KR102024197B1 - Liquid phase yeast culture medium for liquor and production method thereof - Google Patents

Abstract

The present invention relates to a yeast culture medium for liquor in liquid form and a method for preparing the same. Specifically, the present invention has a high growth rate of liquid yeast cultured in a medium prepared by adding a saccharified solution of 8 ° BX sugar, 1% yeast extract and 2% peptone, and adds trehalose or skim milk. By storing at room temperature to confirm that the survival rate is high until 10 weeks after the start of storage, the composition and method according to the present invention can be usefully used to prepare and store the liquid yeast.

Description

LIQUID PHASE YEAST CULTURE MEDIUM FOR LIQUOR AND PRODUCTION METHOD THEREOF}

The present invention relates to a yeast culture medium for liquor in liquid form and a method for preparing the same.

Liquor is a representative fermented food that combines the civilization and history of mankind.It is not clear when alcohol has been, but from the primitive natural fermentation wine, fruit wine in the hunting season, milk liquor in the nomadic age, and grain from the agricultural age. The traditional wine is produced by local sake according to the climate, climate and habits of the world.

The origin of Korean liquor is a legend about the birth myth of Goguryeo Jumong. In addition, there are records of liquor in the literature such as Samguk Sagi, Samguk-Yu, Goryeo Do-Kyung, and Jimin Magic, and it is possible to guess that there have been various liquors since the Three Kingdoms period. During the Joseon Dynasty, brewery technology was advanced, and the Chinese liquor, a supplementary method of dividing raw material purchases, was developed.In addition, the use of glutinous rice was increased in non-glutinous rice, and more than 360 kinds of liquor were produced by the end of the Joseon Dynasty. . However, as the tax law was promulgated during the Japanese colonial period, the traditional liquor that was made for each house was forced to be made into a secret wine, and even the liquor crackdown was reinforced as the liquor crackdown was strengthened. In the 1980s, the use of rice in Takju was allowed as part of efforts to increase rice consumption due to the overproduction and economic margin of rice. Recently, the government has promoted the globalization of traditional liquor industry to promote rice consumption and inherit the traditional culture. Trying hard.

A strain commonly used in the fermentation liquor and the like my process (Saccharomyces) genus Bacillus (Bacillus) genus Lactobacillus bacteria (Lactobacillus) as in Aspergillus (Aspergillus), A saccharide. Saccharomyces cerevisiae is a representative yeast belonging to the asymptomatic fungi, and most of the yeasts used for brewing, including baker's yeast or brewery yeast, are said to be related to this species. Can be.

Recently, as consumer needs for alcoholic beverages diversify and the tendency to find high quality alcoholic beverages increases, the need for excellent seed development is required. Accordingly, there is also a need to develop a method for producing a medium capable of producing a large amount of seed and a method for long term storage of the seed.

Many researchers are working to improve the taste and quality of Takju. However, the existing researches related to Takju mainly consisted of raw materials of Takju and the types of yeast, and few studies on fermentation related to Takju were conducted (KOREAN J. FOOD SCI.TECHNOL.Vol. 42, No. 1, pp. 56-62, 2010).

Thus, the inventors of the present invention while preparing a composition that can mass-produce liquid yeast in a convenient form and using it to find a method for long-term preservation of the liquid form of yeast, 8 ° BX sugar content, Liquid yeast cultured in a medium prepared by adding 1% yeast extract and 2% peptone had high growth rate, and when it was stored at room temperature by adding trehalose or skim milk, the survival rate was maintained until 10 weeks after the start of storage. By confirming that, this invention was completed.

It is an object of the present invention to provide a composition for preparing yeast in a liquid form comprising a saccharification solution, yeast extract and peptone having a sugar content of 1 to 11 brix, and a method for preparing a yeast in liquid form using the same.

Another object of the present invention is to provide a yeast in liquid form prepared by the above method.

In order to achieve the above object, the present invention provides a composition for producing a liquid yeast comprising a saccharification solution, yeast extract and peptone having a sugar of 1 to 11 brix.

