KR101860409B1 - Saccharomyces cerevisiae which is a novel yeast - Google Patents

Abstract

The present invention relates to saccharomyces cerevisiae KSD-BH as a novel yeast, which has high fermentation rate and low mortality rate when fermenting an alcoholic drink, can use a sugar selected from a group consisting of glucose, glycerol, galactose, maltose, saccharose, trehalose and raffinose, and can be accessed with the accession number of KACC 93275P.

Description

A new yeast strain, KSD-BH {SACCHAROMYCES CEREVISIA WHICH IS A NOVEL YEAST}, was introduced into Saccharomyces cerevisiae,

The present invention is directed to KSD-BH in Saccharomyces cerevisiae. It is a new yeast isolated from yeast.

Recently, makgeolli using various materials has been developed and consumed (Korean Patent Publication No. 10-2017-0030672, Korean Patent No. 10-1723162, etc.). However, in order to diversify the material and taste, there are various alcohol concentration, fermentation temperature, etc. In this case, the survival rate of the yeast and the fermentation efficiency are often not as good as desired.

Therefore, the inventors of the present invention have found that a novel yeast isolated from yeast while fermenting yeast having a high spectrum of use in the production of makgeolli can ferment low amino acids and thus can realize a clean quality without highly digesting the two kinds of rice. The fermentation rate is high and the spectrum of the fermentation temperature is wide. Therefore, since the fermentation heat generated during the fermentation of alcohol is not required to be lowered to the cooling water, the fermentation stability is maintained and the fermentation stability is maintained The alcohol yield and the productivity can be increased, and the present invention has been completed.

It is an object of the present invention to provide a novel yeast useful for producing makgeolli.

According to an aspect of the present invention,

A saccharide selected from the group consisting of glucose, glycerol, galactose, maltose, saccharose, trehalose and raffinose can be used, which has a high fermentation rate and high mortality rate during fermentation, : Provide KSD-BH in Saccharomyces cerevisiae entrusted with KACC 93275P.

KSD-BH in the saccharomyces cerevisiae of the present invention can be fermented with low amino acid, so that it can be economically advantageous in that it can realize a clean quality without highly digesting of rice for two kinds of dishes, Low fermentation rate and high spectrum of fermentation temperature, it is not necessary to lower the fermentation heat generated by alcohol fermentation to the cooling water, so the process cost is low during fermentation, and fermentation stability is maintained, thereby improving the alcohol yield and productivity .

Fig. 1 shows the result of comparing the standard ITS1-5.8S rRNA-ITS2 base sequence parts contained in the same species and genus as KSD-BH in S. cerevisiae.
Fig. 2 shows a taxonomic phylogenetic tree according to the ITS gene sequence of KSD-BH in S. cerevisiae.
Figure 3 shows the change in alcohol concentration per hour during fermentation at 25 < 0 > C.
Figure 4 shows the change in alcohol concentration per hour during fermentation at 20 < 0 > C.
Figure 5 shows the change in alcohol concentration per hour during fermentation at < RTI ID = 0.0 > 15 C. < / RTI >

According to the present invention,

A sugar selected from the group consisting of glucose, glycerol, galactose, maltose, saccharose, trehalose and raffinose, which has high fermentation efficiency and low mortality rate during fermentation, can be used. : For KSD-BH in Sakaromyces cerevisiae entrusted with KACC 93275P.

The present invention also relates to a food composition to which KSD-BH is added to Saccharomyces cerevisiae.

Also,

1. A method for producing a fermented product, comprising the steps of: mixing rice, water, yeast, and KSD-BH with Saccharomyces cerevisiae of claim 1 to produce a first stage fermentation product; And

And further adding rice, water and yeast to the first-stage fermentation product to produce a second-stage fermentation product by two-stage fermentation.

Hereinafter, the present invention will be described in detail.

Sakaromayses Serebijie KSD -BH

KSD-BH in the saccharomyces cerevisiae of the present invention is a fermentation broth of glucose, glycerol, galactose, maltose, saccharose, trehalose and raffinose, which is separated from the restored traditional yeast by fermentation with high fermentation ratio and low mortality rate A sugar selected from the group consisting of sugar chains can be used, and it has been deposited under accession number: KACC 93275P. In this case, KSD-BH in saccharomyces cerevisiae has a mortality rate of 5% or less at 20 ° C when brewing fermentation with rice as a raw material. In addition, KSD-BH at saccharomyces cerevisiae can have an extinction rate of 10% or less at 15 ° C and an extinction rate of 50% or less at 25 ° C. KSD-BH has a fermentation ratio of 91% or more at 25 ° C, 89% or more at 20 ° C, and 88% or more at 15 ° C in Saccharomyces cerevisiae, and the fermentation ratio is calculated according to the following formula 2.

