitable for the production of high sugar sweet wines according to the taste of Korean people. The present invention was completed by identifying Saccharomyces cerevisiae NK28.

It is therefore an object of the present invention to provide a novel yeast strain Saccharomyces cerevisiae NK28 which can produce a large amount of alcohol in a faster time than conventional yeast strains.

In addition, another object of the present invention to provide a method for producing wine using the yeast strain.

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

In order to achieve the object of the present invention as described above, the present invention is a wine casting new strain Saccharomyces cerevisiae NK28 ( Saccharomyces cerevisiae NK28) isolated from the kiwi skin (Accession number KACC 93117P; Agricultural genetic resource centers).

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In one embodiment of the present invention, the strain may use maltose as a carbon source.

In one embodiment of the present invention, the strain may be a wine fermentation strain that does not occur aggregation during the culture.

In another aspect, the present invention provides a method for producing wine, characterized in that using the new strain Saccharomyces cerevisiae NK28 for wine casting separated from the kiwi skin.

In one embodiment of the present invention, the wine production method may be fermented grapes with starch of cereals.

In one embodiment of the present invention, the grains may be rice.

The present invention further provides a wine produced by the method of the present invention.

Saccharomyces cerevisiae NK28 strain of the present invention has excellent utility for disaccharides, especially maltose, compared to the commercially available yeast strains for wine fermentation, and can be grown in a medium containing 30% glucose. In addition, it produces a large amount of alcohol in a short time, and excellent alcohol efficiency. Therefore, Saccharomyces cerevisiae NK28 strain of the present invention has a high sweetness and alcohol content, but the taste is soft, it can be usefully used to prepare wines to suit the taste of Korean people.

1 is a phylogenetic tree isolated from Saccharomyces cerevisiae NK28 strain and fruit of the present invention, based on the association of 26S rDNA sequences of other yeast strains with high glucose tolerance, alcohol resistance, and maltose availability.
Figure 2 is an optical micrograph comparing the cell morphology of wine fermented yeast commercially available with Saccharomyces cerevisiae NK28 strain of the present invention (magnification 400X). (A) Saccharomyces cerevisiae NK28, (B) Saccharomyces cerevisiae Lv001, (C) Saccharomyces cerevisiae La Parisienne.
Figure 3 is a graph comparing the glucose tolerance and maltose availability of the Saccharomyces cerevisiae NK28 strain of the present invention with that of the commercial wine fermentation yeast.
4 is a graph comparing the alcohol resistance of Saccharomyces cerevisiae NK28 strain of the present invention with that of the commercial wine fermentation yeast.
5 is a graph comparing the alcohol production capacity of Saccharomyces cerevisiae NK28 strain of the present invention with that of the commercial wine fermentation yeast.

The present invention is characterized by providing a novel strain Saccharomyces cerevisiae NK28 (Accession No. KACC 93117P), which is a novel strain suitable for preparing a soft sugar of high sugar content in Korea.

On the other hand, fermented yeasts used for alcohol fermentation are known to have good fermentation activity at a temperature of about 30 ° C. and to produce approximately 12% of alcohol, although there are some differences depending on the substrate. (Chul S Shin, Analysis of The Factors which Enhance Yeast Growth and Ethanol Production, The State University of New Jersey, USA, P5, 1984).

In addition, the growth of yeast during alcohol fermentation is controlled metabolism by the content of the carbon source and nitrogen source in the substrate, and thus the population of the yeast proliferates by proliferation such as germination. However, will receive the inhibition of growth depending on the metabolite, alcohol content, in the above a certain concentration of alcohol the growth is considerably inhibited. (A. Lentini, The Impact of Ethanol Stress on Yeast Physiology, Brewing Yeast Fermentation Performance 2 ^nd Edition, UK , P25, 2003)

Therefore, when the fermentation strain during the fermentation process using a strain having a relatively low growth inhibition for alcohol, a metabolite by the yeast can obtain a high fermentation yield by the same raw material to increase the productivity.

In addition, when fermented wine, especially when the wine is fermented by adding an excess of sugar at the same time, most of the fermentation strains are killed by osmotic shock and thus the quality of the wine. Therefore, the production of high sugar sweet wine requires fermented yeast capable of withstanding osmotic pressure due to high sugar concentration.

Therefore, the present inventors isolated Saccharomyces cerevisiae NK28 ( Saccharomyces cerevisiae NK28) as a result of searching for a new yeast strain that satisfies these conditions, and deposited on February 07, 2011 to the National Institute of Agricultural Sciences Microbial deposits have been granted (Accession No. KACC 93117P).

