JP2023039161A - Method for producing high-protein soybean meal - Google Patents

Abstract

To provide a method for producing high-protein soybean meal with excellent nutritional properties, and to provide a method for producing high-protein soybean meal that can also produce industrially valuable ethanol when producing the high-protein soybean meal.SOLUTION: A method for producing high-protein soybean meal is characterized by mixing soybean meal and water and subjecting the resulting mixture to saccharification treatment and alcoholic fermentation treatment, where cellulase is added for the saccharification treatment and microorganisms are added for the alcoholic fermentation treatment. The method for producing high-protein soybean meal is characterized in that the saccharification treatment and alcoholic fermentation treatment are performed in the same step.SELECTED DRAWING: None

Description

The present invention relates to a method for producing high protein soybean meal.

Soybean meal has been widely used as a raw material for feed.
For example, 1 to 40 parts by mass of starch, 10 to 80 parts by mass of soybean meal, and 1 to 30 parts by mass of oil are molded in order to provide poultry with the necessary nutrients and achieve a sufficient water absorption effect. Therefore, a poultry feed has been developed that is fed while maintaining its particle shape by absorbing water before use (Patent Document 1).
In particular, since soybean meal has a high crude protein content, it has often been added to feeds to make feeds rich in nutritional value.
For example, by using soybean meal instead of a binder in solid feed for fish farming, not only does the solid feed have a moderate hardness that is easy for fish to ingest, but the solid feed has a uniform particle size and a crude protein content. It was found to be relatively high and rich in nutritional value (Patent Document 2).
In addition, by using dehulled soybean meal, which has about 10% higher crude protein content, as the soybean meal component in livestock feed, the amount of soybean meal required to secure the same crude protein content can be reduced. It is reported that it can be reduced by about 10% (Patent Document 3).

JP 2005-295814 Japanese Patent Laid-Open No. 05-076291 JP 2004-113106

It is an object of the present invention to provide a method for producing high protein soybean meal with good nutritional properties.
Another object of the present invention is to provide a method for producing high-protein soybean meal that can also produce industrially valuable ethanol when producing high-protein soybean meal.

In order to solve the above problems, the present inventors conducted extensive studies and found that the ratio of crude protein in soybean meal can be increased by conducting saccharification treatment and alcoholic fermentation treatment in the same process. rice field. In addition, the inventors have found that ethanol productivity can be enhanced by conducting saccharification treatment and alcoholic fermentation treatment in the same step, and have completed the present invention.

Specifically, the present invention includes the following aspects.
[1] Characterized by mixing soybean meal and water, subjecting the resulting mixture to saccharification treatment and alcoholic fermentation treatment, adding cellulase for the saccharification treatment, and adding microorganisms for the alcoholic fermentation treatment. A method for producing a high protein soybean meal.
[2] The production method according to [1], wherein the saccharification treatment and the alcoholic fermentation treatment are performed in the same step.
[3] When the ratio of the crude protein content to the carbohydrate content of the raw material soybean meal (crude protein content / carbohydrate content) is 100, the ratio of the crude protein content to the carbohydrate content of the high-protein soybean meal obtained (crude protein content /carbohydrate content) is 130-160.
[4] The high protein according to any one of [1] to [3], wherein the mixture of soybean meal and water is not sterilized before the saccharification treatment and/or alcoholic fermentation treatment. A method for producing soybean meal.
[5] The alcohol fermentation treatment according to any one of [1] to [4], wherein the amount of ethanol produced per 100 g of dry weight of soybean meal is 3.5 to 10 g. A method for producing a high protein soybean meal.
[6] The method for producing high-protein soybean meal according to any one of [1] to [5], wherein the saccharification treatment and the alcoholic fermentation treatment are performed by a solid-state fermentation method.
[7] The production according to any one of [1] to [6], characterized in that the amount of water in the reaction system in which the saccharification treatment and the alcohol fermentation treatment are performed is 40 to 60% by mass. Method.
[8] The method for producing high-protein soybean meal according to any one of [1] to [7], wherein the same step is performed at 25 to 45°C.
[9] A feed containing high-protein soybean meal produced by the method for producing high-protein soybean meal according to any one of [1] to [8].

