JP7669096B2 - Method for producing high protein rapeseed meal - Google Patents

Description

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

The residue left over after the oil has been removed from oilseeds such as rapeseed and soybeans is primarily used as animal feed as a source of protein. However, rapeseed meal, which is the residue left over from the extraction of rapeseed, has the disadvantage that it has a lower protein content than soybean meal and is high in fiber, resulting in a low energy value for livestock.

To solve these problems, several methods have been proposed to increase the protein content in rapeseed meal. For example, a method has been disclosed in which rapeseed meal is separated using a sieve to separate it into a high-protein fraction and a low-protein fraction, but the low-protein fraction has low feed value and is therefore of limited use (Patent Document 1). A method has also been disclosed in which rapeseed is peeled, then oiled and the rapeseed meal is recovered, but this method, which peels rapeseed with small grain sizes, is expensive and unrealistic (Non-Patent Document 1). A method has also been disclosed for producing high-protein, low-glucosinolate rapeseed meal by mixing rapeseed meal with water, subjecting the mixture to saccharification and/or alcohol fermentation, separating the mixture into solid and liquid, and drying (Patent Document 2), but further improvements in ethanol production capacity, etc., have been desired.

Patent No. 3970917 Patent No. 5601763 W. Grala, et al., Real apparent protein and amino acid digestibilities and endogenous nitrogen losses in pigs fed soybean and rapeseed products. , Journal of Animal science, 76, p. 769-p. 577 (1998)

The objective of the present invention is to improve the ethanol production capacity in a method for producing high-protein rapeseed meal that can produce industrially valuable ethanol and simultaneously produce high-protein rapeseed meal with excellent nutritional properties.

The inventors conducted extensive research to solve the above problems and discovered that ethanol production can be improved by using cellulase produced by Acremonium microorganisms as a fibrous decomposition enzyme for saccharification, thus completing the present invention.

Specifically, the present invention may include the following aspects.
[1] A method for producing a high-protein rapeseed meal, which comprises mixing rapeseed meal with water, subjecting the resulting mixture to saccharification and alcoholic fermentation, adding a fibrous enzyme for the saccharification, and adding a microorganism for the alcoholic fermentation, wherein cellulase produced by Acremonium cellulolyticus is used as the fibrous enzyme , and the mixture of rapeseed meal and water is not sterilized before the saccharification and/or alcoholic fermentation .
[2] The method according to [1], characterized in that in the alcohol fermentation treatment, the amount of ethanol produced per 100 g of dry weight of rapeseed meal is 3.5 g or more.
[3] The method for producing the rapeseed meal according to [1] or [2], characterized in that 40 to 60 parts by weight of the rapeseed meal is mixed with 60 to 40 parts by weight of water.
[4] The method for producing a product according to any one of [1] to [3], characterized in that the saccharification treatment and the alcoholic fermentation treatment are carried out in the same process.
[5] The method according to [4], wherein the same step is carried out at 25 to 45° C.
[6] The method for producing a product according to any one of [1] to [3], characterized in that the saccharification treatment and the alcoholic fermentation treatment are carried out in separate steps.
[7] The method according to [6], wherein the saccharification treatment is carried out at 40 to 60°C, and the alcoholic fermentation treatment is carried out at 20 to 40°C.
[ 8 ] A feed containing high-protein rapeseed meal produced by the production method described in any one of [1] to [7] above.

By carrying out the present invention, it is possible to produce high-protein rapeseed meal at the same time as ethanol. This makes it possible to increase the energy value for livestock. Furthermore, ethanol can also be recovered at a high yield. The ethanol obtained in this way can be used for various purposes as bioethanol. Furthermore, whether the saccharification process and the alcohol fermentation process are carried out in the same process or separately, the bacteriostatic effect of high temperature or alcohol works, making it possible to omit the sterilization process (autoclave process, etc.) that was previously required, and therefore high-protein rapeseed meal can be produced in a simpler manner than before. Furthermore, the use of solid fermentation in the present invention makes it possible to produce an environmentally friendly method of producing high-protein rapeseed meal, since no waste liquid is produced.