In more detail, the present inventors separated the yeast attached to the surface of various fruits, such as grapes, strawberries, kiwi, etc. purchased in Korea in order to separate the new strains, and secured 637 wild yeast strains. Saccharomyces, which can be widely commercialized because they have characteristics that are commonly desired as wine yeast as a result of analyzing 120 characteristics of alcohol resistance, maltose availability, alcohol production ability, and then using genetic information. Saccharomyces cerevisiae strains were identified and named Saccharomyces cerevisiae NK28 (see FIG. 1).

The inventors of the Saccharomyces cerevisiae NK28 ( Saccharomyces cerevisiae NK28) Saccharomyces cerevisiae , which is commercially available as wine fermentation yeast, to find out whether the strain has superior properties compared to conventional wine fermentation yeasts. Lv001 ( Saccharomyces cerevisiae Lv001) and Saccharomyces cerevisiae, which was developed as a fermented yeast for bread but is also used for wine fermentation as it is effective in reducing the sourness of wine during wine making. La Parisienne ( Saccharomyces cerevisiae And comparative experiments with La Parisienne).

First, the morphological characteristics were compared with those of the above strains. In the NK28 strain, the cells grew into individual single cells, but the La Parisienne strain showed that the cells aggregated and grew together (FIG. 2). Therefore, it was found that NK28 is not the same strain as La Parisienne strain, and when fermented with NK28 strain without aggregation, the sedimentation rate is slower, so it is more efficient for wine fermentation than wine fermentation with Parisienne strain with cell aggregation. I could see that.

In addition, in terms of sugar utilization, NK28 is glucose, galactose, methyl glucopyranoside (methyl-aD-glucopyranoside), maltose, sugar (D-saccharose, sucrose), trehalose (D-trehalose), melezitose (D-melezitose) and Raffinose (D-raffinose) can be used as carbon sources, whereas Lv001 uses glycerol, which is rarely used by Saccharomyces cerevisiae strains, and methylglucopyranoside ( methyl-aD-glucopyranoside) was not used (Table 1).

Therefore, NK28 new strain of the present invention is characterized by the efficient use of maltose as a carbon source, it takes a long time to make maltose into glucose and then use it, the impact of high glucose content and the impact of rapid alcohol production To alleviate grape juice fermentation for longer.

In addition, according to another embodiment of the present invention, the degree of glucose tolerance for the new strain of the present invention was analyzed, as a result, growth of NK28 strain and other strains in YPD liquid medium containing 40% glucose Although there was no significant difference, the NK28 strain showed higher growth than Lv001 and La Parisienne strains when 30% of glucose was contained (FIG. 3).

In addition, the alcohol fermentation test results, the NK28 strain showed an increased alcohol production than the Lv001 strain, it can be seen that produced more alcohol of 25% at 72 hours compared to the La Parisienne strain (Fig. 5).

Therefore, Saccharomyces cerevisiae NK28 ( Saccharomyces of the present invention) cerevisiae NK28) (Accession No. KACC 93117P) strain can be used for the production of wines with high sugar content and high alcohol content and soft taste.

Furthermore, the present invention can provide a method for producing wine using Saccharomyces cerevisiae NK28 strain of the present invention.

Production of wine using Saccharomyces cerevisiae NK28 of the present invention can be carried out according to the production method of fermented strain using a known yeast strain. After harvesting the grapes, the grapes are crushed, pressed and filtered to obtain fruit juice, and then sugar or glucose is added to adjust the sugar to a suitable sugar.Also fermented with alcohol, yeast is added to make fermented liquor. It is common to produce it by commercialization.