In addition, the present invention comprises the steps of 1) preparing a saccharification solution by adding sake and water to rice gourd and malted kodudu; 2) adjusting the sugar content of the saccharified solution to 1 to 11 brix; 3) preparing a culture medium by adding yeast extract and peptone to the saccharified solution of step 2); And 4) provides a method for producing a liquid yeast comprising the step of inoculating the yeast in the culture medium of step 3).

In addition, the present invention provides a yeast in liquid form prepared by the above method.

The present invention has a high growth rate of liquid yeast cultured in a medium prepared by adding a saccharified solution of 8 ° BX sugar content, 1% yeast extract and 2% peptone, and adding trehalose or skim milk at room temperature. By confirming that the survival rate is kept high until 10 weeks after storage, the compositions and methods according to the present invention can be usefully used for preparing and storing liquid yeast.

1 is a saccharification liquid (D2) having a sugar of 2 ° BX, a saccharifying liquid (D4) having a sugar of 4 ° BX, a saccharifying liquid (D6) having a sugar of 6 ° BX, and a saccharifying liquid (D8) having a sugar of 8 ° BX. And it is a graph comparing the growth of yeast according to the sugar content of the saccharified solution (D10) having a sugar content of 10 ° BX.
2 is a diagram illustrating a yeast manufacturing process in a liquid form.
Figure 3 is a graph comparing the survival rate of the liquid form yeast according to the storage temperature (4 ℃, 25 ℃).

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for producing yeast in a liquid form comprising a saccharification solution, yeast extract and peptone having a sugar content of 1 to 11 Brix.

The rice saccharification liquid may be prepared by saccharifying after adding rice alnuruk and water to the gourd rice. The saccharification may be performed at 35 to 45 ° C, specifically at 37 to 43 ° C, more specifically at 38 to 42 ° C. In one embodiment of the present invention, the saccharification may be performed at 40 ℃. In addition, the saccharification may be performed for 2 to 5 days, specifically for 2 to 4 days. In one embodiment of the present invention, the saccharification may be performed for 3 days. At the time of glycosylation, if the temperature is too high, degeneration of the enzyme occurs, and if it is too low, the enzyme activity may be slowed and saccharification may not be performed properly.

The sugar content of the saccharification liquid may be 1 to 11 brix, specifically, 3 to 11 brix, more specifically, 5 to 10 brix. If the sugar content of the saccharification liquid, a nutrient source of yeast is too low, the growth of yeast is reduced due to lack of nutrients, and if too high, alcohol may be produced to decrease the growth of yeast.

The mixing ratio of the saccharified solution, yeast extract and peptone is 3 to 1.5: 1: 3 to 1.5, specifically 2.5 to 1.5: 1: 2.5 to 1.5, more specifically 2.2 to 1.8: 1: 2.2 to 1.8 days Can be. In one embodiment of the present invention, the mixing ratio of the saccharification solution, yeast extract and peptone may be 2: 1: 2. If the yeast extract and peptone are outside the above range, the yeast may grow too much.

The composition may further comprise a preservative. The preservative may be any one or more selected from the group consisting of α-lactose, trehalose and scheme milk. In one embodiment of the invention, the preservative may be α-lactose, trehalose or skim milk.

The yeast is selected from the group consisting of Saccharomyces sp., Wicherhamomyces sp., Pichia sp. And Hansenia spora sp. It may be any one or more. In one embodiment of the invention, the yeast may be of the genus Saccharomyces.

In a specific embodiment of the present invention, the present inventors immersed and dehydrated the washed non-glutinous rice and steamed by adding rice when steam began to rise. After the rice was prepared, saccharified by adding grain malt and water. Thereafter, the sugar content of the saccharified solution was adjusted to 8 ° BX and 1% yeast extract and 2% peptone were added to prepare a composition for preparing yeast in liquid form.

In addition, the present invention comprises the steps of 1) preparing a saccharification solution by adding sake and water to rice gourd and malted kodudu; 2) adjusting the sugar content of the saccharified solution to 1 to 11 brix; 3) preparing a culture medium by adding yeast extract and peptone to the saccharified solution of step 2); And 4) provides a method for producing a liquid yeast comprising the step of inoculating the yeast in the culture medium of step 3).