Ethanol can also be obtained in the production of high protein soybean meal by practicing the present invention. By using the obtained high-protein soybean meal as a feed material, the crude protein content in the feed can be increased. When the high-protein soybean meal of the present invention is used as a raw material for feed, feed with increased energy value can be fed to aquatic products such as livestock, fish, and crustaceans.
In addition, the obtained ethanol can be used for various purposes as bioethanol.

(soybean meal)
Soybean meal is soybean meal that is made by evaporating the organic solvent after an oil extraction process that extracts the oil content of soybeans using an organic solvent such as n-hexane, or extracts the oil content from soybeans with a press. It is the soybean meal obtained after grinding and contains 8 to 15% by mass of water.
The amount of soybean meal is preferably 40 to 60 mass%, more preferably 55 to 45 mass%, and still more preferably 52 to 48 mass% as a dry weight in the total reaction system in which saccharification treatment and alcoholic fermentation treatment are performed. preferably used.

(water, other additives)
Water in the present invention is not particularly limited, and may be, for example, distilled water, pure water, tap water, sterilized water, or physiological saline.
Water is preferably used in an amount of 40 to 60% by mass, more preferably 45 to 55% by mass, and even more preferably 48 to 52% by mass in the total reaction system in which saccharification treatment and alcoholic fermentation treatment are performed. .
Such a water content is preferable because it becomes a solid fermentation method.
Here, the term "solid fermentation method" refers to a fermentation method in which the water content is set lower than that of the liquid fermentation method, and little waste liquid is discharged not only during the fermentation period but also after the fermentation is completed.

0 to 10 parts by mass of additives such as minerals, sugars, amino acids, medium components, enzymes and microorganisms may be added to 100 parts by mass of the mixture of soybean meal and water before saccharification and alcoholic fermentation. Specific examples of minerals include phosphorus, magnesium, sodium, potassium, iron and the like. Specific examples of sugars include sucrose, glucose, maltose, oligosaccharides and the like. Specific examples of amino acids include glutamic acid, lysine, and the like, and may be crude protein constituent amino acids, non-constitutive amino acids such as GABA, or crude protein hydrolysates. Specific examples of medium components include yeast extract, malt extract, peptone, and the like.
Specific examples of enzymes for saccharification include cellulase, hemicellulase, glucosidase, or mixtures thereof. In the saccharification treatment of the present invention, the cellulase (fiber-degrading enzyme) is cellulase produced by Acremonium genus microorganisms, particularly cellulase produced by Acremonium cellulotycus (product name: Acremonium cellulotycus). Remocellulase KM, manufactured by Kyowa Kasei Co., Ltd.), or cellulase produced by Aspergillus genus microorganisms, especially cellulase produced by Aspergillus niger (product name: Cellulase, from Aspergillus niger, MP Biomedicals, Inc.) .), and cellulase produced by Aspergillus niger (product name: Cellulase from Aspergillus niger, manufactured by Tokyo Chemical Industry Co., Ltd.) are used. In the present invention, it is preferable to use cellulase produced by Acremonium celluloticus.
Specific examples of microorganisms for alcoholic fermentation include filamentous fungi, yeasts such as Saccharomyces cerevisiae, Shizosaccharomyces pombe, Pichia stipitis, and Zymomonas mobilis. Yeast such as Saccharomyces cerevisiae is preferably used for the alcoholic fermentation treatment of the present invention, and yeast that can be used for solid-state fermentation is particularly preferred.
Hereinafter, water or water and these additives are collectively referred to as "water or the like". Among these additives, water-soluble ones may be added after being solubilized in the water.

(mixture)
The mixing of soybean meal and water or the like in the present invention means mixing soybean meal and water or the like manually or with a device. The more uniform the distribution of water in the solid content after mixing, the better the efficiency of the saccharification treatment or the alcoholic fermentation treatment. be. In general, it has been considered desirable to sterilize a mixture of soybean meal and water, etc., in an autoclave to prevent the growth of microorganisms other than the desired microorganisms during saccharification or alcoholic fermentation. It was found that the present invention can omit the sterilization treatment before the saccharification treatment and the alcoholic fermentation treatment.

