WO1998029564A1

WO1998029564A1 - Method for judging early flocculation

Info

Publication number: WO1998029564A1
Application number: PCT/JP1997/004654
Authority: WO
Inventors: Takashi Nakamura; Kazuhiro Chiba
Original assignee: Kirin Brewery Co., Ltd.
Priority date: 1996-12-27
Filing date: 1997-12-17
Publication date: 1998-07-09
Also published as: JPH10179190A; JP3121552B2; AU7891598A

Abstract

A method by which the ability of untreated barley to cause early flocculation, i.e., whether or not a factor causing the early flocculation of yeast is present in untreated barley, can be judged much more easily in a shortened period as compared with the methods of the prior art. Specifically, a method for judging the ability of untreated barley to cause early flocculation by the fermentation test using yeast, which comprises adding an enzyme to the barley to conduct the enzymatic treatment of the barley, adding the product of the enzymatic treatment or a high-molecular fraction separated from the product to a synthetic wort to prepare a sample for the fermentation test, subjecting the sample to the fermentation test, and determining the turbidity of the sample after 48 hours for judging whether or not such a factor is present in the barley.

Description

Description Quick setting method Technical field

The present invention relates to a method for judging the fast setting property of raw wheat used for brewing alcoholic beverages such as beer or whiskey, that is, a method for determining the presence or absence of a fast setting factor in raw wheat. Background art

Alcoholic beverages that are produced by fermentation with yeast using barley malt, such as barley and wheat as a fermentation raw material, are classified as “Happoshu,” which has a lower malt ratio than beer and is classified under “Happoshu” by the Liquor Tax Law. Including whiskey).

When producing such alcoholic beverages using malt as a raw material, a phenomenon called “early flocculation phenomenon” was sometimes observed during the fermentation process. This refers to the phenomenon in which the yeast flocculates and settles in the fermentation process, especially in the late stage of fermentation, even though the sugar that can be assimilated by the yeast still remains in the wort. The sedimentation stops the progress of the fermentation. If this phenomenon is observed, it has been known that fermentation is not sufficient, resulting in non-standard liquor products, and that brewing of beer or the like using wheat with fast-setting properties as raw material has been known to cause severe damage. Many studies have been conducted for a long time to solve the problem of early flocculation phenomenon in the production of alcoholic beverages using malt as a raw material.As a result, this early flocculation phenomenon is derived from raw wheat and contained in malt It was also found that high acid polysaccharides were the cause, but factors that cause the early flocculation phenomenon (hereinafter referred to as “coagulation factors”) mosquito, whether they are present in the raw wheat, or in the malting process It was not known if it would form during. At present, little research has been conducted on a method to solve the early aggregation phenomenon by eliminating the precoagulation factor.

When barley is used as a raw material, it has been conventional to check for the presence of a precoagulation factor in barley and to select only barley malt that does not cause early flocculation. The conventional method of confirming the presence of a precoagulation factor in barley is to actually produce barley on a small scale, prepare wort from the malt, and actually ferment the wort using yeast. The purpose of this study was to confirm the presence of a precoagulation factor in barley from the progress of the barley.

The conventional method described above uses a small scale of actual brewing to check for the presence of a precoagulation factor in barley.Although the reliability of the results is satisfactory, the production of malting and wort It takes about 7 days to confirm the presence of the fast-coagulation factor in barley from the progress of fermentation, and about 8 days to confirm the presence of the fast-coagulation factor in barley. I was

In addition, it is necessary to consider barley dormancy, water sensitivity, etc. in order to produce barley.Because it is impossible to use barley immediately after harvesting for barley production, the above-mentioned conventional method must be applied to barley immediately after harvesting. It is not possible to apply this method to barley.At about 2 months after harvesting, it becomes necessary to check the presence of the pre-coagulation factor in the barley for about half a month. It takes at least about two and a half months to check for the presence of premature coagulation factors in barley.Especially in the case of barley immediately after harvesting, the presence or absence of premature coagulation cannot be determined early. When the barley with curdiness was purchased, the damage was very large.

Disclosure of the invention

The object of the present invention is to achieve a much simpler and shorter Another object of the present invention is to provide a method for judging the fast-coagulation property of raw wheat, that is, the presence or absence of a fast-coagulation factor in the raw wheat.