<Formula 2>

In addition, KSD-BH may be added to saccharomyces cerevisiae of the present invention in the presence of 2-keto-D-gluconate, arabinose, xylose, adonitol, xylitol, inositol, sorbitol, methyl- A sugar selected from the group consisting of glucosamine, cellobiose, lactose, and melitose is not used.

KSD-BH in the above-mentioned Saccharomyces cerevisiae is derived from the yeast.

In addition, KSD-BH in the saccharomyces cerevisiae of the present invention can be fermented with low amino acid, so that it can be economically advantageous in that it can realize a clean quality without highly digesting two kinds of rice, The fermentation temperature is low and the spectrum of the fermentation temperature is wide so that it is not necessary to lower the fermentation heat generated by the alcohol fermentation to the cooling water so that the fermentation stability is maintained and the alcohol yield and productivity can be improved .

Food composition

The present invention relates to a food composition to which KSD-BH is added to Saccharomyces cerevisiae. The type of the food is not limited and may be a general food, a health supplement, a functional food, or the like. Preferably, the food composition is a food composition in which yeast is used. More preferably, the food composition is a mainstream, for example, a fermentation liquor and a distillate liquor such as Yakju, Takju, and sake.

Manufacturing method of makgeolli

The present invention relates to a method for producing a fermented product by mixing KSD-BH with rice, water, koji and the saccharomyces cerevisiae of claim 1, And further comprising adding rice, water and yeast to the first-stage fermentation product to perform a second-stage fermentation to produce a second-stage fermentation product. At this time, it is possible to carry out the method of manufacturing the makgeolli of the present invention by using a general technique of producing makgeolli using rice, water, koji, or the like.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention, and the manner of achieving them, will be apparent from and elucidated with reference to the embodiments described hereinafter in conjunction with the accompanying drawings. It should be understood, however, that the invention is not limited to the disclosed embodiments, but is capable of many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

&Lt; Materials and methods >

The mung bean and glutinous rice used in the manufacture of yeast were purchased from domestic marketed mung bean and domestic glutinous rice. Rice used in making makgeolli is domestic rice.

Experimental Example 1 Isolation of Strain

(1813) and Yangcheon (1837) were reproduced to produce traditional yeast. First, 100 parts by weight of mung bean washed with water was added, and the mixture was cooled with cold water. And 50 parts by weight of glutinous rice was made into powder after being called water. The mung bean and the glutinous rice powder were mixed and put together in a mortar, and then the mixture was packed into a paper box. The paper box was placed on a pine needle and stored at room temperature for 7 days without sunshine. Then, after 2 weeks, it was aired, and at 3 weeks, it was dried in the sun to produce a traditional yeast.

The traditional yeast was sampled and diluted with sterilized distilled water to isolate the strains. The yeasts were selected from the isolated strains, and the fermentation ratio and the alcohol production ability (i.e., brew suitability) of the yeasts were examined. Among them, strains having the best fermentation ratio and alcohol production ability were selected (test results are not described).

<Experimental Example 2> Confirmation by phylogenetic analysis

The nucleotide sequence of the ITS gene was analyzed in order to confirm the species of the strain isolated from the conventional yeast, which was confirmed to have the best brewing suitability in Experimental Example 1 above. The analysis was performed by sequencing using ITS1 and ITS4 primers from Solgent (Daejeon, Korea). Specifically, the ITS gene full-length sequence was assembled using SeqMan software (DNASTAR) and blasted into the NCBI database. And genetically related ITS gene sequences were obtained from GenBank and the nucleotide sequences were compared with BioEdit program.

We also constructed a phylogenetic tree by using the MEGA 3 program and the neighbor-joining method with a bootstrap value based on 1000 replications.