(saccharification treatment)
The saccharification treatment in the present invention is performed by adding 0.01 to 1.0% by weight of a fiber-degrading enzyme to the fiber (cellulose etc.) contained in soybean meal to a mixture of soybean meal and water etc. It refers to breaking down the fiber contained in soybean meal into glucose. As the fiber-degrading enzyme used for saccharification, cellulase can be used alone, or cellulase can be used in combination with an enzyme such as hemicellulase or glucosidase.
The amount of cellulase used is preferably 0.01 to 6% by weight based on the mixture of soybean meal and water. Further, it is more preferable to add 0.05 to 2% by weight, more preferably 0.05 to 1% by weight.
Other than that, the enzyme used for the saccharification treatment may be any enzyme that can decompose the fiber, and even if it is a commercial product, a culture solution in which filamentous fungi are cultured, or a further refined product, the filamentous fungus itself can be used. It can be.

(alcohol fermentation treatment)
The alcoholic fermentation treatment in the present invention refers to a process in which a mixture of soybean meal and water or the like or a saccharified mixture of soybean meal and water or the like is inoculated with microorganisms to produce ethanol by the microorganisms. . Regarding the microorganisms to be used, yeasts such as Saccharomyces cerevisiae, Shizosaccharomyces pombe, and Pichia stipitis can be used. Anything can be used, including genetically modified ones. Microorganisms that have been preserved by slant or freezing may be used, but when using Saccharomyces cerevisiae, commercially available baker's yeast may be used. When using slant- or frozen-preserved strains, it is desirable to pre-culture them in a liquid medium before use and use the pre-culture solution or the like. The liquid medium used for the pre-culture may be one suitable for culturing yeast or bacteria, such as 1% by mass yeast extract, 2% by mass peptone, and 3% by mass glucose.
In the present invention, yeast such as Saccharomyces cerevisiae is preferably used as the microorganism used for alcoholic fermentation, and yeast that can be used for solid fermentation is particularly preferable. In addition, the amount of the yeast used is a yeast suspension prepared so that the absorbance (hereinafter referred to as OD) when measured at 660 nm is 0.2, and the total reaction system (yeast (suspension) + It is preferable to add so that the amount of yeast present in the meal + added water is OD = 0.05 to 0.15. Further, it is more preferable to add so that OD=0.05 to 0.1.

(When performing saccharification and alcoholic fermentation in separate processes or in the same process)
In the present invention, a mixture containing enzymes, microorganisms, soybean meal, water, etc. is subjected to saccharification treatment and alcoholic fermentation treatment, but saccharification treatment and alcoholic fermentation treatment may be performed in separate steps (" It is preferable to use the same process (referred to as "multiple parallel fermentations") because the amount of alcohol produced increases when the same process is used.
In addition, performing in a separate process is performing an alcoholic fermentation process, after performing a saccharification process, for example.
When the saccharification treatment and the alcohol fermentation treatment are performed in separate steps, the saccharification temperature is preferably 40 to 60°C, more preferably 42 to 58°C, and even more preferably 45 to 55°C. On the other hand, the fermentation temperature is preferably 20 to 40°C, more preferably 22 to 38°C, even more preferably 25 to 35°C.
Further, when the saccharification treatment and the alcohol fermentation treatment are performed in the same step, the treatment temperature is preferably 25 to 45°C, more preferably 27 to 43°C, and further preferably 30 to 40°C. preferable.
The saccharification treatment time and the alcohol fermentation treatment time (the total treatment time when the saccharification treatment and the alcohol fermentation treatment are performed in separate steps) are preferably 4 to 180 hours, more preferably 72 to 180 hours. , more preferably 24 to 180 hours.

(After solid-liquid separation, drying and distillation)
A mixture of soybean meal and water, subjected to saccharification treatment and alcoholic fermentation treatment, is subjected to solid-liquid separation and dried as necessary. The result is called high protein soybean meal. Drying may be dry heat drying, vacuum drying, freeze drying, spray drying, or the like, as long as the moisture and ethanol of the mixture obtained by saccharification treatment and/or alcohol fermentation treatment can be evaporated. If ethanol is recovered by steam distillation or the like before drying, the recovered ethanol can be used for industrial purposes or fuel.
The ethanol production amount in the present invention is preferably 3.5 to 10 g, more preferably 3.8 to 9 g, and 4 to 8 g per 100 g of dry weight of soybean meal. is more preferred.
It is preferable that the saccharification treatment and the alcoholic fermentation treatment are performed by a solid-state fermentation method. The use of the solid-state fermentation method eliminates the need for the above-described solid-liquid separation and is preferable because no waste liquid is generated.