In the course of research on the early flocculation phenomenon, the present inventors have found that the early flocculation factor that causes the early flocculation phenomenon is not generated in the malting process but is originally present in the raw wheat. That is, when a specific enzyme is added to raw wheat and subjected to an enzyme treatment, the precoagulation factor is extracted without being decomposed, and a fermentation test is performed using the extracted enzyme-treated product. By doing so, they found that it was possible to determine the fast-setting property, and completed the present invention. That is, the present invention relates to a method for determining the fast-coagulation property of raw barley such as barley by a fermentation test using yeast. And an enzyme having at least a combination of a protease and a raw wheat is enzymatically treated, and the resulting enzyme-treated product is used as a part or all of a fermentation test raw material. .

Further, the present invention provides a method for determining the fast-coagulability of raw barley such as barley by a fermentation test using yeast, wherein an enzyme, preferably α-amylase, / 3-amylase, β-glucanase, The raw wheat is enzymatically treated by adding an enzyme having at least a combination of proteases, the polymer fraction of the resulting enzyme-treated product is separated, and the polymer fraction is used as a part of the fermentation test raw material. The present invention relates to a method for judging the fast setting of raw wheat.

Furthermore, the present invention relates to a method for determining the fast setting of raw barley such as barley by a fermentation test using yeast, wherein an enzyme, desirably monoamylase, / 3-amylase, β-glucanase and protea is added to the raw barley. A raw wheat is enzymatically treated by adding an enzyme having at least a combination of enzymes, and the resulting enzyme-treated product or a polymer fraction separated from the enzyme-treated product is added to synthetic wort to conduct a fermentation test. The present invention relates to a method for judging the fast-coagulation property of raw wheat, which comprises determining the presence or absence of a fast-coagulation factor in the raw wheat by measuring the turbidity of the raw material for the fermentation test 48 hours later.

In the present invention, the raw wheat such as barley and wheat is not limited to the one that has been harvested for a certain period after harvesting, and can be used immediately after the harvest. In addition, the amount of raw wheat required for judging fast-coagulation may be about 20 g, but it is better to prepare about 50 g to further increase the precision of the experiment.

It is desirable that the raw wheat is first ground to such an extent that the enzyme treatment can be carried out efficiently, specifically, 90% by weight or more passes through a sieve having a pore size of 0.547 mm. Next, the raw material barley is suspended in water, preferably in warm water at about 40 to 60 ° C., and an enzyme (agent) is added thereto to carry out an enzyme treatment.

In the present invention, as the enzyme to be added, any enzyme can be used as long as it has an action of extracting the fast-coagulation factor from the raw wheat without being decomposed. It is desirable to use one containing at least a combination of amylase, / 3-glucanase and protease.

The enzyme to be added is not limited to a high-purity enzyme as long as it has enzyme activity.For example, an enzyme-containing treated cell may be used, or a mixture of enzymes having different actions may be used. However, it is a condition that the raw wheat crushed material is not gelatinized in water without decomposing the early simulation factor.

The enzyme to be used will be described in more detail. As an enzyme containing α-amylase, protease and —glucanase, Ceremix 6 XMG (manufactured by Novo) is used. As glucanase, A product derived from Noregillus. A spergi 1 lusniger (Finizym, manufactured by Novo) is a β-Rolase derived from barley (bar 1 ey). Company) can be exemplified.

The enzyme treatment conditions such as the amount of the enzyme added, the reaction temperature, and the reaction time vary depending on the properties of the enzyme to be used, but it is preferable to carry out the reaction under conditions where the enzyme activity can be sufficiently exhibited. For example, when the above enzyme is used, it is sufficient to react at about 40 to 60 ° C for about 2 to 3 hours or more, but the temperature is changed stepwise to simulate a normal saccharification process. Of course it is possible.

After filtering the enzyme-treated product obtained by the enzyme treatment using a filter paper or the like, the enzyme activity is deactivated by heating such as boiling, and the mixture is filtered again to remove the heat coagulated product by heating. If the raw wheat has precoagulation properties, the high-coagulation factor is contained in the high-molecular fraction in this filtrate (hereinafter referred to as “pseudo wort”). After mixing with the fermentation raw material, a normal fermentation test using yeast can be performed to determine the presence or absence of a fast-coagulation factor in the raw wheat.

