JP3260919B2 - Food and beverage manufacturing method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing foods and drinks such as alcoholic beverages, bread, fermented seasonings (also called brewed seasonings) using mutant yeast belonging to the genus Saccharomyces.

[0002]

2. Description of the Related Art As a method for improving the flavor of foods and drinks using Saccharomyces cerevisiae, for example, a method using an azaleucine-resistant strain to increase isobutyl alcohol and isoamyl alcohol in foods and drinks (Japanese Unexamined Patent Publication No. 257423), a method using glyphosate-resistant strain, thienylalanine-resistant strain, thiazolealanine-resistant strain, chloroalanine-resistant strain or propargylglycine-resistant strain (JP-A-3-7579), and aminoethylcysteine-resistant strain Method (JP-A-3-58
733), a method using an azetidine carboxylate-resistant strain (Japanese Patent Application Laid-Open No. 4-91782), and a method using a fluorophenylalanine-resistant strain to increase β-phenethyl alcohol (Japanese Patent Application Laid-Open No. 2-92265).
Japanese Patent Application Laid-Open No. 4-252135) and a method using a thiisoleucine-resistant strain to increase active amyl alcohol and n-propyl alcohol.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel mutation capable of producing a larger amount of aroma components such as higher alcohols useful as aroma components of foods and drinks, especially active amyl alcohol and n-propyl alcohol. An object of the present invention is to provide a yeast and a method for producing a fragrant food or drink using the yeast.

[0004]

SUMMARY OF THE INVENTION The present invention relates to a method for producing food or drink using yeast, which comprises using a yeast belonging to the genus Saccharomyces and having methylthreonine resistance, thereby producing a highly aromatic alcoholic beverage, bread, and fermented seasoning. The present invention relates to a method for producing food and drink such as food. Examples of alcoholic beverages include shochu, whiskey, wine, sake, and the like. Examples of breads include bread and confectionery bread. Examples of fermented seasonings (alcoholic beverages) include mirin and sake-style seasonings. .

[0005] As the yeast used in the present invention, any yeast belonging to the genus Saccharomyces and having methylthreonine resistance can be used. Methylthreonine-resistant strains can be obtained by mutating commercially available yeast (eg, sake yeast, wine yeast, beer yeast, whiskey yeast, shochu yeast, baker's yeast, etc.) by a known mutagenesis method, for example, by irradiating ultraviolet rays, radiation, or the like. -Methyl-N'-nitro-N
-Nitrosoguanidine, ethyl methanesulfonate and the like can be obtained by suitably using a method of contacting a drug.

For example, a mutagenized yeast is applied to an agar plate medium containing methylthreonine at such a concentration that the parent strain cannot grow, the grown colonies are separated, and the amount of aroma components produced from the separated colonies is increased. By selecting a mutant strain, a target mutant strain can be obtained. Specific examples of the strain used in the present invention include Saccharomyces cerevisi ( Saccharomyces cerevisi).
ae ) MT31 strain (hereinafter referred to as MT31 strain).

Next, a method for obtaining the MT31 strain will be specifically described. Saccharomyces cerevisiae Japan Brewing Association No. 7 yeast (hereinafter referred to as K7 strain) cultured at 30 ° C. for 16 hours in a YPD medium (1% yeast extract, 2% peptone, 2% glucose, pH 6.0) was 0.2M. Phosphate buffer (p
H8.0) to a cell concentration of 10 ⁶ cells / ml.

Then, ethyl methanesulfonate is added to the suspension to a final concentration of 30 μl / ml,
After standing at 30 ° C. for 60 minutes, the cells are collected by centrifugation. After washing the cells with a 5% aqueous sodium thiosulfate solution, the cells are further washed with a phosphate buffer (pH 6.0). The cells were cultured at 30 ° C. for 10 days in a plate medium (Bacto yeast nitrogen base 0.67%, glycerol 2%, agar 2%, manufactured by Difco) containing 2 mg / ml of orthomethylthreonine, and appeared. A colony is separated to obtain a methylthreonine-resistant strain. For selection of superior mutants, the cells were cultured in YPD medium at 30 ° C. for 3 days, and then the odor components contained in the culture solution were quantified. It was named threonine resistant strain MT31.

The MT31 strain was deposited on July 16, 1993, with the Institute of Biotechnology and Industrial Technology, FERMBP-4364, based on the Budapest Treaty. Next, the sensitivity of the above mutant strain and its parent strain to the drug was examined. Orthomethylthreonine was mixed with 0.67% of Bacto yeast nitrogen base (manufactured by Difco), 2% of glycerol, and 2% of agar to each of the concentrations shown in Table 1 to obtain an agar plate medium. After inoculating this culture medium with the mutant strain and its parent strain,
The growth after day was observed. The results are shown in Table 1.