In one embodiment of the present invention, the production of wine using the strain according to the present invention, using a strain having excellent properties such as the maltose availability and sugar resistance, such as the addition or addition of additives is common in the grape juice medium It can be made by fermentation method. The additive may include starch of grains including rice, water, a small amount of saccharase, sulfurous acid, cloves (丁香, clove) and the like, and a small amount of lactic acid may be added to prevent contamination. Further, after the incubation, the wine can be obtained by further purifying the fermentation broth obtained by fermentation at a constant temperature. When rice is added to the wine, high alcoholic wine can be produced, and this wine can be drunk without objection to the taste of Koreans tamed by the taste of Takju. In the case of general wine fermentation, when a suitable amount of sulfurous acid is added, wine fermentation bacteria are added, and when fermentation starts, the existing wine fermentation bacteria disappear and the added wine fermentation bacteria grow. Fermentation corresponding to the oxidation reaction generates heat, so it must be adjusted to a certain temperature. On the other hand, it is necessary to prevent the inflow of oxygen to prevent acetobacter, which is an acetic acid bacterium that makes alcohol into vinegar, so that the yeast fermentation can proceed smoothly. When the fermentation proceeds sufficiently and the alcohol concentration reaches 7-15%, a compound such as bentonite is added to filter out the suspended matter. This is called a racking process. The wine is then placed in a glass bottle, covered with a cork, and aged in a dark, moderately controlled warehouse. Taste and aroma are added by the complex reaction of the compounds remaining in the wine during the aging process. On the other hand, Korean Patent Application No. 2005-0107598 ("Sugar prepared by adding yeast and cloves without using yeast and its preparation method", filed Nov. 10, 2005) is mixed with yeast by adding clove and fermented together. It inhibits fermentation and at the same time gives a soft taste to alcohol and promotes alcohol fermentation to increase its concentration by 2 ~ 4%, and by adding clove with yeast in wine making, it makes the wine soft and sour taste and softens the taste. It is described that the concentration can also be increased. Specifically, the manufacturing method is a hydration process of rice; Malt preparation process; Steaming process of hydration rice; Mixing process of malt water and soybean rice; Sweetening and saccharification processes; Saccharification filtrate manufacturing process; Yeast and cloves addition process; And alcohol fermentation and cold aging process.

Meanwhile, Korean Patent Application No. 2004-0056243 ("Wine Manufacturing Method Using Starch Sugar", filed Jul. 20, 2004) uses starch raw material such as starch or rice to prepare high sugar content starch sugar by high temperature liquid saccharification method. A method of making wine using this starch sugar solution is described. Here, the high temperature liquid saccharification method is a method of making pure saccharification liquid, and when liquefied enzyme and protease are added to starch raw material and heated to high temperature, various harmful bacteria and wild yeast are almost killed. Refers to a method of preparing starch sugar solution in a relatively quick time. Specifically, the method for preparing starch sugar solution by high temperature liquefaction saccharification method is as follows. First, starch raw materials such as rice and rice are dissolved in water and put into a liquefaction tank. The pH is obtained by adding co-enzyme of Aspergillus sp. And Bacillus sp. Adjust 6.0 to 6.5 to stabilize the liquefied enzyme. Although not limited to this, aspergillus coenzyme includes Aspergillus Usamii, Aspergillus Niger, Aspergillus Kawachi, and the like. The next step is to raise the temperature of the liquefaction tank to 96 ~ 98 ℃ and maintain it for 1 hour.When the temperature is lowered to 60 ℃, aspergillus coenzyme, which is a coenzyme that produces a short enzyme and a saccharifying enzyme, is produced. Add the enzyme and Lycopus coenzyme at the same time and keep for 6-8 hours. Thereafter, when the temperature is rapidly lowered to reach 30 ° C., a starch sugar of 25 ~ 35 Brix can be obtained. Inoculate yeast here and hold for 1 to 2 days at 23 ~ 25 ℃, then add grape crushing solution containing potassium metasulfite and pectinase in meta and ferment at 23 ~ 25 ℃ for 5 ~ 6 days. . After pressing the fermentation broth and filtered through diatomaceous earth filtration or filter press, and put into a storage tank and then fermented after 2 ~ 4 days at 13 ~ 15 ℃, 4 ~ 6 days at 6 ~ 8 ℃. After completion of the post-fermentation, tartaric acid is separated by refrigeration at 0-4 ° C. for 4-7 days, and sedimentation is performed twice a week to obtain the desired wine.