FIG. 1 shows ethanol production per 100 g dry weight of rapeseed meal (medium-scale study) (days 7 to 13). FIG. 1 shows the change in rapeseed meal components (protein, total carbohydrate, cellulose) and protein/carbohydrate ratio (medium-scale study) (before fermentation to 10th to 13th days). FIG. 1 shows ethanol production per 100 g dry weight rapeseed meal using various fibrous enzymes at high concentrations (small scale study). FIG. 1 shows the optimum temperature for saccharification (small-scale test) when saccharification and fermentation are separated. FIG. 1 shows the optimum temperature for alcohol fermentation treatment (small-scale test) when saccharification treatment and fermentation treatment are separated. FIG. 1 shows ethanol production per 100 g dry weight of rapeseed meal with different yeasts (small scale studies).

(rapeseed meal)
Rapeseed meal is rapeseed meal produced by extracting rapeseed using a press, then going through an oil extraction process in which the oil remaining in the pressed cake is extracted using an organic solvent such as n-hexane, and then evaporating the organic solvent. Rapeseed meal contains 8 to 15% by mass of moisture.

(Water and other additives)
The water in the present invention is not particularly limited, and may be, for example, distilled water, pure water, or tap water. When rapeseed meal and water are mixed before saccharification or alcohol fermentation, the ratio of rapeseed meal to water is 40 to 60 parts by weight, preferably 45 to 55 parts by weight, rapeseed meal to water 60 to 40 parts by weight, more preferably 48 to 52 parts by weight, rapeseed meal to water 52 to 48 parts by weight. Such a water content is preferable because it is a solid fermentation method. Here, the "solid fermentation method" refers to a fermentation method in which the water content is set lower than that of the liquid fermentation method, and almost no waste liquid is discharged not only during the fermentation period but also after the fermentation is completed.

Additives such as minerals, sugars, amino acids, medium components, enzymes, microorganisms, etc. may be added in an amount of 0 to 10 parts by weight per 100 parts by weight of a mixture of rapeseed meal and water before saccharification and alcohol fermentation. Specific examples of minerals include phosphorus, magnesium, sodium, potassium, iron, etc. Specific examples of sugars include sucrose, glucose, maltose, oligosaccharides, etc. Specific examples of amino acids include glutamic acid and lysine, and may also be protein-constituting amino acids, non-constituting amino acids such as GABA, and protein hydrolysates. Specific examples of medium components include yeast extract, malt extract, peptone, etc.
Specific examples of enzymes for saccharification include cellulase, hemicellulase, glucosidase, or a mixture thereof. In the saccharification treatment of the present invention, as one type of cellulase (fibrous decomposition enzyme), cellulase produced by microorganisms of the genus Acremonium is used, particularly cellulase F produced by Acremonium cellulolyticus (product name: Meiji Acremonium Cellulase F, manufactured by Meiji Seika Pharma Co., Ltd.) and cellulase KM produced by Acremonium cellulolyticus (product name: Acremonium Cellulase KM, manufactured by Kyowa Kasei Co., Ltd.). In the present invention, it is preferable to use cellulase produced by Acremonium cellulolyticus . As described below, it was first discovered in the present invention that ethanol production ability is improved by using cellulases produced by these microorganisms of the genus Acremonium.
Specific examples of microorganisms for alcohol fermentation treatment include filamentous fungi, yeasts such as Saccharomyces cerevisiae, Shizosaccharomyces pombe, and Pichia stipitis, Zymomonas mobilis, etc. For the alcohol fermentation treatment of the present invention, it is preferable to use yeasts such as Saccharomyces cerevisiae, and particularly yeasts that can be used in solid fermentation methods.
In the following description, water or water and these additives will be collectively referred to as "water, etc." Among these additives, water-soluble ones may be solubilized in the water and then added.

(mixture)
In the present invention, the mixing of rapeseed meal with water, etc. refers to mixing the rapeseed meal with water, etc. manually or by device. The more uniform the distribution of moisture in the solids after mixing, the more efficient the saccharification or alcoholic fermentation treatment, and therefore it is desirable, but even if the moisture distribution in the solids is uneven, the purpose of saccharification or alcoholic fermentation can be achieved. In general, it has been considered desirable to sterilize the mixture of rapeseed meal and water, etc., in an autoclave during saccharification or alcoholic fermentation to prevent the proliferation of microorganisms other than the desired microorganisms, but it has been found that the present invention makes it possible to omit the sterilization treatment prior to saccharification or alcoholic fermentation.

