JP2001346573A - Yeast having assimilativity of proline - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a wine having excellent flavor. SOLUTION: This wine is produced by using a yeast belonging to Saccharomyces cerevisiae having abilities for assimilating proline included in a grape juice according to a conventional method for producing the wine.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a yeast strain capable of assimilating proline contained in grape juice, a method for producing wine using the yeast strain, and a wine having a low proline concentration.

[0002]

2. Description of the Related Art Since wine is a favorite item, not only ethanol concentration but also balanced flavor is required. In the wine aroma and body, components derived from the grape juice itself and components produced when yeast ferments grape juice are integrally formed. Therefore, in wine brewing, a commercially available dry yeast having many achievements is often used.

On the other hand, in recent years, diversification of food has progressed, and wines having flavors different from conventional wines have been desired. For this reason, attempts have been made to grow yeasts that produce large amounts of specific esters and organic acids, which are the main components of aroma and body, which greatly affect the quality of wine. For example, using yeast developed by the Japan Brewing Association as a parent strain,
Many reports have been made on the acquisition of mutants that produce various esters as aroma components in large amounts.

[0004] Saccharomyces cerevisiae (Sacchamyces cerevisiae) is the same as yeast used for wine brewing.
It is known about the availability of amino acids of yeast belonging to R. cerevisiae). That is, these yeasts convert amino acids in the medium to amino acid permease (g) when the nitrogen source becomes insufficient in the medium.
general amino acid permeas
e: GAP) for cell cytos
ol), but when the medium becomes supersaturated with carbon dioxide, the intracellular protein is decomposed, the amount of intracellular protein is reduced, and the amount of intracellular amino acids is increased. Mediated incorporation of amino acids is suppressed.

[0005] In addition, the accumulation of ethanol with the progress of fermentation increases the permeability of the plasma membrane, and components such as amino acids pooled in the cells are secreted into the medium. In this way, amino acids in the medium are important nitrogen sources of yeast in the early stage of fermentation in ethanol fermentation, and decrease dramatically, but later in the fermentation, secretion by yeast cells and autolysis of cells ,
It gradually increases and has a great influence on the aroma components of wine.

[0006] Of these amino acids, proline is abundant in wine in the form of free amino acids. The other amino acids, glutamic acid, aspartic acid, lysine, and glycine, which are relatively high, are significantly different from those in the form of polypeptides, but all have a significant effect on the sensory evaluation of wine. ing. In general, it is said that wine yeast cannot assimilate proline because there is no system for transporting proline into cells. Therefore, when the nitrogen content in grape juice is small, it is necessary to add a nitrogen source such as diammonium phosphate to promote fermentation despite the presence of proline.
However, if wine yeast can assimilate proline, it may be possible to produce a sufficient amount of ethanol and produce wine with a different flavor than before, while saving the addition of a nitrogen source, Yeasts with such properties are not yet known.

[0007]

DISCLOSURE OF THE INVENTION The present invention relates to yeast strains having the ability to utilize proline contained in grape juice and belonging to the genus Saccharomyces, and a novel method for producing wine using these yeast strains. And a wine having a low proline concentration.

[0008]

Means for Solving the Problems In order to solve the above problems, the present inventors attempted to screen a yeast having an ability to assimilate proline in grape juice from a wide range of yeast groups. The same genus as the yeast used for brewing, ie, Saccharomyces
yces) was found to have the ability to assimilate proline. Furthermore, when wine is brewed using the yeast, proline in grape juice is assimilated, which cannot be assimilated during fermentation with conventional yeast, and its concentration decreases, and the ethanol concentration is relatively high, The present inventors have found that a wine having a wide and thick taste and a mild flavor can be obtained, and completed the present invention.

That is, the present invention provides (1) a yeast strain having the ability to assimilate proline contained in grape juice and belonging to the genus Saccharomyces, and (2) a yeast strain having an ability to assimilate proline. Is Saccharomyces cerevisiae
es cerevisiae) Y-191 (FERMP
(17) The yeast strain according to (1) or (3) a method for producing wine by subjecting grape juice to ethanol fermentation, wherein the ethanol fermentation is carried out by the presence of the yeast strain according to (1) or (2). (4) wine having a proline concentration of 50 mg / l or less, and (5)
And a wine having a proline concentration of 10 mg / l or less.

