JPS6321468B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing a new type of mirin containing more non-fermentable oligosaccharides than glucose. More specifically, an enzyme agent containing transglucosidase, α-amylase, and acidic protease can be added as is or by replacing part or all of rice malt with mirin to produce isomaltose, panose, and isomaltose. This is a novel method for producing mirin, which is characterized by containing a large amount of triose, isomaltotetraose, and other non-fermentable oligosaccharides. Conventional mirin has over 80% of its sugar content being glucose, and has the drawback of lacking a rich flavor. The present inventors conducted extensive research aimed at producing zurumirin with a strong body taste while softening the harsh sweetness from glucose by changing the sugar composition. As a result, transglucosidase, α-
By adding an enzyme agent containing amylase and acidic protease as is, or replacing some or all of the rice malt, and preparing mirin,
We succeeded in producing mirin with a low glucose content of 20 to 30% of the sugar content, a refreshing sweetness, and a rich flavor. Enzymes have been used in the production of mirin for some time, but the purpose was to replace rice malt and improve the utilization rate of raw materials.
There has never been an attempt to produce a new type of mirin by actively changing the ingredients of mirin as in the present invention. Therefore, the enzyme agents used are limited to α-amylase, glucoamylase, and acidic protease, and the use of transglucosidase was completely unknown. The main purpose of the present invention is to produce mirin containing a large amount of non-fermentable oligosaccharides. It was found that glucose was produced and relatively decreased. Additionally, since mirin moromi does not involve alcoholic fermentation, the glucose produced will never disappear. Therefore, in order to minimize the production of glucose, the presence of glucoamylase in the mash must be avoided as much as possible. Therefore, there are limits to the amount of rice malt used. Also, α-amylase is necessary to scald steamed rice.
It was also found that when acidic protease is used in combination with this, it promotes the melting of steamed rice and has a significant effect on the production of non-fermentable oligosaccharides. The present invention was completed based on these findings, and by using acidic protease in combination with transglucosidase and α-amylase, we succeeded in improving the raw material utilization rate and increasing the content of non-fermentable oligosaccharides. . In the present invention, transglucosidase,
An enzyme agent containing α-amylase and acidic protease may be added as is, or may be prepared by replacing part or all of the rice malt. The enzyme agent used contains α-, which is necessary for liquefying steamed rice.
It includes amylase, and the transglucosidases and acid proteases required for the production of non-fermentable oligosaccharides. As the α-amylase in the enzyme preparation, any type of α-amylase with a low glucose-producing ability may be used, but from the economic point of view, those derived from plants or microorganisms are preferable. Specifically, α-amylase from malt, filamentous fungi, or Bacterial α-amylase, actinomycete and bacterial β-amylase, etc. may also be included. Transglucosidase is obtained by culturing Aspergillus cytoi IAM2241, a type of Aspergillus aspergillus, in a liquid medium such as corn meal or CSL, but since the crude enzyme contains glucoamylase, it must be purified before use. There is a need. Simply put, the purification method is to pass the culture solution through a basic anion exchange resin such as Diaion WA-20 to remove acidic substances such as organic acids, and then pass it through an acidic cation exchange resin such as Amberlite CG-50. By this, transglucosidase containing almost no target glucoamylase can be obtained. The acidic protease used in the present invention can also be used regardless of its origin, but microbial ones are preferred, and specifically, filamentous fungi such as Aspergillus layer, Rhizopus layer, Mucor layer, Penicillium genus, etc. acidic proteases. In the present invention, α-amylase, transglucosidase, and acidic protease may be added individually, or may be used in combination. What is convenient for use is a mixture of each enzyme unit in an optimal combination. The amount of each enzyme added may be in excess, but is optimally expressed in units. α-amylase is 100% for 1g of total rice.
~300 units, transglucosidase is also 100
~300 units, acid protease 200-600 units. In normal mirin preparation, these enzymes are added to the alcohol or shochu used for preparation. At this time, in the present invention, koji may or may not be used, but if koji is used, the enzyme amount must be changed appropriately taking into consideration the enzyme in the koji. The brewing process is the same as general mirin brewing, starting from 25:00 at the time of brewing.
The temperature is set as high as 30℃, then gradually lowered to room temperature, and brewing is completed in about 40 to 60 days. The mirin obtained here contains a large amount of non-fermentable oligosaccharides and is rich in flavor. Next, methods for measuring the activity of each enzyme used in the present invention will be explained. 1 α-Amylase: 0.5ml of enzyme solution in 1% soluble starch solution (PH5.0)
Add 0.0317 g of iodine, 0.1 g of potassium iodide, and 10% hydrochloric acid to 1 and perform an enzyme reaction at 40℃.
Add 0.1ml of enzyme reaction solution to 10ml of iodine solution containing 50ml.
was added, this coloration was measured using a 10 mm cell at a wavelength of 670 nm, and the enzyme reaction time (t) at which the transmittance (T) was 66% was determined, and D ⁴⁰ ₃₀ = 10 ml (starch/solution) x 1 The Holgems value was calculated using the formula: /0.5 (enzyme amount) x 30/t, and was taken as an activity unit. That is, 1 unit was defined as the activity of 1 ml of enzyme solution decomposing 1 ml of 1% soluble starch solution in 30 minutes at 40°C. 2 Transglucosidase: 0.5ml of 4% α-methyl-D-glucoside solution
Add 0.5 ml of the enzyme solution to a mixture of 0.02 M acetate buffer (PH 0.5) and 0.5 ml, and perform the enzyme reaction at 40°C for 60 minutes. Add 0.5 ml of 0.1N sodium hydroxide solution to stop the reaction. After 2 hours, add 0.5ml of 0.1N hydrochloric acid solution to neutralize.
Add 2 ml of glucostat reagent to this and heat at 30℃.
Allow the enzyme reaction to occur for 15 minutes, then add one drop of 4N hydrochloric acid to stop the reaction. Separately, as a control, a sample of 0 minutes of enzyme reaction is operated in the same manner and color is developed.
Measure the absorbance with a 10 mm cell at a wavelength of 400 nm, and add 2 ml of glucostat reagent to 2.5 ml of glucose solution.
was added and the same operation was performed to determine the amount of glucose produced in the enzyme reaction solution from the colored calibration curve and use it as an activity unit. In other words, 1 ml of enzyme solution is
One unit was defined as the activity of producing 1 μg of glucose in 60 minutes at 40°C. 3. Acidic protease: Add 0.5 ml of enzyme solution to a mixture of 1.5 ml of 2% milk casein solution (PH3.0) and 1.0 ml of Matsukurubain buffer (PH3.0), and let it act for 60 minutes at 38°C. , 3 ml of 0.4M trichloroacetic acid solution was added to stop the reaction, and the precipitate was separated. Add 5 ml of 0.4M sodium carbonate solution and 1 ml of phenol reagent to 1 ml of the solution, and perform color development at 38°C for 30 minutes. Separately, as a control, a sample with 0 minutes of enzyme reaction was operated in the same manner and color was developed. 660nm
The difference in absorbance between the test solution and the control solution was determined from a calibration curve created by adding 5 ml of 0.4 M sodium carbonate solution and 1 ml of phenol reagent to 1 ml of tyrosine solution and developing the color at 38°C for 30 minutes. Converting to the amount of tyrosine, 1ml of enzyme is 38℃
The activity of producing 1 μg of tyrosine in 60 minutes was defined as 1 unit. Next, the present invention will be specifically explained using test examples and examples. Test example 1 Koji ratio in 4 stages: 0, 5, 10, 15%, transglucosidase addition at 0, 60, 125, 250 units/
A total of 16 groups of rice prepared in 4 stages of g polished rice.
The test was carried out on a scale of 150 g and 135 ml of 30% alcohol. The product temperature is 30℃ for the first 10 days, and then
Aging was performed for 55 days at 18-20°C. The mash was centrifuged at 3000 rpm for 30 minutes, and the supernatant was removed to make mirin. The liquid volume and amount of non-fermentable sugar produced were investigated. The results are shown in Table 1.