(Saccharification process)
The saccharification treatment in the present invention refers to adding a fibrous decomposition enzyme to a mixture of rapeseed meal and water, etc., at 0.01 to 1.0% by weight, to decompose the fibrous material (such as cellulose) contained in the rapeseed meal into glucose. The fibrous decomposition enzyme used in the saccharification treatment can be cellulase alone or cellulase in combination with an enzyme such as hemicellulase or glucosidase, but in the present invention, it is essential to use cellulase produced by a microorganism belonging to the genus Acremonium. The amount of the cellulase used is preferably 0.01 to 1.0% by weight based on the mixture of rapeseed meal and water, etc. It is more preferable to add 0.05 to 1.0% by weight, and even more preferable to add 0.1 to 1.0% by weight. In addition, the enzyme used in the saccharification treatment may be any enzyme capable of decomposing fibrous material, and may be a commercially available product, a culture solution obtained by culturing a filamentous fungus or a further purified product thereof, or the filamentous fungus itself.

(Alcoholic fermentation process)
The alcohol fermentation treatment in the present invention refers to a process in which a microorganism is inoculated into a mixture of rapeseed meal and water, etc., or a mixture of rapeseed meal and water, etc., which has been subjected to saccharification treatment, and ethanol is produced by the microorganism. As for the microorganism to be used, yeasts such as Saccharomyces cerevisiae, Shizosaccharomyces pombe, and Pichia stipitis can be used, and other than yeasts, any microorganism capable of alcohol fermentation, including genetically modified ones, can be used, such as Zymomonas mobilis, which is a bacterium capable of alcohol fermentation. The microorganism may be one that has been preserved in a slant or frozen state, but when S. cerevisiae is used, commercially available baker's yeast may be used. When using a microorganism that has been preserved in a slant or frozen state, it is preferable to pre-culture it in a liquid medium before use and use the pre-culture liquid, etc. The liquid medium used for pre-culture may be one that is 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 in the alcohol fermentation treatment, and yeast that can be used in solid fermentation treatment is particularly preferred. The amount of yeast used is preferably such that a yeast suspension prepared so that the absorbance (hereinafter referred to as OD) when measured at 660 nm is 0.2 is added so that the amount of yeast present in the total reaction system (yeast (suspension) + meal + added water) is OD = 0.05 to 0.15. It is more preferable to add the yeast so that the OD is 0.05 to 0.1.

(When done in the same process or in separate processes)
In the case where saccharification and alcohol fermentation are carried out in the same process by adding enzymes and microorganisms to a mixture of rapeseed meal and water, etc. (referred to as "parallel fermentation"), the treatment temperature is preferably 25 to 45°C, more preferably 27 to 43°C, and even more preferably 30 to 40°C. In addition, when saccharification and fermentation are carried out in separate processes, 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, and even more preferably 25 to 35°C.
The total treatment time is preferably from 4 hours to 336 hours, more preferably from 24 to 72 hours, even more preferably from 24 to 72 hours, and most preferably from 24 to 72 hours.

(After solid-liquid separation, drying and distillation)
Rapeseed meal is mixed with water, and the mixture is subjected to saccharification and alcohol fermentation, followed by solid-liquid separation as necessary and drying. The product obtained as a result is called high-protein rapeseed meal. Drying may be performed by heat drying, vacuum drying, freeze drying, spray drying, or any other method capable of evaporating water and ethanol from the mixture that has been subjected to saccharification and/or alcohol fermentation. If ethanol is recovered by steam distillation or the like before drying, the recovered ethanol can be used for industrial purposes or as fuel.
In the present invention, the amount of ethanol produced per 100 g of dry weight of rapeseed meal is preferably 3.5 g or more, more preferably 3.8 g or more, and even more preferably 4.0 g or more.
The saccharification and alcohol fermentation are preferably carried out by solid fermentation, which is preferable because it does not require the above-mentioned solid-liquid separation and does not produce waste liquid.