(high protein soybean meal)
The high-protein soybean meal in the present invention is a high-protein soybean meal processed product in which the crude protein content is increased compared to the raw material soybean meal by subjecting soybean meal to saccharification treatment and alcoholic fermentation treatment, and the crude protein content is high. It is 55 to 65% by mass, preferably 55 to 62% by mass in the dry matter of protein soybean meal.
For example, if soybean meal containing about 50% by mass of crude protein in dry matter is used as a raw material to produce the present invention, high-protein soybean meal with a crude protein content of about 58% by mass in dry matter can be obtained. can be done.
Further, for example, if the high-pro soybean meal of the present invention having a high crude protein content of about 54% by mass in the dry matter is used as a raw material to produce the present invention, high-protein soybeans with a crude protein content of about 62% by mass in the dry matter You can get meal.
Also, when the ratio of the crude protein content to the carbohydrate content of the raw material soybean meal (crude protein content / carbohydrate content) is 100, the ratio of the crude protein content to the carbohydrate content of the high protein soybean meal obtained (crude protein content / carbohydrate content) is preferably 130-160, more preferably 135-155, even more preferably 140-150.
In the production method of the present invention, the more carbohydrates (crude fiber, soluble nitrogen-free substances (starch, sugar, etc.)) in the raw material soybean meal are consumed by saccharification treatment and alcoholic fermentation treatment, the more the amount of carbohydrates is reduced, Higher values of crude protein/carbohydrate ratio, ie higher percentage of crude protein in high protein soybean meal.
The crude protein content referred to herein refers to a value obtained by multiplying the total nitrogen obtained by the Kjeldahl method by 6.25.
A mixed feed using the high-protein soybean meal produced by the production method of the present invention can be used as a feed without any problem.

(feed)
The high-protein soybean meal obtained by the production of the present invention can be mixed with feed ingredients and feed additives containing sugar, protein, amino acids, fiber, minerals, oil, antibacterial components, etc., and used as feed. can.
Specific examples of feed ingredients and feed additives include corn, sorghum, corn gluten field, corn starch, rice bran, ordinary soybean meal, high-pro soybean meal, rapeseed meal, rapeseed meal saccharified and alcohol-fermented, wheat bran, and oats. , milk casein, whey, fish meal, various vitamins, vitamin mixes, mineral mixes, amino acid preparations, calcium carbonate, monocalcium phosphate, vegetable oils and fats, animal oils and fats, salt and the like.
When the total amount of feed is 100% by mass, the content of high-protein soybean meal in the feed is preferably 0.1 to 30% by mass, more preferably 0.5 to 20% by mass, and 1 to It is more preferably 10% by mass.
Further, for example, when using plant meal as a protein source as a feed ingredient, the high protein soybean meal obtained by the production of the present invention can be used as it is. , high-pro soybean meal, rapeseed meal, rapeseed meal subjected to saccharification treatment and alcoholic fermentation treatment, etc. can also be used.
A feed containing high-protein soybean meal can be produced by mixing the high-protein soybean meal produced by the production method of the present invention with the various feed ingredients and feed additives described above. Mixing can be performed using a mixer such as a ribbon mixer, a V-type mixer, or a W-type mixer.

EXAMPLES In order to describe the present invention in more detail, examples are shown below, but the present invention is not limited to these.

<Soybean meal used in the experiment>
Two types of soybean meal were used in this experiment: regular soybean meal and high-pro soybean meal, which has a higher crude protein content than regular soybean meal. Table 1 shows the component analysis values of the soybean meal used.