In addition, after separating only the high-molecular fraction from the above-mentioned pseudo-wort, it is added to another fermentation raw material as a part of the fermentation test raw material and subjected to a fermentation test, whereby the presence or absence of the fast-coagulation factor is more accurately determined. Can be determined. As a specific method for separating the high-molecular fraction, about twice the volume of ethanol is added to the above-mentioned pseudo-wort, and the high-molecular fraction is stirred for about 5 minutes. Examples of the method of obtaining the polymer fraction from the pseudo wort include the method of obtaining the polymer fraction as a precipitate. In addition to the above-mentioned precipitation with ethanol, the polymer fraction such as dialysis and ultrafiltration may be used. Any method that can be separated can be used. If the raw wheat has fast-coagulating properties, this polymer fraction contains a fast-coagulating factor. Will be rare.

As a medium to which the high-molecular fraction for the fermentation test is added, ordinary wort is of course not a problem, but in order to make the results reproducible and accurate, other than malt must be used. Synthetic wort (eg, We infurtner, F., et. A1-. Brauw issenschaft, 14, 109 (1961)) prepared with ingredients (sugars, amino acids, inorganic salts, etc.) It is desirable to use it. A method for preparing synthetic wort obtained by partially improving synthetic wort such as Weinfurtner is described below.

(Preparation method of synthetic wort)

Prepare in advance the following solutions A, B, C, and D, respectively. Solution C and solution D should be made into a stock solution, stored in a refrigerator, and measured in a clean bench.

Next, each prepared solution A: 700 m and solution B: 50 m], solution C: 5 ml and solution D: 1001 were mixed, adjusted to 800 ml with distilled water, and adjusted to pH 5. Adjust to 7.

A liquid:

D (-) — Fructose 2 g

D (+) — glucose 8 g

4 g of sucrose

Malt monohydrate 6 4 g

Dextrin 2 7 g

3.5 g of casamino acid

4 g of peptone

C a C 1 ₂ (anhydrous) 1 g

KC 1 1 g Dissolve in distilled water to adjust the volume to 700 ml, and sterilize in an autoclave. B liquid:

Mg S 0 ₄ 7 H ₂ 0 1 g

Dissolve in distilled water, adjust to 50 ml, and sterilize in an autoclave. C liquid:

Wild boar 500 mg

(+) — Calcium pantothenate 500 mg

Nicotinic acid 50 mg

Thiamine hydrochloride 50 mg

Pyridoxine hydrochloride 50 mg

(+) —Piotin 50 mg

Peracil 25 mg

Guanin 25 mg

Dissolve in distilled water to adjust the volume to 250 ml, and sterilize the filter.

D solution:

H ₃ BO 100 mg

Z n SO, 7 H ₂ 0 1 00 mg

Mn C 1-4 H ₂ 0 100 mg

F e C 1 50 mg

_{C u S 0 4 · 5 H} 2 〇 1 0 mg

K I 10 mg

Is dissolved in distilled water to adjust to 1000 ml, and then sterilized in a autoclave. <As for the determination method after the fermentation test, a conventionally known method (for example, Kori mo to, et. Rept.Res.Lab.Kirin Brewer Co., ltd., 18, 63 (1975)) Can be used. As such conventional methods, the above-mentioned pseudo-wort is directly used as a fermentation test raw material (medium), or the above-mentioned pseudo-wort or the above-mentioned high-molecular fraction is added to wort obtained from raw wheat having no fast-setting property. For fermentation test

(Medium), fermented at about 8 ° C for 8 days using yeast, and comprehensively determine the growth rate of yeast by measuring turbidity and sugar content in the raw material (medium) Is mentioned.

In the fermentation test using yeast, especially when using the high-molecular fraction, it was confirmed that accurate determination can be made by measuring the turbidity after fermentation at a temperature of 20 ° C for 48 hours. In this case, use a turbidimeter to measure the absorbance at a wavelength of 800 m.