As a result, the nucleotide sequence of the ITS gene was as shown in the following SEQ ID NO: 1 (Table 1), and the standard strains belonging to the genus Saccharomyces or Saccharomyces cerevisiae and the ITS1-5.8S rRNA-ITS2 base sequence As a result, it was confirmed that this strain belonged to Saccharomyces cerevisiae (Fig. 1). The taxonomic classification according to the ITS gene sequence was as shown in Fig. 2 (Fig. 2), and the ITS gene sequence was confirmed to be a novel strain because the ITS gene sequence was quite different from the conventional Saccharomyces cerevisiae. Therefore, it was named KSD-BH in Saccharomyces cerevisiae and deposited with the National Academy of Sciences on January 24, 2017 (Microorganism Accession No .: KACC 93275P).

<Table 1>

&Lt; Experimental Example 3 >

Saccharomyces cerevisiae KSD-BH was assayed for its sugar-using properties using an API 20C AUX kit (Biomerieux, France). As the comparative strains, KSD-SF was used for Saccharomyces cerevisiae and KSD-YP was used for Saccharomyces cerevisiae.

As a result, KSD-BH was able to utilize glucose, glycerol, galactose, maltose, saccharose, trehalose and raffinose in the saccharomyces cerevisiae of the present invention. However, in the saccharomyces cerevisiae of the present invention, KSD-BH is preferably selected from the group consisting of 2-keto-D-gluconate, arabinose, xylose, adonitol, xylitol, inositol, sorbitol, methyl- Glucosamine, Cellobiose, Lactose, and Melitose were not available (Table 2).

<Table 2>

&Lt; Experimental Example 4 >

The alcohol resistance of KSD-BH was evaluated in Saccharomyces cerevisiae. First, cultured yeast strains were inoculated in 30 mL of YPD medium containing ethanol at concentrations of 1, 5, 10, and 15% (v / v) at a concentration of 1.0E + 05 cells / mL, 30 [deg.] C and 150 rpm. After 24 hours of incubation, the culture broth was diluted and the absorbance was measured at 600 nm.

As a result, KSD-BH in the saccharomyces cerevisiae of the present invention was higher in alcohol resistance than other yeasts (Table 3).

<Table 3>

&Lt; Experimental Example 5 >

The saccharomyces cerevisiae was evaluated for glucose tolerance of KSD-BH. First, the cultured larvae were inoculated into 30 mL of YPD medium containing glucose at concentrations of 2, 10, 20, 30, and 40% (w / v), respectively, at a concentration of 1.0E + 05 cells / mL And then cultured at 30 ° C and 150 rpm. After 24 hours of incubation, the culture medium was collected, diluted appropriately, and absorbance was measured at 600 nm.

As a result, KSD-BH in Saccharomyces cerevisiae of the present invention was more resistant than other yeasts (Table 4).

<Table 4>

&Lt; Experimental Example 6 >

900 g of rice, 1350 mL of water, 8.6 g of yeast and yeast culture were mixed. At this time, the mixture was mixed at a concentration of 6.5E + 07 cells / mL per ml of the syringe, and a one-stage immersion was performed. The fermentation was carried out at 15 ° C, 20 ° C and 25 ° C for 1 day to perform one stage fermentation.

After the first stage fermentation, 2,100 g of rice, 3,150 mL of water and 20.1 g of yeast were further added, followed by two stages of immersion, followed by two stages of fermentation at 15 ° C, 20 ° C and 25 ° C. At this time, the temperature was the same for the first stage fermentation and the second stage fermentation, and each experiment was carried out by using four kinds of yeasts (S. cerevisiae KSD-BH, S. cerevisiae KSD-YP, S. cerevisiae KSD-SF, S. cerevisiae KSD-SC).

The yeast kill rate during fermentation was calculated according to the following formula 1. Alcohol concentration and fermentation rate were measured and calculated at fermentation temperature of 25 ℃ and 20 ℃ for 8 days and fermentation was performed at 15 ℃ for 16 days. The fermentation ratio was calculated according to the following formula (2). After the fermentation, the fermented syrup was filtered with a filter paper and neutralized with 0.1 N NaOH. Then, 5 mL of neutral formalin solution was added to the mixture, which was then titrated with 0.1 N NaOH solution to obtain 0.1 N NaOH.

