Method for producing high protein rapeseed meal

Abstract

To provide a high protein rapeseed meal producing method, in which industrially valuable ethanol is produced, and the high protein rapeseed meal excellent in nutritional property is produced simultaneously, to increase ethanol productivity and improve the protein ratio.SOLUTION: In a high protein rapeseed meal producing method, a rapeseed meal and water are mixed; saccharification treatment and alcohol fermentation treatment are performed on the obtained mixture; a fiber-degrading enzyme is added for the saccharification treatment; and microorganisms are added for the alcohol fermentation treatment. In the high protein rapeseed meal producing method, cellulase produced by microorganisms belonging to the genus Acremonium is used as the fiber-degrading enzyme.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a method for producing a high protein rapeseed meal. [Background technology]

[0002] The residue left after removing oil from oilseeds such as rapeseed and soybeans is mainly used as a protein source for animal feed. However, rapeseed meal, which is the residue left after the rapeseed is pressed, has the disadvantage that it has a lower protein content than soybean meal and a higher fiber content, resulting in a lower energy value for livestock.

[0003] As a method for solving such a drawback, several methods for increasing the protein content in rapeseed meal have been proposed. For example, a method has been disclosed in which rapeseed meal is fractionated with a sieve into a fraction with a high protein content and a fraction with a low protein content, but the low protein fraction has low feed value and its range of use is limited (Patent Document 1). Also, a method has been disclosed in which rapeseed is peeled, then oiled and rapeseed meal is recovered, but this method of peeling rapeseed with small particle size is expensive and not practical (Non-Patent Document 1). Furthermore, a method for producing high-protein, low-glucosinolate rapeseed meal has been disclosed in which rapeseed meal is mixed with water, and the mixture is subjected to saccharification and / or alcohol fermentation, followed by solid-liquid separation and drying (Patent Document 2), but further improvements in ethanol production capacity and the like have been desired.

[0004] [Patent Document 1] Patent No. 3970917 [Patent Document 2] Patent No. 5601763 [Non-Patent Document 1] W. Grala, et al., Ileal 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) DISCLOSURE OF THEINVENTION [Problem to be solved by the invention]

[0005] The object of the present invention is to improve the ethanol productivity in a method for producing high-protein rapeseed meal, which is capable of producing industrially valuable ethanol and simultaneously producing high-protein rapeseed meal with excellent nutritional properties. [Means for solving the problem]

[0006] In order to solve the above problems, the inventors conducted extensive research and discovered that ethanol production capacity can be increased by using cellulase produced by Acremonium microorganisms as a fiber-degrading enzyme for use in saccharification treatment, thereby completing the present invention.

[0007] Specifically, the present invention may include the following aspects. [1] A method for producing high-protein rapeseed meal, comprising mixing rapeseed meal with water, subjecting the mixture to saccharification and alcohol fermentation, adding a fibrous enzyme for the saccharification, and adding a microorganism for the alcohol fermentation, comprising: Cellulases produced by Acremonium cellulolyticus Using The mixture of rapeseed meal and water is not sterilized before the saccharification treatment and / or alcohol fermentation treatment. , A method for producing high protein rapeseed meal. [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 and 60 to 40 parts by weight of water are mixed. [4] The production method described in 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 process is carried out at 25 to 45°C. [6] The production method described in 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 production method described in [6], characterized in that 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 〕 the above 〔1〕~〔7〕 A feed containing high-protein rapeseed meal produced by the production method described in any one of the above. Effect of the Invention

[0008] 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 is used for various purposes as bioethanol. Furthermore, whether the saccharification treatment and the alcohol fermentation treatment 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 treatment (autoclave treatment, etc.) that was previously required, and therefore high-protein rapeseed meal can be produced in a simpler manner than before. Furthermore, when the solid fermentation method is used in the present invention, no waste liquid is produced, making it possible to provide an environmentally friendly method for producing high-protein rapeseed meal. [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 shows ethanol production per 100 g dry weight of rapeseed meal (medium-scale study) (days 7 to 13). [Diagram 2]This is a graph showing the changes in rapeseed meal components (protein, total carbohydrate, cellulose) and protein / carbohydrate ratio (medium-scale test) (before fermentation to 10th or 13th day). [Diagram 3] FIG. 1 shows ethanol production per 100 g dry weight of rapeseed meal using various fibrous enzymes at high concentrations (small scale study). [Figure 4] FIG. 1 shows the optimum temperature for saccharification (small-scale test) when saccharification and fermentation are separated. [Diagram 5] FIG. 1 shows the optimum temperature for alcohol fermentation treatment (small-scale test) when saccharification treatment and fermentation treatment are separated. [Figure 6] FIG. 1 shows ethanol production per 100 g dry weight of rapeseed meal using different yeasts (small scale study). BEST MODE FOR CARRYING OUT THEINVENTION