[0010] The yeast of the present invention is obtained by fermenting yeast isolated from nature with Delaware seed juice, Concord seed juice and Koshu seed juice, and analyzing the ethanol and amino acids contained in the obtained wine. Can also be obtained by selecting a yeast having a high ethanol concentration and a significantly reduced proline concentration. A typical yeast strain having the ability to assimilate proline includes the Y-191 strain.

The mycological properties of the Y-191 strain are as follows. (1) Morphological properties SPOI agarose medium (potassium acetate 10 g / l, calcium chloride dihydrate 18 mg / ml, magnesium chloride hexahydrate 42 mg / ml, agarose 20 g /
The bacterial strain was applied to 1) and cultured at 30 ° C., and then the ascospores were stained. Ascospores were stained as follows. That is, the cells were fixed on a cover glass, immersed in a 5% chromic acid aqueous solution for 1 to 2 minutes, and then immersed in a 5% malachite green aqueous solution at 80 ° C for 3 to 5 minutes, and then washed in running water.
Furthermore, after immersing in a 0.5% safranin aqueous solution for 10 seconds, it was washed in running water and subjected to microscopic observation. as a result,
One to four spherical ascospores were confirmed.

(2) Physiological properties (2-1) Fermentability of sugar Glucose + sucrose + maltose + galactose-lactose-melibiose-soluble starch-raffinose-(+: positive,-: negative)

(2-2) Assimilation of carbon source Galactose-L-sorbose-sucrose + maltose + cellobiose-trehalose + lactose-melibiose-raffinose + merezitose-soluble starch + D-xylose-L-arabinose-D-arabinose +

D-ribose-L-rhamnose-glycerol + erythritol-ribitol-galactitol-D-mannitol-D-glucitol-salicin-lactic acid-succinic acid-citric acid-inositol- (+: assimilate,-: Does not change)

(2-3) Utilization of nitrate No assimilation

[0016] Based on the above properties, The Yeast
s / a taxonomic study (th
iir revised and enlarged
search), Y-191
The strain is Saccharomyces cerevisiae (Sacchar
omyces cerevisiae). Thus, the present inventors have established this strain as Saccharomyces cerevisiae.
evisiae) Y-191 and deposited with the Ministry of International Trade and Industry, National Institute of Advanced Industrial Science and Technology, Institute of Biotechnology and Industrial Technology on June 6, 2000 as Accession No. FERM P-17886.

The present invention further includes a method for producing wine using the yeast strain of the present invention having the ability to assimilate proline as described above. The production method of the present invention is a method for producing wine by subjecting grape juice to ethanol fermentation, wherein the ethanol fermentation is performed in the presence of the yeast strain of the present invention. There are no particular restrictions on the grape juice used, for example, Koshu, Cabernet Sauvignon, Merlot, Malbec, Pinot Noir, Zinfandel,
Schiller, Chardonnay, Riesling, Chenin Blanc, Gewürztraminer, Pinot Blanc, Muscat, Sauvignon Blanc, Semillon, Grenache,
Juices of any varieties such as Muscadell and Alligote can be used. Of these, the use of fruit juice having a relatively low amino acid content, such as Koshu or Riesling, is particularly preferable because the performance of the yeast strain of the present invention is effectively exhibited.

The ethanol fermentation of grape juice can be carried out by a conventional method. Therefore, conventional conditions may be used for the fermentation temperature, the fermentation period, and the like. For example, the fermentation temperature is normal temperature, for example, in the range of 5 to 30 ° C., and the fermentation period is 1 to
It can be in the range of 30 days. However, there is no intention to limit to this. The amount of the yeast strain of the present invention to be added to grape juice can be appropriately determined in consideration of the components of the grape juice, the type of yeast strain, and fermentation conditions.

The proline concentration in the wine obtained in this manner is 50 mg / l or less, especially 10 mg / l or less when the fermentation period is prolonged, and the wine has a mild taste with breadth, thickness and spread in taste. The wine obtained by ethanol fermentation can be used as a product after appropriate post-treatment if necessary.

[0020]

EXAMPLES Hereinafter, the present invention will be described more specifically with reference to Examples, but it is not intended that the present invention be limited to these Examples.