<Example 1: Solid-state fermentation method for soybean meal (small-scale test)>
100 g of soybean meal or high-pro soybean meal was placed in a reaction vessel as a dry weight, and after adding 0.1% by mass of various fiber-degrading enzymes (cellulase) shown in Table 2, a suspension of yeast (Saccharomyces cerevisiae NBRC-0203) was added. was added such that the total reaction system had an OD=0.1. After that, water was added so that the total water content of the reaction system was 50% by mass, and about 200 g of a mixture of soybean meal or high-pro soybean meal and water was obtained.
These were subjected to solid state fermentation at 37° C. for 7 days without autoclave sterilization.
The yeast (Saccharomyces cerevisiae) used was Saccharomyces cerevisiae NBRC-0203. NBRC-0203 can be obtained by distribution from the Depository of National Institute of Technology and Evaluation (NBRC).
After 3 days and 7 days, the solid portion was collected, weighed, and then the concentration of ethanol contained in the solid portion was measured. The ethanol concentration was measured by adding the same amount of water to the solid portion, centrifuging at 1500 rpm for 10 minutes, and collecting the supernatant. Furthermore, it is centrifuged at 15000 rpm for 10 minutes, the supernatant is collected, diluted 1000 times with water, and ethanol analysis kit (product name: The F-kit, sold by JK International Co., Ltd.) Concentration was measured. Table 3 shows the results.

In Table 3, three types of enzymes were used as fiber-degrading enzymes, and all enzymes detected a sufficient concentration of ethanol. In particular, enzyme KM detected high values in both soybean meal and high-pro soybean meal. Enzyme KM is an endoglucanase produced by Acremonium genus microorganisms, and it is considered that the high β-glucosidase activity influences such good results. Therefore, the following experiment was conducted by focusing on the enzyme KM, which is produced by Acremonium genus microorganisms, as the fiber-degrading enzyme.

<Example 2: Solid-state fermentation method for soybean meal and evaluation of high-protein soybean meal (medium-scale test)>
1000 g of soybean meal or high-pro soybean meal was placed in a reaction vessel as a dry weight, and after adding 0.1% by mass of the enzyme KM shown in Table 2, a suspension of yeast (Saccharomyces cerevisiae NBRC-0203) was added to the total reaction system. It was added so that OD=0.1. After that, water was added so that the total water content of the reaction system was 50% by mass, and about 2000 g of a mixture of soybean meal or high-pro soybean meal and water was obtained.
These were subjected to solid state fermentation at 37° C. for 7 days without autoclave sterilization.
In order to analyze various components of the obtained high-protein soybean meal, it was subjected to heat treatment under reduced pressure (72° C., −0.1 MPa) for 8 hours. After that, the dried high-protein soybean meal was pulverized with a pulverizer (manufactured by Osaka Chemical Co., Ltd., apparatus "Absolute mill ABS-W") to obtain an analysis sample.
The amount of ethanol produced per 100 g dry weight of soybean meal or high-pro soybean meal in the medium-scale test, and each component of soybean meal or high-pro soybean meal in the medium-scale test (moisture, crude protein, crude fat, crude ash, carbohydrates (crude Fiber, soluble nitrogen-free substances)) and the ratio of the crude protein content to the carbohydrate content (crude protein content / carbohydrate content) of the raw soybean meal or high-pro soybean meal is 100, the resulting high protein soybean meal The ratio of crude protein content to carbohydrate content (crude protein content/carbohydrate content) is shown in Tables 4 and 5. Analytical values for crude protein, crude fat, crude ash, and carbohydrate (crude fiber and soluble nitrogen-free) contents are in dry matter.
The amount of ethanol produced per 100 g of dry weight of soybean meal was measured by the same analytical method as described in Example 1.
The water content of the obtained high-protein soybean meal was determined by normal pressure heating and drying, the crude protein was determined by the Kjeldahl method, the crude fat was determined by the Soxhlet method, the crude ash was determined by direct incineration, the crude fiber was determined by filtration, and the soluble nitrogen-free substances were determined by 100%. It was calculated as a value obtained by subtracting moisture, crude protein, crude fat, crude ash, and crude fiber from the Carbohydrates were calculated as the sum of crude fiber and soluble nitrogen-free substances.

<Comparative Example 1: Solid Fermentation Method for Rapeseed Meal (Medium Scale Test)>
Instead of soybean meal, 3000 g of rapeseed meal as a dry weight was placed in a reaction vessel, and 0.1% by mass of enzyme KM shown in Table 2 was added. was added so that OD=0.1. After that, water was added so that the total water content of the reaction system was 50% by mass, and about 6000 g of a mixture of rapeseed meal and water was obtained.
These were subjected to solid state fermentation at 37° C. for 7 days without autoclave sterilization.
The amount of ethanol produced per 100 g dry weight of rapeseed meal was measured by the same analytical method as described in Example 1. Further, the analytical values of each component of the rapeseed meal after fermentation and the ratio of crude protein content/carbohydrate content were measured by the same analytical method as described in Example 2. These analysis results are shown in Table 6.