<Formula 1>

<Formula 2>

As a result, the amino acid degree of brewing using KSD-BH in Saccharomyces cerevisiae of the present invention was lower than that of other yeasts (Table 5), and yeast mortality was low (Table 6). On the other hand, KSD-BH in the saccharomyces cerevisiae had high alcohol production efficiency and high fermentation rate in brewing. The alcohol fermentation temperature showed a high spectrum, especially the higher the fermentation temperature, the higher the alcohol concentration. Therefore, it was judged that high-concentration alcohol fermentation would be possible at 25 캜 even without a separate cooling device (Tables 7 to 10, Figs. 3 to 5). Therefore, KSD-BH was determined to be suitable for use in the manufacture of sake.

<Table 5> Amino acid composition during fermentation according to fermentation temperature and yeast type

<Table 6> Yeast kill rate by fermentation temperature and yeast type

<Table 7> Alcohol content and fermentation rate of fermented product according to temperature and yeast type

<Table 8> Changes in Alcohol Concentration with Fermentation Time at 25 ℃

<Table 9> Changes in Alcohol Concentration with Fermentation Time at 20 ℃

<Table 10> Change of alcohol concentration according to fermentation time at 15 ℃

<Experimental Example 7> Sensory properties

In the Experimental Example 6, a sensory test was performed on the final fermentation broth of each brewing yeast fermented and manufactured at 20 ° C. First, 15 trained test personnel are selected and the taste, incense, and overall preference are assessed through a 9-point scale (1-not good, 3-bad, 5-normal, 7-good, Respectively. The results are shown in Table 11 below.

<Table 11> Example 6, sensory evaluation of the fermentation broth of 20 ° C for brewing yeast

As shown in Table 11, KSD-BH in S. cerevisia showed similar or high preference in flavor, taste and general preference in all items. It can be confirmed that when brewing with S. cerevisiae using KSD-BH, it is possible to impart sensory qualities such as taste and flavor.

Agricultural Biotechnology Research Institute KACC93275 20170124

<110> KOOKSOONDANG CO., LTD <120> SACCHAROMYCES CEREVISIAE WHICH IS A NOVEL YEAST <130> KNP170011 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 727 <212> DNA <213> Saccharomyces cerevisiae <400> 1 atttttttgt tttggcaaga gcatgagagc ttttactggg caagaagaca agagatggag 60 agtccagccg ggcctgcgct taagtgcgcg gtcttgctag gcttgtaagt ttctttcttg 120 ctattccaaa cggtgagaga tttctgtgct tttgttatag gacaattaaa accgtttcaa 180 tacaacacac tgtggagttt tcatatcttt gcaacttttt ctttgggcat tcgagcaatc 240 ggggcccaga ggtaacaaac acaaacaatt ttatttattc attaaatttt tgtcaaaaac 300 aagaattttc gtaactggaa attttaaaat attaaaaact ttcaacaacg gatctcttgg 360 ttctcgcatc gatgaagaac gcagcgaaat gcgatacgta atgtgaattg cagaattccg 420 tgaatcatcg aatctttgaa cgcacattgc gccccttggt attccagggg gcatgcctgt 480 ttgagcgtca tttccttctc aaacattctg tttggtagtg agtgatactc tttggagtta 540 acttgaaatt gctggccttt tcattggatg tttttttttc caaagagagg tttctctgcg 600 tgcttgaggt ataatgcaag tacggtcgtt ttaggtttta ccaactgcgg ctaatctttt 660 ttatactgag cgtattggaa cgttatcgat aagaagagag cgtctaggcg aacaatgttc 720 ttaaagt 727

Claims (5)

It is possible to use one or more sugars and glycerol selected from the group consisting of glucose, galactose, maltose, saccharose, trehalose and raffinose, can not use meleletose, And the fermentation ratio is calculated according to the following formula (2).

<Formula 2>

Accession number: KSD-BH in Saccharomyces cerevisiae entrusted with KACC 93275P.
The method according to claim 1,
And KSD-BH in a saccharomyces cerevisiae characterized in that rice is used as a raw material at 15 to 20 ° C and has an mortality rate of 10% or less upon fermentation under brewing.
The method according to claim 1,
And KSD-BH in a saccharomyces cerevisiae having an amino acid degree of 1.5 or less when brewing fermentation with rice as a raw material at 15 to 20 ° C.
delete A food composition comprising KSD-BH added to the saccharomyces cerevisiae of claim 1.

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