JPH0889230A - Production of sake - Google Patents

Abstract

PURPOSE: To obtain SAKE(Japanese rice wine) containing a large amount of a flavoring component such as ferulic acid, its ethyl ester, vanillin or vanillic acid and having a mellow flavor by reacting a hydroxycinnamic ester hydrolase in a step for saccharifying, fermenting and aging unrefined SAKE for SAKE. CONSTITUTION: A hydroxycinnamic ester hydrolase in an amount of >=1 unit, preferably 3-100 units based on 1kg raw material rice is added and reacted in a step for saccharifying, fermenting and aging an unrefined SAKE for SAKE. Thereby, SAKE containing a relatively large amount of a flavoring ingredient having depth is obtained without using a specific yeast.

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 sake, and more particularly to a method for producing sake with rich flavor components such as ferulic acid, ethyl ferulic acid, vanillin and vanillic acid.

[0002]

2. Description of the Related Art Conventionally, in order to improve the flavor of sake, isoamyl acetate having a high aromaticity has been used by using a novel yeast having high aromaticity bred by crossing haploid strains derived from sake yeast 701 and sake yeast 901. And a method of obtaining sake having a high aromatic content and containing a lot of ethyl caproate are known (Japanese Patent Laid-Open No. 5-31).
No. 7034). Also, isoamyl alcohol,
A method for producing sake characterized by using a mutant yeast which produces a large amount of a flavor component such as isoamyl acetate (Japanese Patent Laid-Open No. Sho 62-62).
No. 6669) is known.

[0003]

However, these methods have the drawback that they require sophisticated techniques to create specialized yeast.

Therefore, an object of the present invention is to obtain sake having a rich flavor without creating a special yeast.

[0005]

Means for Solving the Problems As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that the action of adding hydroxycinamic acid ester hydrolase in the process of normal saccharification and fermentation of sake. Based on this finding, the present invention was completed based on the knowledge that when simmering, fermented acid, ferulic acid ethyl ester, vanillin, vanillic acid, and the like can be obtained. That is, the present invention is a method for producing sake, which comprises adding a hydroxycinamic acid ester hydrolase in the process of saccharification, fermentation and aging of sake.

The present invention will be described in detail below. First, as the sake liquor referred to in the present invention, normal polished rice is washed, soaked to swell, and then steamed by a conventional method to obtain steamed rice,
Part of this is koji, and part is rice with water,
An acid and sake yeast are added to this, and the yeast is selectively cultivated with a sugar solution containing acid (to be originally fermented) to prepare an aged sake mother. One example is the mash that has been able to prepare three people for water in three stages (each called Hatsuzo, Nakazoe, and Tomezo). It should be noted that the mash made from koji, rice, and water can be divided into four stages for the sake mother.

The mash thus obtained is subjected to saccharification with koji and liquor fermentation with yeast in parallel by an ordinary method, and then subjected to aging, squeezing, filtering, burning, etc. to obtain sake. .

In order to carry out the present invention, a culture of a microorganism of the genus Aspergillus or a processed product thereof having the ability to produce a hydroxycinamic acid ester hydrolase in the above-mentioned process of saccharification, fermentation and aging of sake (in the present invention, these are Collectively referred to as hydroxycinnamic acid ester hydrolase).

The hydroxycinamic acid ester hydrolase used in the present invention has already been disclosed by the present inventors in JP-A-57-
No. 99193, which is disclosed in detail, for example, Aspergillus japonicas ATCC20236,
A culture obtained by subjecting strains such as Aspergillus niger-IAM3002 and Aspergillus cytoii ATCC14332 to solid culture or liquid culture according to a conventional method using a nutrient medium used for culture of filamentous fungi is used as it is.

Alternatively, celite or the like may be added to the water extract of the solid culture or the filtrate of the liquid culture and clarified by filtration to obtain a crude enzyme solution. Ammonium sulfate or the like may be added to the water extract or the crude enzyme solution. A crude enzyme powder obtained by (salting out method) or adding alcohol (organic solvent precipitation method) to obtain a precipitate, which is collected by vacuum drying and the like is used.