The gourd rice of step 1) may be prepared by washing rice, immersing, dehydrating and cooking. Immersion can be carried out for 1 to 3 hours, specifically for 1 hour 30 minutes to 2 hours 30 minutes from the start of the washing. In one embodiment of the present invention, immersion can be performed for 2 hours. Dehydration may be performed for 30 minutes to 2 hours, specifically 30 minutes to 1 hour 30 minutes. In one embodiment of the present invention, dehydration may be performed for 1 hour. The cooking may be carried out for 30 minutes to 2 hours, specifically for 30 minutes to 1 hour 30 minutes. In one embodiment of the present invention, the cooker may be performed for 1 hour.

The saccharification of step 1) may be carried out at 35 to 45 ℃, specifically at 37 to 43 ℃, more specifically at 38 to 42 ℃. In one embodiment of the present invention, the saccharification may be performed at 40 ℃. In addition, the saccharification may be performed for 2 to 5 days, specifically for 2 to 4 days. In one embodiment of the present invention, the saccharification may be performed for 3 days.

The sugar content of the saccharification solution of step 2) may be 1 to 11 brix, specifically, 3 to 11 brix, more specifically, 5 to 10 brix.

The mixing ratio of the saccharified solution, yeast extract and peptone is 3 to 1.5: 1: 3 to 1.5, specifically 2.5 to 1.5: 1: 2.5 to 1.5, more specifically 2.2 to 1.8: 1: 2.2 to 1.8 days Can be. In one embodiment of the present invention, the mixing ratio of the saccharification solution, yeast extract and peptone may be 2: 1: 2.

The composition may further comprise a preservative. The preservative may be any one or more selected from the group consisting of α-lactose, trehalose and scheme milk. In one embodiment of the invention, the preservative may be α-lactose, trehalose or skim milk.

The yeast is selected from the group consisting of Saccharomyces sp., Wicherhamomyces sp., Pichia sp. And Hansenia spora sp. It may be any one or more. In one embodiment of the invention, the yeast may be of the genus Saccharomyces.

In a specific embodiment of the present invention, the inventors steamed non-glutinous rice, immersed, dehydrated, and steamed rice when steam began to rise. After the rice was prepared, saccharified by adding grain malt and water. Thereafter, the sugar content of the saccharified solution was adjusted to 8 ° BX and 1% yeast extract and 2% peptone were added to prepare a composition for preparing yeast in liquid form. The composition was inoculated with yeast and shaken to prepare a yeast in liquid form (see FIG. 2).

In addition, the growth of the liquid form yeast prepared by the above method is excellent (see Tables 1 to 3 and FIG. 1), and when treated with trehalose or skim milk with a preservative, the liquid form yeast survived for a long time when stored at room temperature. 5), the liquid form yeast prepared by the above method was confirmed that E. coli was not detected even after long-term storage, so that contamination by other microorganisms did not occur.

Therefore, the present invention can be usefully used for preparing and storing liquid yeast.

The present invention also provides yeast in liquid form prepared by the process of the invention.

The method comprises the steps of 1) adding rice aluruk and water to the soybean rice and then saccharifying to prepare rice saccharified liquid; 2) adjusting the sugar content of the saccharified solution to 1 to 11 brix; 3) preparing a culture medium by adding yeast extract and peptone to the saccharified solution of step 2); And 4) it may be a method for producing a liquid yeast comprising the step of inoculating the yeast in the culture medium of step 3). The method may have the features as described above.

In one example, the saccharification of step 1) may be carried out at 35 to 45 ℃, specifically at 37 to 43 ℃, more specifically at 38 to 42 ℃. In addition, the saccharification may be performed for 2 to 5 days, specifically for 2 to 4 days.

The sugar content of the saccharification solution of step 2) may be 1 to 11 brix, specifically, 3 to 11 brix, more specifically, 5 to 10 brix.

The mixing ratio of the saccharified solution, yeast extract and peptone is 3 to 1.5: 1: 3 to 1.5, specifically 2.5 to 1.5: 1: 2.5 to 1.5, more specifically 2.2 to 1.8: 1: 2.2 to 1.8 days Can be.