[Table] As is clear from Table 1, the production of non-fermentable oligosaccharides does not increase even if transglucosidase is added in normal koji preparation, but in moromi without using koji, a large amount of non-fermentable oligosaccharides is produced. I understand that. It also appears that unless the use of koji is kept to at most 5%, production of large amounts of non-fermentable oligosaccharides cannot be expected. The appropriate amount of transglucosidase to be added was 100 to 300 units/g of polished rice. Test Example 2 100g of glutinous rice was soaked and steamed in a conventional manner, and 15000 units of α-amylase (150 units/g white rice) was added to it.
Add 90ml of 30% alcohol containing 10,000 units of transglucosidase (100 units/g white rice) to make 4 mash, and add transglucosidase to a mixture of 90g of glutinous rice and 10g of rice malt.
90 ml of 30% alcohol containing 10,000 units was added to make four bottles of mash. To these, acidic protease was added in units of 0, 10,000 (100 units/g white rice), 20,000 (200 units/g white rice), and 40,000 (400 units/g white rice), respectively, and incubated at 30°C for 8 days at 20°C. The wine was aged for 16 days. The amounts of α-amylase and transglucosidase in the koji preparation were 120 and 180 units/g polished rice, respectively. In addition, the amount of acidic protease brought in from koji was 180 units/g of polished rice. The mash was centrifuged at 12,000 rpm for 20 minutes, and the supernatant was removed to make mirin. The liquid volume and the amount of non-fermentable oligosaccharide produced were investigated. The results are shown in Table 2.