[0010]

[Table 1]

The method for producing alcoholic beverages, fermented seasonings and bread using the mutant strain and MT31 strain can be carried out by conventional production methods. For example, production of alcoholic beverages and fermented seasonings will be described. The carbon source is saccharified with enzyme or koji, and yeast is added for fermentation. In the production of sake and shochu, generally, a part of the carbon source is charged at the start of fermentation, and the rest is added as the fermentation progresses.

As the carbon source, any carbohydrate and starch can be used, and raw materials specified in the Sake Tax Law and the Sake Tax Law enforcement order depending on the type of alcoholic beverage to be used are used. For example, fruits such as grapes, Molasses, glucose, potatoes, buckwheat, rice, barley, bubble, corn, corn, fly, cane, and koji of these are used. The saccharification enzymes, generally those included in malt, which produce the Aspergillus, such as filamentous fungi Aspergillus oryzae (Aspergillus oryzae), alpha chromatography amylase, glucoamylase, enzyme preparations such as protease can be used.

As the yeast, those belonging to the genus Saccharomyces and having methylthreonine resistance can be used alone or together with other yeasts. When fermentation is performed using fruits such as grape, molasses, glucose or the like, single fermentation in which yeast is directly added for fermentation is performed. When cereals such as potatoes, rice, wheat, buckwheat, and corn are used, parallel double fermentation is performed in which starch, which is cereals, is first decomposed into saccharides by a saccharifying enzyme, and then yeast is added for fermentation.

The fermentation is carried out, for example, at pH 3.5-5.0, 5-
Perform at 25 ° C. for 10-30 days. Next, in order to make the moromi produced by fermentation of yeast into an alcoholic beverage, various methods are employed depending on the desired alcoholic beverage. For example, in the case of sake, wine, etc., after completion of the moromi fermentation, the unrefined liquor is obtained by separating the fermentation residue, yeast cells and the like by squeezing filtration or the like. For distilled spirits (shochu, whiskey, etc.)
Moromi is distilled using a simple distillation machine or the like to obtain an original sake. The fermented seasoning is obtained by performing a treatment of adding salt, acetic acid, and the like to moromi before the end of fermentation (a non-drinking treatment prescribed by the Liquor Tax Law) and then filtering.

Next, the production of bread will be described. The yeast is prepared in a normal medium containing a carbon source, a nitrogen source, an inorganic substance, an amino acid, a vitamin, etc. under aerobic conditions at a temperature of 26 to 33 ° C.
Culture is performed while adjusting to H4 to 6, and the cells are collected and washed to obtain yeast cells suitable for bread production. As the carbon source in the medium, various carbohydrates such as glucose, sucrose, starch hydrolyzate, molasses and the like can be used. In particular, molasses is preferably used.

As the nitrogen source, ammonia, ammonium chloride, ammonium sulfate, ammonium carbonate, ammonium acetate, urea, yeast extract, corn steep liquor and the like are used. Other inorganic substances, amino acids, vitamins, and the like can be added as needed. Culture is
Fed-batch culture is appropriate. When bread is manufactured using the straight method, for example, it is performed as follows.

Wheat flour, sugar, the yeast cells obtained above,
Water is added to the raw material mainly composed of shortening and the like, kneaded, and then fermented at 25 to 35 ° C. for 30 to 150 minutes. Then, the dough is divided and left at 15 to 35 ° C. for 10 to 30 minutes (bench time). After standing, the dough is molded, put in a mold, and the dough expands to a certain height.
Perform final fermentation at 45 ° C. 10-4 at 180-240 ° C
Bake for 0 minutes to make bread.

When bread is manufactured using the sponge method, it is performed, for example, as follows. Water is added to the flour, the above-obtained medium seed material mainly containing yeast cells and the like, and kneaded, and then kneaded.
Fermentation (medium seed fermentation) at 35 ° C for 2 to 5 hours. To this fermented product, a main kneading raw material mainly composed of flour, sugar, shortening and the like and water are added and kneaded to obtain a dough. The dough is usually left at 25 to 35 ° C. for 10 to 40 minutes (floor time). Then, the dough is divided, and then
Leave for 0 minutes (bench time). The dough is molded, put into a mold, and subjected to final fermentation at 35 to 45 ° C until the dough expands to a certain height, and then baked at 180 to 240 ° C for 10 to 30 minutes to produce bread.

[0019]

Examples are shown below. Embodiment 1 FIG. Production of Sake K7 strain and MT31 strain were used as yeast. Moromi was prepared as shown in Table 2.