(High protein rapeseed meal)
The high-protein rapeseed meal in the present invention is a high-protein rapeseed meal obtained by subjecting rapeseed meal to saccharification and alcohol fermentation, and contains 44 to 65 mass % protein, preferably 47 to 65 mass %, in terms of dry weight.
The protein content referred to here means the value obtained by multiplying the total nitrogen content determined by the Kjeldahl method by 6.25.
Formula feed using the high-protein rapeseed meal produced by the production method of the present invention does not pose any concerns about adverse effects on the growth or health of chicks, and can be used as feed without any problems.

(feed)
The high-protein rapeseed meal obtained by the production method of the present invention can be mixed with feed ingredients or feed additives containing sugars, proteins, amino acids, fiber, minerals, oils, antibacterial components, etc. and used as feed.
Specific examples of feed ingredients and feed additives include corn, sorghum, corn gluten field, corn starch, rice bran, soybean meal, ordinary rapeseed meal, bran, oats, milk casein, whey, fish meal, various vitamins, vitamin mixes, mineral mixes, amino acid preparations, calcium carbonate, monocalcium phosphate, vegetable fats and oils, animal fats and oils, and salt.
When the total amount of the feed is taken as 100% by mass, the content of the high protein rapeseed meal in the feed is preferably 0.1 to 30% by mass, more preferably 0.5 to 20% by mass, and even more preferably 0.5 to 10% by mass.
A feed containing high-protein rapeseed meal can be produced by mixing the high-protein rapeseed meal produced by the production method of the present invention with the above-mentioned various feed ingredients and feed additives. Mixing can be performed using a mixer such as a ribbon mixer, a V-type mixer, or a W-type mixer.

The following examples are provided to more specifically explain the present invention, but the present invention is not limited to these.

Example 1: Solid-state fermentation of rapeseed meal to ethanol by cellulose saccharification (small-scale test)
50 g of rapeseed meal (trade name: rapeseed oil cake, manufactured by Nisshin Oillio Group Co., Ltd., hereinafter the same) (moisture content 13.3% by mass) was added with 50 g of water to prepare 100 g of a mixture of rapeseed meal and water. A suspension of yeast (Saccharomyces cerevisiae) was added so that the total reaction system had an OD of 0.1 without autoclave sterilization. In addition, 0.05% by mass of each of the fibrous decomposition enzymes (cellulases) shown in Table 1 was added, and solid fermentation was performed at 35° C. for one week with the container lid closed. As the yeast (Saccharomyces cerevisiae), shochu yeast A30 (hereinafter referred to as "Saccharomyces cerevisiae A30") provided by the Laboratory of Brewing Microbiology, Department of Brewing Science, Tokyo University of Agriculture was used.
After one week, the solid portion was collected, its weight was measured, and the ethanol concentration in the solid portion was measured. The ethanol concentration was measured by adding the same amount of water to the solid portion, centrifuging at 1,000 rpm for 10 minutes, and collecting the supernatant. The mixture was then centrifuged at 15,000 rpm for 10 minutes, collecting the supernatant, diluting it four times with water, and measuring the ethanol concentration with a simple alcohol analyzer (product name: Alcomate, manufactured by Woodson Co., Ltd.). The results are shown in Table 2.

In Table 2, sufficient ethanol concentration was detected only when cellulase F was used as the fibrous enzyme. In other words, ethanol was produced so that 4.9 g of ethanol was produced per 100 g of dry weight of rapeseed meal. Cellulase F is an endoglucanase produced by microorganisms of the genus Acremonium, and it is considered that the high β-glucosidase activity influenced such good results. It is presumed that other fibrous enzymes were hardly able to decompose the fiber of rapeseed meal.
Therefore, the following experiment was conducted by focusing on cellulase F, which is produced by Acremonium microorganisms, as a fibrous enzyme.