In addition, Korean Patent Application No. 2003-0083245 ("Starch Wine and Manufacturing Method", filed Apr. 19, 2002) discloses a pulp in a fructose fermented with raw material which is primarily fermented with no steamed starch raw material, crude glycosylase, and cultured yeast. ) And a method of preparing starch wine by filtration of the grape juice without filtering the fruit and secondary fermentation. As the starch raw material may be used uncooked raw rice or raw corn moisture, or may be used by mixing raw rice and raw corn starch. By omitting the steaming step, the decomposition or destruction of nutrients in the starch raw material by heat can be prevented and the deterioration of the taste can be prevented. Grapes are ground and crushed and then crushed and used without pulp and filtration. The pulverized grape juice may be partially degraded by the action of pectinase in the fruit juice, and at this time, pectinase may be artificially added to promote degradation of the skin and the flesh. In general, grain wine is inoculated with molds (aspergillus, etc.) that produce saccharifying enzymes in the gourd rice, which is made by increasing the amount of grains, and then, when maintained at an appropriate temperature, starch turns into sugar and secondly, sugar turns into alcohol. The wine is made through fermentation by yeast growing on grape skin after removing fruit and seeds, but the starch wine according to the present invention is firstly fermented as a sugar-free starch raw material. When enough sugar and alcohol are produced so that various bacteria that grow on the grapes themselves cannot grow, they are prepared through the second fermentation process by adding crushed grape juice containing the flesh and skin as it is. Starch wine according to the invention is characterized in that it is fermented raw (raw) without increasing the starch raw material. By omitting the capital increase process, not only the possibility of contamination of various germs that may occur in the cooling down stage can be excluded, and production costs such as facility costs, fuel costs, and labor costs can be saved. Starch wine according to the invention is characterized in that, unlike the general wine fermentation of the juice after the juice, the juice and the filtration process is omitted and the juice of the flesh and the skin is crushed as it is, thereby adding the starch raw material In spite of this, the color and flavor of the wine can be maintained.

Meanwhile, Korean Patent Application No. 2001-0072830 ("Method of manufacturing distilled wine using grape grapes", filed Nov. 21, 2001) is prepared by distilling the fermented and aged grape stock solution, and then distilling and freezing the distillate again. One distilled wine and a method of making the same are described. Sugar or sugar is added to a general distilled wine production method, ie, grape juices, such as Delaware or Campbell, until the sugar content is reached to the calcined sugar, and then added to the fermenter, and then stored in a fermenter. The vinegar is fermented and matured for a certain period of time, at least 3 months to 1 year or more at room temperature, and the fermented and matured vinegar is distilled through a distillation process, and then pigments and spices are added again. The wine produced by the process of making a low alcohol distilled wine with dilution and cooling to lower the alcohol content to 6-12% is monotonous with two colors of white or red wine, and over time There is a problem that the original fragrance and taste that it had originally fall. Therefore, the method adds a certain amount of vinegar to the distilled wine after fermentation and maturation, and re-ages and refrigerates the wine, thereby maintaining a wine's original aroma and taste as well as improving the production of wine. to provide.

In addition to the above-described methods, all known processes for making wine or fermented wine can be used in the method for producing wine according to the present invention.

Furthermore, the present invention is characterized by providing a wine produced by the wine production method of the present invention.

As described above, the NK28 strain, a new strain according to the present invention, has high sugar and alcohol content and tastes higher than that of the wines sold because it does not have glucose tolerance, has excellent alcohol producing ability, and does not cause aggregation of cells during fermentation. This is a soft feature.

Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited to these examples.

< Example  1>

Sakaromayses Selivijia NK28  Isolation and Selection of Strains

The skins of unwashed fruits purchased in Korea were YPD broth [YPD broth; Add 1% yeast extract, 2% peptone, 2% glucose, mix well, and spread on YPD solid medium [1.5% agar added to YPD liquid medium] and incubate at 30 ° C for 72 hours. It was. 637 isolates identified as yeast by screening strains after screening with colony-like colony-like colony-type colonies on a new YPD solid medium two or more times. Was incubated again and stored at -80 ° C.

The isolated yeast strains were each tested for appropriate glucose tolerance, alcohol resistance, maltose availability, and alcohol production capacity as wine fermentation bacteria, respectively.

< Example  2>

Sakaromayses Selivijia NK28  Identification of the strain

In order to identify the selected strains, the chromosomal DNA of yeast isolates was extracted and the 26S rDNA gene region was amplified by PCR to determine the sequence and NCBI database (National Biotechnology Information Center http: //www.ncbi.nlm.nih). .gov ) to identify blasts using a blast search.

The primers used to amplify the 26S rDNA gene were NL-1 (5'-GCATATCAATAAGCGGAGGAAAAG-3 '; SEQ ID NO: 1) and NL-4 (5'-GGTCCGTGTTTCAAGACGG-3'; SEQ ID NO: 2), and PCR was performed at 94 ° C. After pretreatment for 3 minutes, 1 minute (denaturation) at 94 ° C, 1 minute (recombination) at 45 ° C, 2 minutes 30 seconds (extension) at 72 ° C were repeated 30 times, and final expansion was performed at 72 ° C for 10 minutes. . As a result, amplified 560 bp DNA fragment of 26S rDNA gene was confirmed by agarose gel (1%) electrophoresis and purified by PCR purification kit (SolGent, Korea).