As can be seen from Tables 4 and 5, ethanol was produced so that the amount of ethanol produced per 100 g dry weight of soybean meal or high-pro soybean meal was 5.2 to 6.7 g. From this result, it was found that even the medium-scale test can produce ethanol equal to or greater than that of the small-scale test.
Further, as can be seen from Tables 4 and 5, the crude protein content of soybean meal before and after fermentation decreased from 50.03% by mass to 58.16% by mass, and the crude protein content of Hypro soybean meal decreased from 54% by mass to 54% by mass. It improved from 31 mass % to 61.50 mass %. This is due to the conversion of carbohydrates (crude fiber, soluble nitrogen-free substances (starch, sugars, etc.)) in soybean meal or high-pro soybean meal to ethanol by fermentation, which is relatively contained in soybean meal or high-pro soybean meal. It shows that the crude protein content was improved.
On the other hand, the carbohydrate content of soybean meal before and after fermentation decreased from 40.61% to 31.33% by mass, and the carbohydrate content of HyPro soybean meal decreased from 36.56% to 28.67% by mass. . This indicates that fermentation has converted the carbohydrates (crude fiber, soluble nitrogen-free substances (starch, sugars, etc.)) of soybean meal or high-pro soybean meal to ethanol.
Also, when the ratio of crude protein content to carbohydrate content on the 7th day of fermentation is taken as 100, the ratio of crude protein content to carbohydrate content of high protein soybean meal was 150.7 for soybean meal and 144.0 for high-pro soybean meal. became 4. This is because carbohydrates (crude fiber, soluble nitrogen-free substances (starch, sugar, etc.)) in the raw material soybean meal are consumed by saccharification and alcoholic fermentation, resulting in a decrease in the amount of carbohydrates and a decrease in crude protein/carbohydrate content. It is thought that the ratio has increased.
In addition, the ratio of crude protein content to carbohydrate content of rapeseed meal subjected to saccharification treatment and alcoholic fermentation treatment was 126.8 when the ratio of crude protein content to carbohydrate content on the 7th day of fermentation was 100 (Table 6).
Looking at the analysis values of carbohydrates (crude fiber, soluble nitrogen-free substances (starch, sugar, etc.)) this time, the raw material soybean meal was 40.61% by mass, and the raw material high-pro soybean meal was 36.56% by mass. Rapeseed meal was 46.00% by mass, which was higher than each raw material soybean meal. Therefore, raw rapeseed meal contains more carbohydrates (crude fiber, soluble nitrogen-free substances (starch, sugar, etc.)), which are fermentation substrates.
Expected to consume more carbohydrates (crude fiber, soluble nitrogen-free (starches, sugars, etc.)) than raw soybean meal, resulting in higher protein content and higher ratio of crude protein content to carbohydrate content. Was.
However, contrary to expectations, the ratio of crude protein content to carbohydrate content was higher when soybean meal containing less carbohydrates (crude fiber, soluble nitrogen-free substances (starch, sugar, etc.)) was used as the raw material. In other words, it was an unexpected result considering that the carbohydrate content of rapeseed meal was higher than that of soybean meal, which is the fermentation substrate.
From this experiment, by using soybean meal or high-pro soybean meal as a raw material and performing saccharification treatment and alcoholic fermentation treatment, it was possible to increase the ratio of crude protein content more than when rapeseed meal was used as a raw material. have understood.

<Example 3: Solid-state fermentation method for soybean meal with varying amounts of fiber-degrading enzyme added (small-scale test)>
100 g of soybean meal or high-pro soybean meal was placed in a reaction vessel as a dry weight, and after adding the enzyme KM shown in Table 2 in the range of 0.01 to 5.0% by mass, yeast (Saccharomyces cerevisiae NBRC-0203) was suspended. Liquids were added such that the total reaction system had an OD=0.1. After that, water was added so that the total water content of the reaction system was 50% by mass, and about 200 g of a mixture of soybean meal or high-pro soybean meal and water was obtained.
These were subjected to solid state fermentation at 37° C. for 7 days without autoclave sterilization.
The amount of ethanol produced per 100 g of dry weight of soybean meal was measured by the same analytical method as described in Example 1. Table 7 shows the results.