Example 2: Evaluation of solid-state fermentation of rapeseed meal to ethanol in a medium-scale study
2500g of water was added to 2500g of rapeseed meal, the same as in Example 1, to prepare 5000g of a mixture of rapeseed meal and water. A suspension of yeast (Saccharomyces cerevisiae A30) was added so that the total reaction system had an OD of 0.1 without autoclave sterilization. In addition, 0.1% by mass of cellulase F shown in Table 2 was added, and solid fermentation was carried out at 35°C for 13 days with the container lid closed. An 8-liter resin case was used as the container.
The amount of ethanol produced per 100 g of dry weight of rapeseed meal in the medium-scale test is shown in FIG. 1, and the changes in each component of rapeseed meal (protein, total carbohydrate, cellulose) and the protein/carbohydrate ratio in the medium-scale test are shown in FIG.
Protein was measured by the Kjeldahl method, crude fat by the Soxhlet method, moisture by normal pressure heating and drying method, and crude ash by direct ashing method (weight). Carbohydrates were calculated by subtracting the total value of these from 100.

As is clear from Figure 1, ethanol was produced so that the amount of ethanol produced per 100 g of dry weight of rapeseed meal was 4.3 to 4.6 g. This result is not very different from the small-scale test, but it means that about twice the amount of ethanol was produced compared to the conventional method (about 2.7 g: see Table 1 of Patent Document 2). In other words, it was found that the use of cellulase F improves ethanol production ability.
2, the protein content of the rapeseed meal before and after fermentation increased from 44.8% to 47.1%. This indicates that the protein content of the rapeseed meal was relatively increased by converting the fiber content of the rapeseed meal into ethanol by fermentation.
Meanwhile, the total carbohydrate content before and after fermentation decreased from 44.3% to 40.0%, and the cellulose content before and after fermentation decreased from 11.4% to 9.4%, indicating that the fiber content of the rapeseed meal was converted into ethanol by fermentation.
In addition, the protein/carbohydrate ratio on the 10th day of fermentation was improved from 1.01 to 1.20 compared to that before fermentation. This indicates that the use of Cellulase F reduced the fiber content, resulting in an improved protein/carbohydrate ratio.

Example 3: Solid-state fermentation of rapeseed meal to produce ethanol using high concentrations of fibrous enzymes (small-scale test)
10 g of rapeseed meal (trade name: rapeseed oil cake, manufactured by Nisshin Oillio Group Co., Ltd., the same applies below) (moisture content 13.3% by mass) was added with 10 g of water to prepare 20 g of a mixture of rapeseed meal and water. A suspension of yeast (Saccharomyces cerevisiae A30) was added so that the total reaction system had an OD of 0.1 without autoclave sterilization. In addition, 1.0% by mass (10 times the amount of Example 1) of each of the cellulases shown in Table 3 was added, and solid fermentation was carried out at 30° C. for 3 days with the lid of the container closed.
The ethanol concentration was measured in the same manner as in Example 1. The results are shown in FIG.

According to Figure 3, when various fibrous enzymes were added at a high concentration (1.0% by mass), cellulase F produced by Acremonium microorganisms produced 6.5 g of ethanol per 100 g of dry rapeseed meal, indicating that the ethanol yield was higher than that of other cellulases. However, even when the fibrous enzyme concentration was changed from 0.1% by mass to 1.0% by mass, which is a tenfold increase, the ethanol yield only increased by about 1.3 times (note that at 0.1% by mass, the amount of ethanol produced per 100 g of dry rapeseed meal was 5.2 g), confirming that an amount of enzyme used of 0.1% by mass is sufficient.

Example 4: Optimum temperature for saccharification when saccharification and alcohol fermentation are performed separately (small-scale test)
10 g of rapeseed meal (trade name: rapeseed oil cake, manufactured by Nisshin Oillio Group Co., Ltd., the same applies below) (moisture content 13.3% by mass) was added with 10 g of water to prepare 20 g of a mixture of rapeseed meal and water. Without autoclave sterilization, 0.1% by mass of each of the fibrous enzymes shown in Table 4 was added and saccharification treatment was performed at 45°C or 50°C for 3 days. Thereafter, a suspension of yeast (Saccharomyces cerevisiae A30) was added so that the total reaction system had an OD of 0.1, and solid fermentation was performed at 40°C for 3 days with the container lid closed. The ethanol concentration was measured in the same manner as in Example 1 above. The results are shown in FIG. 4.

According to Figure 4, when various fibrous decomposition enzymes were added at 0.1% by mass, the fermentation efficiency of cellulase produced by Acremonium microorganisms was higher when saccharification was performed at 50°C than when saccharification was performed at 45°C. Therefore, it was found that when using cellulase produced by Acremonium microorganisms, saccharification can be performed at 50°C to prevent insufficient yeast fermentation due to contamination by various bacteria, etc.