The composition may further comprise a preservative. The preservative may be any one or more selected from the group consisting of α-lactose, trehalose and scheme milk.

In a specific embodiment of the present invention, the inventors steamed non-glutinous rice, immersed, dehydrated, and steamed rice when steam began to rise. After making the gourd rice with the increased steamed rice, it was saccharified by adding grain malt and water. Thereafter, the sugar content of the saccharified solution was adjusted to 8 ° BX and 1% yeast extract and 2% peptone were added to prepare a composition for preparing yeast in liquid form. Yeast ( Saccharomyces cerevisiae Y297) was inoculated to the composition and shaken to prepare a yeast in liquid form (see Fig. 2).

In addition, the growth of the liquid form yeast prepared by the above method is excellent (see Tables 1 to 3 and FIG. 1), and when treated with trehalose or skim milk with a preservative, the liquid form yeast survived for a long time when stored at room temperature. 5), the liquid form yeast prepared by the above method was confirmed that E. coli was not detected even after long-term storage, so that contamination by other microorganisms did not occur.

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are only for illustrating the present invention, and the contents of the present invention are not limited thereto.

Confirmation of medium condition suitable for the production of liquid yeast

<1-1> Growth of Yeast by Addition of Glucose, Saccharified Solution or Molasses

In order to select a medium condition suitable for the production of liquid yeast, the change in viable cell count of liquid yeast was measured by the addition of saccharified liquid or molasses liquid, which was obtained by mixing and increasing glucose and rice.

Specifically, 2% glucose is added to YPD, a synthetic medium containing 1% yeast extract, 2% peptone, and 2% dextrose, or a synthetic medium containing 1% yeast extract and 2% peptone. A medium in which 2% saccharified solution was added to phosphorus YP or 2% molasses solution was added to YP was prepared. 1% of Saccharomyces cerevisiae Y297 preculture was inoculated on the prepared medium, and cultured for 6 days at 30 ° C., and the viable cell count of the liquid form yeast was measured according to the medium composition. Take 10 g of the sample into a sterile bag, add 90 ml of sterile saline solution, dissolve it in a homogenizer, dilute stepwise with sterile saline solution, smear the PCA medium, and count the number of cells in CFU (10 ⁵ / ml). Indicated as.

Comparison of Yeast Growth with Addition of Glucose, Saccharin, or Molasses YPD + Glucose YP + saccharification YP + molasses 110 ± 14.4 136.3 ± 17.6 109.3 ± 25.7

As a result, as shown in Table 1, the growth rate of yeast was the highest in the YP medium to which the saccharified solution was added than the YPD to which glucose was added or the YP to which molasses was added (Table 1).

<1-2> Comparison of Yeast Growth with YP or Inorganic Nutrient Added to Saccharified Solution

In order to select a medium condition suitable for the production of liquid yeast, YP or an inorganic nutrient was added to the saccharified solution, and thus the viable cell number of the liquid yeast was measured.

First, YPD, 2% glycosylated solution, YP containing 2% glycated solution, or 2% glycosylated solution and inorganic nutrients (K ₂ HPO ₄ , KH ₂ PO ₄ and (NH ₄ ) ₂ SO ₄ ) YP medium was prepared. Yeast was inoculated into the prepared medium, and cultured for 6 days at 30 ℃ was measured the number of viable cells of liquid form yeast according to the composition of the medium. The number of viable cells was measured in the same manner as in Example <1-1>, and the number of cells was expressed as CFU (10 ⁵ / ml).

Comparison of Yeast Growth with YP or Inorganic Nutrient Added to Saccharified Solution YPD Saccharification Saccharification + YP Saccharified Amount + Inorganic Nutritional Supplement 27 ± 6.1 CFU 46.7 ± 8.7 139.3 ± 12.4 46.3 ± 9.5

As a result, as shown in Table 2, the growth rate of yeast was the highest in the medium prepared by adding YP to the saccharified solution (Table 2).

