JPS63188378A - Production of must wine - Google Patents

Abstract

PURPOSE:To obtain must wine free from peculiar smell of must, by fermenting must by the use of gel having immobilized both alpha-acetolactate decarboxylase and a yeast cell. CONSTITUTION:Must, concentrated wine juice to be a raw material for wine brewing, is fermented by using gel having immobilized both alpha-acetolactate decarboxylase and a yeast cell. Various kinds of gel such as potassium alginate, K-carrageenan, etc., can be used as the gel. Any yeast can be used as the yeast so long as the yeast is yeast belonging to the genus Saccharomyces cerevisiae, etc. alpha-Acetolactate decarboxylase produced by a bacterium belonging to the genus Corynebacterium, etc., may be used as the alpha-acetolactate decarboxylase. The fermentation is carried out by diluting must into 10-35 deg. Brix and pH 2.8-3.8 and bringing the gel having immobilized both the alpha-acetolactate decarboxylase and the yeast cell into contact with the must. Then after the fermentation is over, the fermented solution is filtered to give must wine.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to a method for producing must wine using must as a raw material. Must is concentrated grape juice that is a raw material for winemaking and contains 1% (V/V) or more of alcohol. Must wine produced using this must as a raw material has an unpleasant aroma called must-specific odor. The present invention provides a method for producing must wine that does not have a must-specific odor.

Conventional technology The causative agent of must wine's unique odor is unknown, but it is known to increase when must wine is heated, to be produced after the middle stage of must fermentation, and to be present in wine made from raw grape juice. has been shown to be undetectable. It has also been revealed that precursors or production-promoting substances of substances that cause the unique must odor are formed during the storage of fruit juice under high concentration of sulfur dioxide before must production (concentration) (1985 Fermentation Engineering Society Collection of conference lecture abstracts, page 38).

As a method for producing must wine, conventional methods using electrodialysis (Japanese Patent Laid-Open No. 55-77
884), a method using a large amount of sake mash (Japanese Patent Application Laid-Open No. 58-15
5074), a method of fermenting by adding yeast extract (
JP-A-58-155075), a method of fermentation under continuous or intermittent reduced pressure (JP-A-58-155076, JP-A-58-165783), a method of using yeast with strong alcohol tolerance as a yeast mother (Japanese Patent Application JP-A-61221423) ,
A method of fermentation by adding valine, isoleucine, fresh grape juice or its fractions (Patent Application No. 1-15
8535) etc. are shown.

Furthermore, it is known that during the fermentation of alcoholic substances such as wine and beer, diacetyl is often produced in excess of a threshold value. This is produced non-enzymatically from α-acetolactate formed by wine yeast, beer yeast, etc. Diacetyl has a strong and unpleasant odor and therefore has an undesirable effect on the aroma and taste of products.

α-acetolactate decarboxylase (EC4,1
, 1, 5) is an enzyme that decarboxylates α-acetolactate and generates acetoin, so it can directly convert α-acetolactate to acetoin and reduce the production of diacetyl. A method for producing beer and wine with low diacetyl content is known that takes advantage of this property (Tokukaiho 5
7-501114).

Problems to be solved by the invention Since the amount of fresh grape juice obtained in Japan is limited, and the harvest period is also limited, must wine made from must that is available in large quantities regardless of the season. Various manufacturing methods have been developed.

However, must wine has the drawback of being accompanied by an unpleasant odor called must odor. Therefore, it is desired to develop a method for producing must wine that does not have a must odor.

Means for Solving the Problems As a result of research into a method for producing must wine that does not have a must-specific odor, the inventor discovered that fermentation using a gel in which α-acetolactate decarboxylase and yeast cells were co-immobilized was conducted. The present invention was completed based on the discovery that it is possible to produce must wine that does not have a must odor by carrying out the following steps.

The present invention will be explained in detail below.

The present invention provides a method for producing must wine by fermenting must using a gel co-immobilized with α-acetolactate decarboxylase and yeast cells.

No effect will be observed unless α-acetolactate decarboxylase and yeast cells are co-immobilized in the gel. A method in which must is fermented with a gel on which yeast cells are immobilized and then brought into contact with a gel on which α-acetolactate decarboxylase is immobilized; It is not possible to produce must wine that does not have a must-specific odor by any of the fermentation methods using a gel that has immobilized must wine. Moreover, even if must wine produced by a normal brewing method is brought into contact with a gel in which α-acetolactate decarboxylase is immobilized, the must-specific odor is not reduced. Furthermore, α-acetolactate decarboxylase and yeast cells are allowed to coexist without immobilization,
Even if must is fermented, must wine that does not have a must-specific odor cannot be obtained.

Gels for co-immobilizing α-acetolactate decarboxylase and yeast cells include calcium alginate, K-
Various gels such as carrageenan and collagen can be used.

As a yeast, Satucharomyces cerevisiae or Satucharomyces criveeri (The Ys)
Yeasts), Craigherv Henry, El Sepia
According to the classification of Science Publishers (1984)], any yeast can be used.

α-acetolactate decarboxylase is produced by microorganisms belonging to the genus Corynebacterium, Brevibacterium, Lactobacillus, etc.
European Brewery Conventi
onproceeding of 19th Cong
ress 1983. p, 161, patent application No. 6166
808), etc., but preferably the optimal p
It is desirable that H be as close as possible to the pH of the must (usually about 2.8 to 3.6).