Example 5: Optimum temperature for alcoholic fermentation when saccharification and alcoholic fermentation are performed separately (small-scale test)
10 g of rapeseed meal (trade name: rapeseed oil cake, manufactured by Nisshin Oillio Group Co., Ltd., the same applies below) (moisture content 13.3% by mass) was added with 10 g of water to prepare 20 g of a mixture of rapeseed meal and water. In Example 5, saccharification treatment and alcoholic fermentation were performed separately. Without autoclave sterilization, 0.1% by mass of each of the fibrous enzymes shown in Table 5 was added and saccharification treatment was performed at 50°C for 3 days. Then, a suspension of yeast (Saccharomyces cerevisiae A30) was added so that the total reaction system had an OD of 0.1, and solid fermentation was performed at 30°C or 40°C for 3 days with the container lid closed. The ethanol concentration was measured in the same manner as in Example 1 above. The results are shown in FIG. 5.

According to Figure 5, when various fibrous enzymes were used for saccharification at 50°C, the ethanol yield could be increased by lowering the temperature of the alcoholic fermentation process to 30°C rather than 40°C. It was also found that, among cellulases produced by Acremonium microorganisms, Cellulase KM gave a higher ethanol yield than Acremonium Cellulase F. However, when saccharification and alcoholic fermentation were performed separately, a maximum of only 4.4 g of ethanol was produced per 100 g of dry weight of rapeseed meal, so the ethanol yield was not much higher than when they were performed by simultaneous fermentation. Therefore, it was confirmed that simultaneous fermentation is more preferable than separate saccharification and alcoholic fermentation.

Example 6: Solid-state fermentation of rapeseed meal to ethanol using various yeasts (small-scale test)
10 g of rapeseed meal (trade name: rapeseed oil cake, manufactured by Nisshin Oillio Group Co., Ltd., the same below) (moisture content 13.3% by mass) was added with 10 g of water to prepare 20 g of a mixture of rapeseed meal and water. Without autoclave sterilization, various yeast suspensions shown in Table 5 were added so that the total reaction system had an OD of 0.1, and 0.1% by mass of Acremonium Cellulase F was added, and solid fermentation was performed at 37°C for 3 days with the container lid closed. Thereafter, the ethanol concentration was measured in the same manner as in Example 3 above. The results are shown in Figure 6.

Figure 6 shows that when various types of yeast were used, similar amounts of ethanol were produced when cellulase produced by microorganisms of the genus Acremonium was used, as was the case when Saccharomyces cerevisiae A30 was used. Therefore, any type of yeast can be used, and it seems that the amount of ethanol produced does not vary significantly depending on the type of yeast.