Example 1 Glucose 10%, malt extract 0.3%, peptone 0.1%
Saccharomyces cerevisiae was placed in an L-shaped tube containing 10 ml of YM medium consisting of 3%, yeast extract 0.3%, and pH 4.0.
e) One loopful of Y-191 strain (FERM P-17886) was inoculated, and cultured with gentle shaking at 28 ° C. for 24 hours using a reciprocating shaker. The culture solution obtained in this way is grape juice (Delaware seed, sugar content of 20.3 ° B
(adjusted to rix) 10 ml was added to 100 ml, and the mixture was further cultured at 20 ° C. for 48 hours to prepare a sake mother. Then, 10% of the sake mother was added to 1000 ml of the grape juice and fermented at 20 ° C. for 6 days to produce wine. As controls, wine yeast, Saccharomyces
Cerevisiae (Saccharomyces cerev)
isiae) Using Zymoflor VL3 (manufactured by Ralvin), wine was produced in the same process as above. The amino acid content in the wine at the end of the fermentation was analyzed using high performance liquid chromatography (HPLC: LC-6A amino acid analysis system (manufactured by Shimadzu Corporation)).

The results are shown in Table 1.

[Table 1]

As shown in Table 1, the content of proline in wine produced using the Y-191 strain was 4.3 mg.
/ Ml, which is remarkably low, indicating that the Y-191 strain has high proline utilization. Also, Y-19
It is presumed that all Y-191 strains, including arginine, which had the highest content among wines produced using one strain, could assimilate amino acids.

Next, Table 2 shows the results of a general analysis according to the analysis method prescribed by the NTA.

[Table 2]

As shown in Table 2, it is clear that good wine can be produced by using the Y-191 strain according to the conventional wine production method. In particular, it was revealed that the amount of ethanol produced was higher than that of the control group, and that the yeast was an excellent yeast having high ethanol fermentation ability.

Example 2 Five males were subjected to a sensory evaluation of the wine prepared in Example 1 and produced using the Y-191 strain. As a control, Zymoflore also prepared in Example 1
When the wine produced using the VL3 strain was used for comparison, the wine of the Y-191 strain was found to be Zymoflor.
e Compared to VL3 wines, 5 people have width and thickness
Two people with mild taste and one person with thickness were evaluated.

Example 3 Glucose 10%, malt extract 0.3%, peptone 0.1%
Saccharomyces cerevisiae was placed in an L-shaped tube containing 10 ml of YM medium consisting of 3%, yeast extract 0.3%, and pH 4.0.
e) One loopful of Y-191 strain (FERM P-17886) was inoculated, and cultured with gentle shaking at 28 ° C. for 24 hours using a reciprocating shaker. The culture solution obtained in this way is mixed with grape juice (Koshu seed, sugar content of 17.4 ° Brix).
10% to 100 ml, and 2 more
Culture was performed at 0 ° C. for 48 hours to obtain a sake mother. Then, 10% of sake mother was added to 1000 ml of the grape juice and fermented at 20 ° C. for 8 days to produce wine.

Table 3 shows the changes in free amino acids during the fermentation period.

[Table 3]

The sampling was performed on the first grape juice, on the third day, (early stage of fermentation: when the consumption of sugar contained in the grape juice was 50% or less), on the fourth day, (mid and late stage of fermentation: 50% to 90% of sugar consumed in grape juice
%) And on day 8 (at the end of fermentation). Y-1
The 91 strain incorporates amino acids other than proline into the cells at the beginning of fermentation and assimilates the cells, and subsequently takes up and assimilates the remaining proline from the latter half of the fermentation to the end of the fermentation. Therefore, it is considered that the Y-191 strain can utilize all free amino acids contained in grape juice.

Table 4 shows the results of the general analysis of the obtained wine according to the analysis method prescribed by the National Tax Agency.

[Table 4]

As shown in Table 4, the use of the Y-191 strain makes it possible to use a grape juice of Koshu species, which is considered to have a relatively low nitrogen content, without adding a nitrogen source such as diammonium phosphate. Wine can be produced.

Claims (5)

[Claims] 1. Saccharomyces having the ability to assimilate proline contained in grape juice.
es) A yeast strain belonging to the genus. 2. A yeast strain capable of assimilating proline is Saccharomyces cerevisiae.
ces cerevisiae) Y-191 (FERM)
P-17886). 3. A method for producing wine by subjecting grape juice to ethanol fermentation, wherein the ethanol fermentation is performed.
Alternatively, a production method performed in the presence of the yeast strain according to claim 2. 4. A wine having a proline concentration of 50 mg / l or less. 5. A wine having a proline concentration of 10 mg / l or less.