<1-3> Growth of yeast according to the sugar content of saccharified liquid

In order to select a medium condition suitable for the production of liquid yeast, in Example <1-2> of the liquid form yeast according to the sugar content of the saccharified solution in the medium prepared by adding YP to the saccharified solution which had the highest growth rate of the yeast The viable cell count was measured.

Specifically, after preparing a saccharified solution having a sugar content of 2, 4, 6, 8, or 10 ° BX, it is added to YP medium, and then inoculated with the yeast, and the saccharified solution is incubated at 30 ° C. for 6 days. The change in viable cell count of liquid yeast was measured according to sugar content. The number of viable cells was measured in the same manner as in Example <1-1>, and the number of cells was expressed as CFU (10 ⁵ / ml).

Comparison of Yeast Growth with Sugar Contents 2 ° BX 4 ° BX 6 ° BX 8 ° BX 10 ° BX 26.7 ± 8.02 95.3 ± 17.62 119.0 ± 16.26 156 ± 6.24 138 ± 1.73

As a result, as shown in Table 3 and Figure 1, the growth rate of yeast was the highest in the medium prepared by adding YP to the saccharification solution of 8 ° BX (Table 3 and Figure 1).

<1-4> Preparation of yeast in liquid form

Based on the results of the above Examples <1-1> to <1-3> to establish the optimum medium conditions for the production of the liquid form of the yeast, by using this to prepare a liquid form of yeast.

Specifically, 6 kg of non-glutinous rice was washed and soaked for 2 hours from the time when the washing was started. After immersion, dehydration was carried out for 1 hour, and when steam began to rise, rice was added and steamed for 1 hour. After the steamed rice was prepared with the steamed steam, 1,080 g of grain nuruk (entry) and 9 L of water were added to saccharify at 40 ° C. for 3 days. Then, the sugar content of the saccharified solution was adjusted to 8 ° BX and 1% yeast extract and 2% peptone were added. The medium was inoculated with 1% yeast ( Saccharomyces cerevisiae Y297) and shaken at 25 rpm for 48 hours at 25 ℃ to prepare a liquid yeast (Fig. 2).

Identification of preservation conditions for long term storage of liquid yeast

<2-1> Comparison of Survival Rate of Liquid Yeast by Storage Temperature

In order to compare the survival rate of liquid form yeast according to storage temperature, the liquid form yeast prepared in Example <1-4> was stored at 4 ° C. at low temperature or 25 ° C. at room temperature, and the number of viable cells of yeast during the storage period. The change was measured. The number of viable cells was measured in the same manner as in Example <1-1>, and the number of cells was expressed as CFU (10 ⁶ / ml).

Comparison of Survival Rate of Liquid Yeast by Storage Temperature storage duration
4 ℃ 25 ℃ CFU (10 ⁵ / ml) Log (CFU / mL) Relative
Survival rate (%) CFU (10 ⁵ / ml) Log (CFU / mL) Relative
Survival rate (%) 0 days 142.3 ± 6.8 8.152 100 163.7 ± 5.2 8.214 100 20 days 76.3 ± 2.4 7.883 96.69 68.7 ± 1.1 7.837 95.41 30 days 14.7 ± 6.6 7.167 87.92 8.2 ± 1.1 6.914 84.17 60 days 12.6 ± 1.5 7.100 87.10 0.4 ± 0.1 5.602 68.20 150 days 2.3 ± 1.1 6.362 78.04 0.0 ± 0.0 0.000 0.00

As a result, as shown in Table 4 and Figure 3, there was almost no difference in survival rate with temperature until 30 days after the start of storage. However, after 30 days, the survival rate was different according to temperature. At 4 ℃, the yeast survival rate was 87.1% and 78.0% at 60 days and 150 days of storage, respectively. At 25 ℃, room temperature, the survival rates of yeast at 60 and 150 days of storage were 68.2% and 0%, respectively. Yeast survival was higher for long term storage (Table 4 and Figure 3).