[Table] As is clear from Table 2, it is clear that the addition of acidic protease promotes the production of non-fermentable oligosaccharides. The amount added is 1g of total rice.
200 to 600 units was appropriate. Note that when koji is used, the amount of non-fermentable oligosaccharides produced is much smaller than when only enzymes are used. Therefore, in order to produce a large amount of non-fermentable oligosaccharides, it is desirable to have a koji ratio of 5% or less and to create a koji with less glucoamylase. From the above results, the enzyme formulation for producing mirin, which is the objective of the present invention, is as follows:
It is appropriate that α-amylase should be 100 to 300 units, transglucosidase should be 100 to 300 units, and acid protease should be 200 to 600 units. Example 1 1,500 g of glutinous rice was soaked and steamed in a conventional manner, and this was mixed with α-amylase, [biodiastase (manufactured by Amano Pharmaceutical Co., Ltd.)] 376,000 units (250 units/g glutinous rice), acidic protease [Morsin (Mori Shin)] Manufactured by Pharmaceutical Company)]
750,000 units (500 units/g glutinous rice), transglucosidase (manufactured by the purification method described above)
30% containing 180000 units (120 units/g sticky rice)
1350ml of alcohol was added to prepare the mash. The moromi was aged at 30°C for 7 days and at 18-20°C for 30 days. As a control, 1280 g of glutinous rice and 220 g of rice malt
A moromi was prepared using the same method, and the fermentation was carried out in the same manner. The aged moromi was pressed using a small squeezer to obtain mirin. Table 3 shows the ingredients of the produced mirin.

【table】

[Table] Next, the two mirin produced by the method of the present invention and the conventional method were subjected to a three-point discrimination test. Judgments were made on 6 sets of samples by a panel of 4 people. In other words, the number of correct answers out of 24 judgments was 20, so it is 0.1%.
It was determined that the mirin manufacturing method of the present invention had a significant difference with a risk rate of . In terms of taste characteristics, it was determined that the mirin of the present invention had a smoother and sweeter taste.

Claims (1)

[Claims] 1. A novel method for producing mirin, which comprises adding an enzyme agent containing transglucosidase, α-amylase, and acidic protease as is or substituting part or all of rice malt for mirin production.