Alternatively, after the crude enzyme powder is dissolved in a buffer solution or the like, an ion exchange Sephadex such as DEAE-Sephadex A-50, QAE-Sephadex A-50 or CM-Sephadex, or Sephadex G- is used. Two
Individual means such as gel filtration using Sephadex such as 00, isoelectric focusing using ampholine, electrophoresis using acetate membrane starch, acrylamide gel, and other isoelectric precipitation, or A purified enzyme is obtained by appropriately utilizing these appropriate combinations and is used (in the present invention, the above-mentioned crude enzyme and purified enzyme are referred to as "treated product of culture").

[0012] These microorganisms have a hydroxycinnamic acid ester hydrolase activity in the koji of 0.
When using a microorganism having a hydroxycinamic acid ester hydrolase-producing ability of 5 units / g or more of koji, a large amount of the enzyme can be produced and accumulated in the culture, so that it may be used as a crude enzyme or a purified enzyme. It is preferable because it does not need to be in such a form and can be added and act as it is.

A Method for making koji: 36 g of 80% sprinkled bran was put in a 1 liter Fernbach flask, sterilized under pressure at 120 ° C. for 1 hour, and after cooling, inoculated with Aspergillus microorganisms, and 72 at 30 ° C. Incubate for hours.

B Hydroxycinamic acid ester hydrolase activity assay method Agricultural biological chemistry-46
Vol., P. 297 (1982). Commercially available chlorogenic acid (3-caphoeoylquinic acid) (manufactured by Tokyo Kasei Kogyo Co., Ltd.) was dissolved in a 0.05 M phosphate buffer (pH 6.5) at a concentration of 530 mg / l. 0.03 ml of enzyme solution dissolved in the same buffer solution in 0.5 ml of this solution
Is added and reacted at 30 ° C. After a certain period of time, 10 ml of 80% methanol water was added to stop the reaction, and the absorbance at 350 nm was measured. In the above method, the absorbance was measured in exactly the same manner except that the "enzyme solution was heat-inactivated by holding the enzyme solution at 70 ° C for minutes" was used in place of the "enzyme solution" to give a blank value. The activity is defined as 1 unit (U) of the enzyme capable of hydrolyzing 1 micromol of the substrate in 1 minute. The enzyme activity is calculated by the following formula. t2-t1: Reaction time (min) Et1: Value obtained by subtracting blank value from OD value at t1 time Et2: Value obtained by subtracting blank value from OD value at t2 time

The above-mentioned enzyme to be added and acted in the sake saccharification and fermentation aging step varies depending on the form, the specific activity of the enzyme, the method of making the enzyme act, etc., but it is preferably 1 unit or more per 1 kg of the starting rice rice. Is 3 to 10
It is preferred to use 0 units.

The time of addition may be any time during the saccharification and fermentation ripening step, but if added during the initial stage of preparation, that is, during the initial addition, Nakazo or distillation, free ferulic acid derived from the raw material is added. It is preferable because a large amount can be produced and accumulated in the mash.

[0017]

[Function] When a hydroxycinamic acid ester hydrolase is added and used in the sake saccharification and fermentation ripening process as described above, the enzyme acts on the raw material rice to produce and accumulate free ferulic acid derived from the raw material in the mash. Excuse me. And, it is known that this ferulic acid itself has a sweet fragrance, but part of the ferulic acid is decomposed into vanillic acid, a substance having a sweet flavor such as vanillin, so that the flavor of sake is gradually improved. On the other hand, ferulic acid and vanillic acid react with the alcohol present in sake to form ferulic acid ethyl ester and vanillic acid ethyl ester, each of which has a heavy flavor.

Therefore, according to the present invention, it is possible to obtain sake having a heavy sweet scent, that is, a rich flavor.

Hereinafter, the present invention will be described more specifically by showing examples. The cultures of Aspergillus microorganisms having a high productivity of the hydroxycinamic acid ester hydrolase used in the following examples, and the crude enzyme powder of the enzyme were prepared by the following method.

(Aspergillus japonicas AT of Aspergillus niger
Preparation of culture of CC20236 strain) 120% sprinkled bran in a sterilized flat plastic container
After sterilizing under pressure for 1 hour at ℃, put what was cooled, inoculate the strain of Aspergillus japonicas ATCC20236 into this, and cultivate for 72 hours at 30 ℃ while stirring occasionally, hydroxycinamic acid ester hydrolase activity, Two
A solid culture showing units / g koji was obtained.