<2-2> Survival Comparison of Liquid Yeast by Preservative Treatment

In order to compare the survival rate of the liquid form yeast according to the preservative treatment, the liquid form yeast prepared in Example <1-4> was not treated with the preservative, 5% α-lactose, 5% trehalose or 5% Treated with the skim milk (skim milk) and stored at 25 ℃ was measured the number of viable cells of yeast during storage period. The number of viable cells was measured in the same manner as in Example <1-1>, and the number of cells was expressed as CFU (10 ⁴ / ml).

Comparison of Survival Rate of Liquid Yeast by Preservative Treatment storage duration
No treatment α-lactose CFU (10 ⁴ / ml) Log (CFU / mL) Relative
Survival rate (%) CFU (10 ⁴ / ml) Log (CFU / mL) Relative
Survival rate (%) 0 weeks 186.0 ± 1.0 6.27 100 181.1 ± 1.0 6.26 100 2 weeks 131.0 ± 1.0 6.12 97.6 123.3 ± 1.0 6.09 97.3 4 weeks 113.3 ± 5.0 6.05 96.6 98.9 ± 1.1 6.00 95.8 6 Weeks 37.7 ± 1.0 5.58 88.9 55.7 ± 1.0 5.75 91.8 8 Weeks 3.7 ± 1.0 4.57 72.9 20.7 ± 1.0 5.32 84.9 10 Weeks - - - 10.0 ± 0.2 5.00 79.9 15 Weeks 3.3 ± 1.0 4.52 72.1 12.3 ± 1.0 5.33 85.1 storage duration
Trehalose Scheme milk CFU (10 ⁴ / ml) Log (CFU / mL) Relative
Survival rate (%) CFU (10 ⁴ / ml) Log (CFU / mL) Relative
Survival rate (%) 0 weeks 164.6 ± 1.0 6.22 100 170.3 ± 1.0 6.23 100 2 weeks 126.9 ± 1.0 6.10 98.2 135.5 ± 1.0 6.13 98.4 4 weeks 129.5 ± 1.1 6.11 98.3 132.0 ± 1.2 6.12 98.2 6 Weeks 61.6 ± 1.0 5.79 93.1 64.0 ± 1.0 5.81 93.2 8 Weeks 34.6 ± 0.1 5.54 89.1 47.7 ± 0.1 5.68 91.1 10 Weeks 18.8 ± 1.0 5.27 84.8 24.7 ± 1.0 5.39 86.5 15 Weeks 18.7 ± 1.0 5.09 81.9 21.3 ± 1.0 5.27 84.6

As a result, as shown in Table 5, the relative survival rate was more than 95% until the 4th week of storage, and the liquid form yeast treated with skim milk was higher than the liquid form yeast without the preservative at the 8th week of storage. Indicated. At 10 weeks of storage, the liquid form yeast without preservatives was almost killed, but the liquid form yeast treated with trehalose or skim milk showed a survival rate of over 85%, indicating that trehalose or skim milk was useful for room temperature storage of liquid form yeast. It was found (Table 5).

<2-3> Confirmation of E. Coli Contamination in Liquid Yeast

In order to confirm the degree of E. coli contamination according to the storage period of the liquid form yeast, E. coli detection test was carried out while storing the liquid form yeast prepared in Example <1-4> at 25 ° C. The degree of contamination was confirmed.

As a result, E. coli was not detected even at 10 weeks and 15 weeks of storage, it was found that the liquid yeast is not contaminated by other microorganisms.

Claims (11)

1) preparing rice saccharification liquid by adding rice alnuruk and water to the soybean rice and then saccharifying it;
2) adjusting the sugar content of the saccharified solution to 8 brix;
3) preparing a culture medium by adding yeast extract and peptone to the saccharified solution of step 2);
4) inoculating and incubating the yeast in the culture medium of step 3) to prepare a yeast in liquid form; And
5) A process for preparing a liquid yeast having a relative survival rate of at least 80% at 15 weeks, comprising treating trehalose or skim milk with a preservative to the liquid yeast of step 4).
The method of claim 1, wherein the yeast is Saccharomyces sp., Wicherhamomyces sp., Pichia sp. And Hansenia spora sp. At least one selected from the group consisting of, a method for producing a yeast in liquid form.
Yeast in liquid form prepared by the method of claim 1. delete delete delete delete delete delete delete delete

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