(OD 800) is measured, and a value obtained by subtracting the OD 800 of the test plot from the 〇D 800 of the control plot is compared with D PF (Degre e o f P r e m a t u r e F l o c c u l a t i o n) 48. At this time, as a comparative example, it is desirable to simultaneously test normal barley or early-cured barley to compare the turbidity. BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited to such examples. Example

A commercial enzyme containing β-glucanase (FFnizym; manufactured by Novo) and a protease, α-amylase, and / 3-glucanase are contained in 300 ml hot water heated to 55 ° C. 0.5 g of each of the commercially available enzymes (Cerixix 6 X MG; manufactured by Novo) and 500 units of / 3-amylase (/ 3-amylase; manufactured by Sigma) were added. To this, add 50 g of barley finely ground by a disk mill, stir well to make it uniform, and mix at 55 ° C for 3 hours. The enzyme treatment was completed to obtain an enzyme-treated product.

The enzyme-treated product was stirred well and filtered using filter paper (Toyo Filter Paper No. 2), and 180 ml of the filtrate was accurately collected. The 180 ml filtrate collected was heated and boiled until the liquid volume was reduced to less than half, then the total volume was adjusted to 100 ml and filtered again in the same manner to make pseudo wort. I got

While stirring the pseudo wort, 200 ml of ethanol was added little by little, and after stirring for 5 minutes, centrifuged, the supernatant was discarded, and 10 ml of boiling water was added to the precipitate. After dissolving the precipitate and adjusting the total volume to 25 m1, centrifugation was performed again, and the supernatant (hereinafter, referred to as "barley high-molecular-weight fraction extract") was subjected to the fermentation test.

To 80 ml of the synthetic wort prepared by the above-described method, 20 ml of the above-mentioned barley polymer fraction extract was added to adjust the pH to 5.7, and this was used as a test plot. On the other hand, distilled water was used in place of the barley polymer fraction extract and the pH was adjusted to 5.7 to prepare a control. After adding 0.35 g of brewer's yeast to these, they were put into a fermentation tube (100 m1 volume) with a diameter of 27 m1 and fermented at 20 ° C. After 48 hours, 5 cm from the liquid level OD 800 was measured by taking 2 ml each from the place. The value of the control plot was subtracted from the value of the test plot to obtain a value DPF48 indicating the degree of fast-coagulation.

Seven types of barley, which were checked for the presence of premature setting by on-site malting, were used as samples and tested according to the above, and the correlation with the actual premature setting was examined. Table 1 shows the results. As can be seen from Table 1, there is a high correlation between the value of DPF 48 and the actual fast setting, and it was confirmed that barley having a fast setting factor was found when the DPF 48 value was large. . It has also been confirmed that the reproducibility of these results is very high. 【table 1】

Industrial applicability

According to the present invention, it has become possible to accurately and reproducibly determine the fast-coagulation property of raw wheat in about 5 days, that is, the phenomenon of yeast agglomeration in the fermentation of alcoholic beverages using wheat as a raw material. Further, according to the present invention, it is possible to determine the quick setting of raw wheat just after harvesting or even a small amount of raw wheat.

Claims

The scope of the claims

1. In a method for determining the fast-coagulation property of raw wheat by a fermentation test using yeast, an enzyme is added to raw wheat and the raw wheat is subjected to enzymatic treatment. A method for judging the fast-setting properties of raw wheat, which is characterized in that:

2. In a method of determining the fast-coagulation property of raw wheat by a fermentation test using yeast, an enzyme is added to raw wheat and the raw wheat is subjected to enzymatic treatment, and the polymer fraction of the resulting enzymatically treated product is separated. A method for judging the fast setting of raw wheat, wherein the high molecular fraction is used as a part of the raw material for fermentation test.

3. In a method for determining the fast-coagulation property of raw wheat by a fermentation test using yeast, an enzyme is added to raw wheat and the raw wheat is subjected to enzymatic treatment, and the resulting enzymatically treated product or separated from the enzymatically treated product is obtained. The high-molecular-weight fraction is added to synthetic wort as a fermentation test raw material, and the presence or absence of a fast-coagulation factor in the raw wheat is determined by measuring the turbidity of the fermentation test raw material 48 hours later. Method for determining the fast setting of raw wheat.

4. The raw wheat as claimed in any one of claims 1 to 3, wherein the enzymatic treatment is carried out using an enzyme having at least a combination of α-amylase, / 3-amylase, / 3-glucanase and protease. Coagulation determination method.

5. The method for judging the fast setting of raw wheat according to any one of claims 1 to 4, wherein the raw wheat is barley.