To co-immobilize α-acetolactate decarboxylase and yeast cells, α
- It is carried out by preparing a gel of acetolactate decarboxylase and yeast cells. A co-immobilized gel can be prepared by a commonly used gelling method. For example, an α-acetolactate decarboxylase enzyme solution and a suspension of yeast cells are mixed in a solution containing sodium alginate etc. so that the maximum concentration of sodium alginate is about 2 to 3.5%. It can be obtained by dropping the solution into a solution containing calcium chloride or the like. At this time, the enzyme concentration in the solution is 100 units/
Although an effect is observed if the concentration is 1 unit or more (the amount of enzyme that produces 1 μmo I of acetoin per minute at 37° C. is 1 unit), a concentration of 1000 units/ml or more is desirable.

The concentration of yeast cells in the solution is 1×103 to 1×10I0 cells/m I, preferably 1×106 to 8×107 cells/m
It is 1.

Contact between the co-immobilized gel and the mast is achieved by packing the gel into a column,
Any method can be used, including a method in which the mast is passed continuously from the bottom to the top of the column, a method in which the gel is suspended in the mast, and a method in which the gel is placed in a basket made of stainless steel or the like and submerged in the mast.

Both red and white must can be used for fermentation. For fermentation, mix the must with 1O-La5°Br1x
, by diluting the gel to pH 2.8 to 3.8 and contacting it with the co-immobilized gel described above. After fermentation is complete, must wine can be obtained by filtering the fermentation liquid.

Examples of the present invention are shown below.

Example <Preparation of α-acetolactate decarboxylase enzyme preparation> Brevibacterium acetylicum (Brev iba
cte-rium acetyl icum) A T
CC954, powdered bouillon 2%, zinc sulfate 0.00
Aerated and agitated (1
81/min.

300 rpm).

After culturing, the cells were collected by continuous centrifugation (8.00 rpm, 20 minutes) to obtain about 190 g (wet weight) of cells.

Using glass beads of 0.1 to 0.2 mm,
Dyno Mill (Ill, A, manufactured by Bachofen)
After disrupting the cells, the residue was removed by centrifugation to obtain a cell-free extract. Ammonium sulfate was added to this cell-free extract to reach 80% saturation, and centrifugation was performed to obtain a precipitate. This is 0. OIM)! J Soot-acid buffer (pH 7
,,O) and dialyzed with the same buffer for 2 days.

Ammonium sulfate was added to the internal solution to reach 60% saturation, and the resulting precipitate was discarded by centrifugation.Ammonium sulfate was further added to the supernatant to reach 80% saturation, and centrifugation was performed to obtain a precipitate. . This is the minimum amount of 0.01M)!
DEAE cellulose (Se
The active fraction was collected by passing through a column (manufactured by rVa) and eluting with the same buffer containing 0.3M sodium chloride.

Measurement of the enzymatic activity of α-acetolactate decarboxylase involves measuring acetoin produced by the enzymatic reaction with mjs
Terfeld method [Journal of Biological Chemistry (J, Biol, Chem,) 16
1.

495 (1945):]. The obtained active fraction was centrifuged after adding ammonium sulfate to 80% saturation.
H7,0) was used in the following studies as an α-acetolactate decarboxylase solution (hereinafter sometimes referred to as enzyme solution).

Note that all the series of operations after completion of the culture were performed at 10°C or lower.

Table 1 shows the enzyme activity and protein concentration of the obtained enzyme solution.

Table 1 23.2 4997 <Immobilization of enzyme and yeast cells> Add 63 ml of the enzyme solution obtained above to 60 ml of a solution (pH 7.0) containing 7% sodium alginate in 0.1 M') acid buffer. and Satsucharomyces cerevisiae OC2 strain (
A bacterial solution (purchased from Japan Brewing Association) suspended in 0.1 M IJ acid buffer at 7 x 10 B cells/ml (pH
7,0> 20 ml was added, shaken, and then added dropwise to a 5% calcium chloride solution to prepare a calcium alginate gel in which α-acetolactate glutoxylase and yeast were co-immobilized. Note that all these operations are 10
The temperature was below ℃.

Fermentation of must using co-immobilized gel〉 The entire amount of gel obtained by the above procedure was placed in a stainless steel cylindrical basket, and placed in a column (φ32 x 270 mm).
) was filled. Continuous fermentation was carried out using white must diluted to 19°Br1x at 25°C in ascending mode (SV O, 32).

After the produced must wine was filtered through a 0.45μ membrane filter, analysis and sensory tests were conducted.

Table 2 shows the analysis results, and Table 3 shows the sensory test results. As a control, white must wine produced by a conventional fermentation method was used.

In addition, white must wine was produced by allowing yeast cells and α-acetolactate decarboxylase to coexist on must without co-immobilizing them, and a gel was prepared in which yeast cells and α-acetolactate decarboxylase were each immobilized separately. , White must wine produced by contacting yeast immobilized gel and then contacting α-acetolactate decarboxylase immobilized gel, and contacting yeast immobilized gel after contacting α-acetolactate decarboxylase immobilized gel. The sensory tests for the white must wines produced by contacting them were conducted in the same manner, and the results are shown in Table 3.

By the method of the present invention, must wine containing 9.0% (V/V) or more of alcohol and having no must-specific odor could be produced.

Table 2 Specific gravity 1.009
1.011 Alcohol (V/V%') 9.4
9.1 extract (W/V%>
5.8 6.2 Total acid (W
/V%) 0.546 0.5100D,
,, 0.168 0.117pH 3,133,00 Table 3 According to the present invention, must wine that does not have a must odor can be produced.

Claims (1)

[Claims] A method for producing must wine, which comprises fermenting must using a gel co-immobilized with α-acetolactate decarboxylase and yeast cells.