[0020]

[Table 2]

Moromi management: Moromi temperature is 16 ° C from garnishing, dancing and arranging, 14 ° C from arranging to closing, and 13 ° C after arranging,
Thereafter, the temperature is raised by 1 ° C. every day, and after the start of falling bubbles,
Conversely, the temperature was lowered by 1 ° C. per day to 10 ° C. Three days after the garnishing, the middle garment was prepared, and one day after that, the garment was charged, and the upper tank was placed 19 days after the garment was charged.

Table 3 shows the components of the brewed sake.

[0023]

[Table 3]

Embodiment 2 FIG. Production of barley shochu K7 strain and MT31 strain were used as yeast. Moromi was prepared as shown in Table 4 and rice koji was used for the primary preparation and barley having a degree of whitening of 75% was used for the secondary preparation.

[0025]

[Table 4]

Moromi management: Moromi temperature is 20 ° C.
And Three days after the first charge, the second charge, and one more
Distillation was performed 0 days later. Table 5 shows the flavor components of the shochu obtained by distillation.

[0027]

[Table 5]

Embodiment 3 FIG. Production of fermented seasoning K7 strain and MT31 strain were used as yeast. Moromi was prepared as shown in Table 6.

[0029]

[Table 6]

Fermentation control: The fermentation temperature was kept at 15 ° C. throughout. On the 3rd day after the primary preparation, the secondary preparation is carried out.
Three days later, a non-drinking treatment was performed by adding 65 g of sodium chloride.
After another two days, the insoluble components were removed by squeezing filtration,
A fermented seasoning was obtained. Table 7 shows the components of the manufactured fermented seasonings.

[0031]

[Table 7]

Embodiment 4 FIG. Production of bread The K7 strain and the MT31 strain were used as yeast. Culture was performed using a 30-liter jar fermenter according to the following method. Seed culture YPD medium, 30 ° C., shaking, 24 hours Main culture Molasses 3.5 liters (sugar 30%) Urea 20 g KH ₂ PO ₄ 5 g Exponential fed-batch culture 10 hours pH 4.5 Adjusted with ammonia Culture temperature 30 ° C., aeration rate After stirring at a rate of 30 liters / minute 360 rpm, the cells were washed with water and dehydrated to obtain pressed yeast.

Using the obtained yeast, bread was produced in accordance with the following composition and steps. (Blending) Strong flour 100 (parts by weight) Yeast 3 Yeast food 0.1 Sugar 5 Salt 1.8 Skim milk powder 2 Shortening 4 Water 68 (Process) Dough kneading Low speed 3 minutes, medium speed 6 minutes, medium speed 5 minutes ( SC151 type, manufactured by Kanto Mixing Machine Industry Co., Ltd. Kneading temperature 27 ° C Floor time 28 ° C, 40 minutes Bench time Room temperature 15 minutes Wheeler 40 ° C, humidity 85%, 50 minutes Baking 215 ° C, 30 minutes Bread aroma component Put 5 g of crushed bread crumbs into a 20 ml vial and seal
After heating at 0 ° C. for 60 minutes, quantification was performed by head space gas chromatography.

[0034]

[Table 8]

Table 8 shows the results. By using the MT31 strain, bread with higher aroma can be obtained as compared with the case of using the K7 strain.

[0036]

Industrial Applicability According to the present invention, a methyl-threonine-resistant strain belonging to the genus Saccharomyces is used to produce a highly fragrant food or drink containing a larger amount of alcohols (active amyl alcohol, n-propyl alcohol, etc.) useful as flavor components. can do.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI // (C12N 1/16 (C12N 1/16 C12R 1: 865) C12R 1: 865) (58) Field surveyed (Int.Cl. . ^7, DB name) C12G 1/00 - 3/14 C12C 1/00 - 13/10 C12N 1/16 - 1/19 A21D 8/04 A23L 1/23 A23L 1/238 - 1/238 111 A23L 1 / 202 A23L 1/00-1/035 BIOSIS (DIALOG) WPI (DIALOG) JICST (JOIS)

Claims (6)

(57) [Claims] 1. A belonging to the genus Saccharomyces, have a methyl threonine resistance, and n- propyl alcohol and
A method for producing a food or drink, comprising using yeast having an improved ability to produce active amyl alcohol . 2. The method according to claim 1, wherein the food or drink is a food or drink selected from alcoholic beverages, fermented seasonings and bread. 3. The method according to claim 1, wherein the yeast is Saccharomyces cerevisiae.
The method for producing a food or drink according to claim 1, which is a yeast belonging to 4. A belongs to the Saccharomyces Serebije
Yeast is a Saccharomyces Serebije MT31 strain (FERM BP-4364), the preparation of food products of claim 3, wherein. 5. A member of Saccharomyces cerevisiae,
Have a methyl threonine resistance, and n- propyl alcohol
Improved ability to produce alcohol and active amyl alcohol
Yeast . 6. Saccharomyces cerevisiae MT3
1 strain (FERM BP-4364).