By comparison, using the Blast search program for: (//www.ncbi.nlm nih gov US National Center for Biotechnology Information http..) Determined the DNA sequence of the purified DNA and then (SolGent Inc., Korea), its homology NCBI I identified it. As a result, one of the isolates from the kiwi was Saccharomyces cerevisiae ). The strain was Saccharomyces ( Saccharomyces) cerevisiae ) was named NK28, and the strain was deposited with the Agricultural Genetic Resource Center of the National Academy of Agricultural Science on February 7, 2011 and was assigned accession number KACC93117P.

Saccharomyces cerevisiae NK28 showed excellent characteristics in glucose tolerance, alcohol tolerance, maltose availability, alcohol production ability, and the PCR sequence amplified 26S rDNA gene sequence of Saccharomyces cerevisiae strain The phylogenetic association with other strains that were 100% identical to the gene of interest and selected and identified is shown in FIG. 1.

In order to compare the properties of Saccharomyces cerevisiae NK28 strain of the present invention as a wine fermentation strain, it is widely used as Saccharomyces cerevisiae Lv001 and bread fermentation strains that are already commercially available as wine fermentation strains. Saccharomyces cerevisiae La Parisienne, which is also used for wine fermentation, was selected and compared in the following examples.

< Example  3>

Sakaromayses Selivijia NK28  Morphological characteristics of the strain

Morphological characteristics between Saccharomyces cerevisiae L Paris001 and Saccharomyces cerevisiae Lv001 and NK28 strain of the present invention, which are currently used as wine fermentation yeast, were observed by optical microscopy while inoculating them in YPD liquid medium. Compared.

Figure 2 is respectively Saccharomyces ( Saccharomyces) cerevisiae ) NK28, Lv001, LaParisienne strains were observed by light microscopy (X400). As shown in the photo, NK28 and Lv001 strains were observed to grow cells as individual single cells, but La Parisienne strains were repeatedly observed to aggregate cells. Thus, it can be seen that NK28 and La Parisienne are not the same strain.

In general, it is known that fermentation of wine with La Parisienne strain yields a softer taste of wine than fermentation with Lv001 strain, and thus, it is expected that even when using NK28 strain, softer wine can be produced than when using Lv001 strain. Also, fermentation with NK28 strain that does not occur will be more efficient in fermenting wine than fermentation of wine with Parisienne strain, which causes cell aggregation because of the slow settling rate.

< Example  4>

Sakaromayses Selivijia NK28  Sugar in strain Usability  analysis

The sugar utilization characteristics of Saccharomyces cerevisiae NK28 were analyzed in more detail with the API 20C AUX kit (Biomerieux, France). The kit allows the yeast strain to test and identify biochemical abilities using 19 sugars. First, each strain was inoculated into YPD solid medium and incubated for 48 hours at 30 ° C., and then colonized with each strain in physiological saline solution (0.85% NaCl) using a spectrophotometer [Ultrospec2000 of Pharmacia Biotech (Sweden)]. After adjusting the absorbance at the wavelength of 2 to be equal to 2 MacFarla, inoculate 100 μl of the strain solution diluted in the API liquid medium included in the kit, and then inoculate the strain onto the strip containing 19 sugar substrates. After adding API liquid medium and incubating at 30 ℃ for 72 hours or more, the result of fermentation of each sugar substrate was read and inputted into the program (V4.0; http://apiweb.biomerieux.com ) provided by the manufacturer for physiology and biochemistry. The biochemical properties of various yeast strains stored in the database can be compared and calculated according to their characteristics. Table 1 below shows the results of the API 20C AUX experiment.

Firstly, NK28 isolated from kiwi skin is composed of glucose, galactose, methyl glucopyranoside, maltose, mortar (D-saccharose, sucrose), trehalose (D-trehalose) and melezitose ( D-melezitose) and raffinose (D-raffinose) were available as carbon sources. Saccharomyces cerevisiae La Parisienne, developed as a baker's yeast but also capable of wine fermentation, showed the same results in sugar utilization. On the other hand, Saccharomyces cerevisiae Lv001, which is most commonly used for wine fermentation, was able to use glycerol, which is rarely used by Saccharomyces cerevisiae strains. Lanoside (methyl-aD-glucopyranoside) was not available results.

<Example 5>

Glucose Tolerance Analysis of Saccharomyces cerevisiae NK28 Strains

The strains were inoculated in YPD liquid medium containing 30% or 40% of dextrose as a carbon source, and shaken (220 rpm) for 24 hours at 30 ° C, followed by a spectrophotometer at 600 nm. The absorbance was measured to compare how the strains survived the osmotic shock in the liquid medium containing the high concentration of sugar.