From the results in Table 7, there was a tendency that the greater the amount of enzyme added, the greater the amount of ethanol produced per 100 g of dry weight of soybean meal.

<Comparative Example 2: Test in which saccharification treatment and alcoholic fermentation treatment were performed separately (small-scale test)>
Put 100 g of soybean meal or high-pro soybean meal as a dry weight in a reaction vessel, add 0.1% by mass of the enzyme KM shown in Table 2, and then add water so that the total water content of the reaction system is 50% by mass. About 200 g of a mixture of soybean meal or hypro soybean meal and water was obtained.
These were subjected to saccharification treatment at 50° C. for 3 days without autoclave sterilization.
Thereafter, a suspension of yeast (Saccharomyces cerevisiae NBRC-0203) was added so that the total reaction system had an OD of 0.1, and alcoholic fermentation was performed at 37° C. for 3 days.
The amount of ethanol produced per 100 g of dry weight of soybean meal was measured by the same analytical method as described in Example 1. Table 8 shows the results.

According to Table 4, the saccharification treatment was performed at 50 ° C., which is the optimum temperature for the enzyme KM, and then the fermentation treatment was performed. Hypro soybean meal: 4.05 g) yielded less ethanol. From this, it was found that the parallel multiple fermentation in which the saccharification treatment and the fermentation treatment are performed simultaneously has higher ethanol production efficiency than the method in which the saccharification treatment and the fermentation treatment are performed separately.

<Example 4: Ethanol solid-state fermentation method of soybean meal with various yeasts (small-scale test)>
100 g of soybean meal or high-pro soybean meal in terms of dry weight was placed in a reaction vessel, and after adding 0.1% by mass of the enzyme KM shown in Table 2, suspensions of various yeasts shown in Table 9 were added to the reaction vessel so that the total reaction system was OD. = 0.1. After that, water was added so that the total water content of the reaction system was 50% by mass, and about 200 g of a mixture of soybean meal or high-pro soybean meal and water was obtained.
These were subjected to solid state fermentation at 37° C. for 7 days without autoclave sterilization.
The amount of ethanol produced per 100 g of dry weight of soybean meal was measured by the same analytical method as described in Example 1. Table 9 shows the results.

From the results in Table 10, it was found that the same amount of ethanol was produced even when the yeast strain was changed.

Claims (9)

Soybean meal and water are mixed, the resulting mixture is subjected to saccharification treatment and alcoholic fermentation treatment, cellulase is added for the saccharification treatment, and microorganisms are added for the alcoholic fermentation treatment. A method for producing a high protein soybean meal. 2. The production method according to claim 1, wherein the saccharification treatment and the alcoholic fermentation treatment are performed in the same step. When the ratio of the crude protein content to the carbohydrate content of the raw material soybean meal (crude protein content / carbohydrate content) is 100, the ratio of the crude protein content to the carbohydrate content of the high protein soybean meal obtained (crude protein content / carbohydrate content ) is 130-160. The method for producing high-protein soybean meal according to any one of claims 1 to 3, wherein the mixture of soybean meal and water is not sterilized before the saccharification treatment and/or alcoholic fermentation treatment. . 5. The high-protein soybean meal according to any one of claims 1 to 4, characterized in that the amount of ethanol produced per 100 g of dry weight of soybean meal is 3.5 to 10 g in the alcoholic fermentation treatment. Production method. The method for producing high-protein soybean meal according to any one of claims 1 to 5, wherein said saccharification treatment and said alcoholic fermentation treatment are performed by a solid-state fermentation method. Production of the high protein soybean meal according to any one of claims 1 to 6, characterized in that the amount of water is 40 to 60% by mass in the reaction system in which the saccharification treatment and the alcoholic fermentation treatment are performed. Method. A process for producing high protein soybean meal according to any one of claims 1 to 7, characterized in that said same step is carried out at 25-45°C. A feed containing high-protein soybean meal produced by the method for producing high-protein soybean meal according to any one of claims 1 to 8.