<Feeding test of compound feed containing high protein rapeseed meal>
- Feed safety confirmation test Fermented rapeseed meal was produced in the same manner as in Example 2, except that 4000 g of water was added to 4000 g of rapeseed meal to obtain a mixture of 8000 g of rapeseed meal and water. The obtained fermented rapeseed meal was subjected to reduced pressure heat treatment (72°C, -0.1 MPa) for 8 hours. Thereafter, the dried fermented rapeseed meal was pulverized to a size suitable for feed ingredients using a pulverizer (Osaka Chemical Co., Ltd., device "Absolute mill ABS-W") to produce high protein rapeseed meal (protein content: 49.4% (dry matter equivalent)).
The safety of compound feed containing the obtained high-protein rapeseed meal was confirmed in accordance with the "Chicken Growth Test" in the "Establishment of Feed Safety Evaluation Standards and Evaluation Procedures (Notice No. 20-597 dated May 19, 2008, by the Director-General of the Food Safety and Consumer Affairs Bureau, Ministry of Agriculture, Forestry and Fisheries)," and it was confirmed that the high-protein rapeseed meal of the present invention can be used in feed without any problems.
1) Test chicks The basic feed shown in Table 7 (rapeseed meal (product name: rapeseed oil cake, manufactured by Nisshin Oillio Group, Ltd.) 20.00% by mass) was given to each chick at the time of feeding, at a rate of 10 g for three days, and from the fourth day onwards, 3.5 g per chick per day was given to each chick. From these chicks, 8-day-old laying hens (Julia Lite) male chicks weighing 42 to 46 g were selected for the test.
The basic feed used was designed so that the contents of metabolizable energy, general components such as protein, various minerals, amino acids, etc., would satisfy the nutrient requirements of egg-laying chicks as specified in the Japanese Feeding Standards for Poultry (2011 edition).
2) Setting up test plots Two test plots were set up, where the animals were fed compound feed 1 (containing 10.00 mass% rapeseed meal (product name: rapeseed oil cake, manufactured by Nisshin Oillio Group Ltd.) and 10.00 mass% high-protein rapeseed meal) shown in Table 7, and compound feed 2 (containing 20.00 mass% high-protein rapeseed meal).
Formula feeds 1 and 2 were designed so that the contents of metabolizable energy, general components such as protein, various minerals, amino acids, etc., would satisfy the nutrient requirements of egg-laying chicks as specified in the Japan Feeding Standards for Poultry (2011 edition).
The contents of each component in the B vitamin premix used in the compound feed were thiamine nitrate 2.0 g/kg, riboflavin 10.0 g/kg, pyridoxine hydrochloride 2.0 g/kg, nicotinamide 2.0 g/kg, D-calcium pantothenate 4.35 g/kg, choline chloride 138.0 g/kg, folic acid 1.0 g/kg, and cyanocobalamin 10 mg/kg.
The content of each component in the vitamin ADE premix is 10,000 IU/kg of vitamin A oil, 2,000 IU/kg of vitamin _D3 oil, and 20 mg/kg of dl-α-tocopherol acetate.
The content of each component in the mineral premix is as follows: Mn 80 g/kg, Zn 50 g/kg, Fe 6 g/kg, I 1 g/kg, and Cu 0.6 g/kg.

3) Establishment of test groups and rearing management The test chicks were divided into six groups of six chicks each so that the weight distribution was approximately equal, and each group was divided into three replicate groups and reared for six days.
The chicks were reared in groups in electrically heated brooders and given various compound feeds and drinking water ad libitum. To prevent the influence of environmental conditions, the housing locations of each group were moved every day.
4) Weight gain, feed intake and feed conversion rate of test chicks As reference data, the weight of each test chick was measured at the start and end of the test, and the weight gain (g/bird) during the test period was calculated.
As reference data, the feed intake during the test period was measured for each group, and the feed intake per bird (g/bird) and feed conversion rate were calculated.
The results of these measurements (mean values ± standard deviations for each of the three groups) are shown in Table 8.

As a result of the feeding test, no abnormalities were observed in the health of any of the individuals in either test group. This confirmed that there is no risk of adverse effects on the growth or health of chicks from compound feed containing the high-protein rapeseed meal of the present invention, and that it can be used as feed without any problems.

Claims (8)

A method for producing high-protein rapeseed meal, which comprises mixing rapeseed meal with water, subjecting the resulting mixture to saccharification and alcoholic fermentation, adding a fibrous decomposing enzyme for the saccharification, and adding a microorganism for the alcoholic fermentation, characterized in that cellulase produced by Acremonium cellulolyticus is used as the fibrous decomposing enzyme , and the mixture of rapeseed meal and water is not sterilized before the saccharification and/or alcoholic fermentation are performed . The method according to claim 1, characterized in that the amount of ethanol produced per 100 g of dry weight of rapeseed meal in the alcohol fermentation treatment is 3.5 g or more. The method according to claim 1 or 2, characterized in that 40 to 60 parts by weight of the rapeseed meal is mixed with 60 to 40 parts by weight of water. The method according to any one of claims 1 to 3, characterized in that the saccharification process and the alcohol fermentation process are carried out in the same process. The method of claim 4, characterized in that the same process is carried out at 25 to 45°C. The method according to any one of claims 1 to 3, characterized in that the saccharification process and the alcohol fermentation process are carried out in separate steps. The method according to claim 6, characterized in that the saccharification process is carried out at 40 to 60°C, and the alcohol fermentation process is carried out at 20 to 40°C. A feed containing high-protein rapeseed meal produced by the method according to any one of claims 1 to 7 .