Figure 3 shows the sugar tolerance and maltose utilization of each strain in a graph. The results show that there is no significant difference in the growth rate between NK28 strain and other strains in YPD liquid medium containing 40% glucose, but when 30% glucose is contained, NK28 strain is about 40% higher than Lv001 strain. It showed higher growth rate compared to La Parisienne strain. Each experiment was performed three times and statistically treated as an average value.

<Example 6>

Maltose Utilization Analysis of Saccharomyces cerevisiae NK28 Strains

YPM liquid medium containing 2% maltose instead of glucose in the YPD liquid medium [YPM; 1% yeast extract, 2% peptone, 2% maltose] were inoculated with NK28, Lv001 and Parisienne strains, and shaken at 30 ° C for 24 hours (220 rpm), followed by a spectrophotometer. Absorbance was measured at 600 nm wavelength to compare growth.

As shown in FIG. 3, the NK28 strain in maltose was not significantly different from the La Parisienne strain, but was found to be four times more available than the Lv001 strain used commercially as a wine fermentation strain. Each experiment was performed three times and represented as an average value.

&Lt; Example 7 >

Alcohol Resistance Analysis of Saccharomyces cerevisiae NK28 Strains

YPD liquid exclusion with 6, 7, 8, 9, 10% alcohol added [YPD broth; 1% yeast extract, 1% peptone, 2% peptone, 10% glucose, and then inoculated with the three strains and incubated at 30 ° C for 48 hours (220 rpm), followed by 600 with a spectrophotometer. Absorbance was measured at nm wavelength to determine growth and to compare the resistance to alcohol.

Figure 4 is a test of how well each yeast strain can grow in an environment containing a high concentration of alcohol. In the case of containing 6-7% alcohol, the alcohol resistance of the Lv001 strain was high, but in the case of containing 9-10% alcohol, the NK28 strain was able to grow to a similar extent as the two commercial yeast strains. Each experiment was performed three times and represented as an average value.

&Lt; Example 8 >

Alcohol Fermentation Activity of Saccharomyces cerevisiae NK28 Strains

In order to compare the alcohol fermentation ability of the three types of yeast strains, each strain was inoculated in 5 ml of YPD liquid medium and incubated at 25 ° C. for 72 hours and 96 hours to induce alcohol fermentation. Each fermentation broth was centrifuged (10,000 rpm, 5 minutes) to precipitate the cells, and then the supernatant was taken and filtered with a 0.2 μm ministart syringe filter (Minisart syringe filter, Sartorius Stedim biotech, Germany), followed by high performance liquid chromatography [HPLC, Waters 600S controller, 616 pump, Refractive Index Monitor (Bio-RAD MODEL 1755)] were used to pass through Rezex AlOi organic column (Rezex ROA-Organic Acid H + from Phenomenex (8%)) and washed with purified water. The content of alcohol contained in was measured.

5 is a result of analyzing the alcohol fermentation ability of the yeast strains. Assuming the amount of alcohol produced by Saccharomyces cerevisiae Lv001 commercially available as wine yeast is 100, Saccharomyces cerevisiae NK28 strains were 13% and 5 when fermented for 72 and 96 hours, respectively. It was found to produce more alcohol by%, producing 25% more alcohol in 72 hours and nearly the same amount in 96 hours compared to La Parisienne.

So far I looked at the center of the preferred embodiment for the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

National Institute of Agricultural Science KACC93117 20110207

Claims (8)

Saccharomyces cerevisiae NK28 (accession number KACC 93117P), a new strain for winemaking isolated from kiwi shells. delete The method of claim 1,
The strain is Saccharomyces cerevisiae NK28 ( Saccharomyces) characterized in that using maltose as a carbon source cerevisiae NK28) (accession number KACC 93117P). The method of claim 1,
The strain is Saccharomyces cerevisiae NK28 ( Saccharomyces), characterized in that the fermentation strain wine does not occur during culture cerevisiae NK28) (accession number KACC 93117P). Wine production method, characterized in that using the new wine strain Saccharomyces cerevisiae NK28 (accession number KACC 93117P) separated from the kiwi skin. The method of claim 5,
The wine production method is characterized in that the grape fermented with the starch of cereal wine production method. The method according to claim 6,
The grain is a method of producing wine, characterized in that the rice. Wine produced by the method of any one of claims 5-7.

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