(Preparation of Crude Enzyme Powder of Hydroxycinamic Acid Ester Hydrolase) 5 times by weight of water was added to the above solid culture and left for 2 hours at room temperature with occasional stirring to obtain juice. Celite is added to the filtrate and filtered to obtain a transparent enzyme solution. The enzyme solution was cooled to 5 ° C., and 3 volumes of alcohol cooled to the same temperature was added thereto, and the produced precipitates were collected and vacuum dried at a low temperature of 20 ° C. to obtain a crude enzyme powder.

[0022]

Example 1 (Manufacturing Method of Sake with Rich Flavor) (1) Preparation of Rice Koji 225 g of polished white rice was washed, dipped, drained, steamed and allowed to cool, and then commercially available. The rice koji was obtained by inoculating the seed koji mold for sake and making the koji by a conventional method. (2) Preparation of steamed rice 450 g of whitened steamed rice was washed, soaked, drained, steamed and allowed to cool by ordinary methods to obtain steamed rice. (3) Preparation of liquor Mother 700 ml of pumped water was mixed with the above rice koji and the above steamed rice, and 20 ml and 4 ml of seed yeast (2 × 10 ⁸ cells / ml) and lactic acid (10-fold diluted solution) were mixed therein. , 10-1
Saccharification and fermentation was carried out at 5 ° C. for 15 days, and the yeast was selectively grown in a sugar solution containing an acid to prepare a liquor. (4) Manufacture of sake (first addition) To 1200 ml of the above-mentioned sake mother, 400 g of rice koji (raw rice equivalent),
Mix 950 g of steamed rice (same) and 1150 ml of pumped water,
This was fermented for 48 hours. (5) Same (Nakazoe) 2075 g of steamed rice (converted to raw rice), rice malt 625 in addition to Hatsuzou
g (the same) and 3250 ml of pumped water were mixed, and this was added to 24
Fermented for hours. (6) Same (Tozoe) In addition to Nakazoe, steamed rice 3450 g (raw rice equivalent), rice koji 82
5 g (same) and 6250 ml of pumped water are mixed, and this is 1
The mixture was fermented and aged at 0-15 ° C. for 20 days and squeezed and filtered by a conventional method to obtain a beer juice (sake). (7) When preparing the above-mentioned aged mash, at the time of distillation, 50 units of crude enzyme powder of hydroxycinamic acid ester hydrolase was added to the present invention group 1, 100 units of the same enzyme to the present invention group 2, and the control group. The crude enzyme powder was not added to the above, but allowed to stand. (8) A soup was sampled from the thus-obtained aged mash, the alcohol and aroma components thereof were analyzed, and a sensory test was conducted. (9) The alcohol in the liquid broth was analyzed by the National Tax Agency analysis method, and the aroma components ferulic acid, vanillin, and vanillic acid were treated with O. D. S was measured using high performance liquid chromatography using a resin for separation (TSKgel ODS-120T manufactured by Tosoh Corporation). (10) In addition, a sensory test is performed by a panel of 10 persons having discrimination ability, and the flavor of sake is evaluated by a scoring method.
Compared to the control group, +2 is excellent, +1 is slightly excellent, 0 is no difference, -1 is slightly inferior, -1 is inferior.
It was evaluated as 2, and the average value was shown. The results are summarized in Table 1.

Ingredients Sake Mother Hatsuzou Nakazoe Tosekizo Total Total Rice 675g 1350g 2700g 4275g 9000g Kakemi 450g 950g 2075g 3450g 6925g Koji Rice 225g 400ml 625ml 825g 2075g 1 50ml 150ml 6 50ml 150ml 6

[0024]

[Table 1]

From the results of Example 1, when a hydroxycinnamic acid ester hydrolase was added and used during the saccharification and fermentation aging of sake, a heavy flavor component of ferulic acid ethyl ester, vanillin and vanillic acid was added to the beer juice. It can be seen that the sake is enriched and the sake with good flavor can be obtained.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Yoshihiko Endo, 339 Noda, Noda, Chiba Prefecture, Kikkoman Corporation (72) Yoshimi Shimazu, 339, Noda, Noda, Chiba Prefecture, Kikkoman Corporation

Claims (2)

[Claims] 1. A method for producing sake, which comprises adding a hydroxycinamic acid ester hydrolase in the step of saccharification, fermentation and aging of sake. 2. The method for producing sake according to claim 1, wherein 1 unit or more of hydroxycinamic acid ester hydrolase is added to 1 kg of the raw material rice.