[0010] (rapeseed meal) Rapeseed meal is a rapeseed meal produced by extracting rapeseeds 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. It contains 8 to 15% moisture by mass.

[0011] (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 liquid fermentation method, and almost no waste liquid is discharged not only during the fermentation period but also after the fermentation is completed.

[0012] Additives such as minerals, sugars, amino acids, medium components, enzymes, and microorganisms 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, and iron. Specific examples of sugars include sucrose, glucose, maltose, and oligosaccharides. Specific examples of amino acids include glutamic acid and lysine, and may 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, and peptone. Specific examples of the enzyme for the saccharification treatment include cellulase, hemicellulase, glucosidase, and a mixture thereof. In the saccharification treatment of the present invention, as one type of the cellulase (fibrous decomposition enzyme), cellulase produced by a microorganism of the genus Acremonium, particularly Acremonium cellulolyticus ), Cellulase F produced by the enzyme (product name: Meiji Acremonium Cellulase F, manufactured by Meiji Seika Pharma Co., Ltd.), Acremonium cellulolyticus In the present invention, Cellulase KM produced by Acremonium Cellulase KM (product name: Acremonium Cellulase KM, manufactured by Kyowa Kasei Co., Ltd.) is used. Acremonium cellulolyticus As described below, it has been discovered for the first time in the present invention that the use of cellulases produced by these Acremonium microorganisms improves ethanol production ability. 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.

[0013] (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.

[0014] (Saccharification) The saccharification treatment in the present invention refers to adding a fibrous decomposition enzyme to a mixture of rapeseed meal, 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. Also, 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.

[0015] (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 preferable. The amount of the 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.

[0016] (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 (called "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. 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 4 to 336 hours, more preferably 24 to 72 hours, further preferably 24 to 72 hours, and especially preferably 24 to 72 hours.

[0017] (After solid-liquid separation, drying and distillation) Rapeseed meal is mixed with water and subjected to saccharification and alcohol fermentation, and then solid-liquid separation is performed as necessary and dried. The resultant product is called high-protein rapeseed meal. Drying can be performed by any method capable of evaporating water and ethanol from the mixture that has been subjected to saccharification and / or alcohol fermentation, such as dry heat drying, vacuum drying, freeze drying, spray drying, etc. 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.

[0018] (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 treatment, and contains 44 to 65 mass % protein in dry weight terms, and preferably 47 to 65 mass % protein. The protein content referred to here refers to the total nitrogen content determined by the Kjeldahl method multiplied 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.

[0019] (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.

[0020] In order to more specifically explain the present invention, examples are shown below, but the present invention is not limited thereto.

[0021] <Example 1: Solid-state fermentation of rapeseed meal to ethanol by cellulose saccharification (small-scale test)> 50 g of rapeseed meal (product name: rapeseed oil cake, manufactured by Nisshin Oillio Group Co., Ltd., the same below) (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 carried out 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 and weighed, and then 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. It was then centrifuged at 15,000 rpm for 10 minutes, and the supernatant was collected and diluted four times with water, and the ethanol concentration was measured using a simple alcohol analyzer (product name: Alcomate, manufactured by Woodson Co., Ltd.). The results are shown in Table 2.

[0022] [Table 1]

[0023] [Table 2]

[0024] In Table 2, sufficient ethanol concentration was detected only when cellulase F was used as the fibrous decomposition enzyme. In other words, ethanol was produced so that 4.9 g of ethanol was obtained per 100 g of dry weight of rapeseed meal. Cellulase F is an endoglucanase produced by microorganisms of the genus Acremonium, and it is believed that the high β-glucosidase activity influenced such good results. It is presumed that other fibrous decomposition enzymes were barely able to decompose the fiber in rapeseed meal. Therefore, the following experiment was conducted by focusing on cellulase F, which is produced by Acremonium microorganisms, as a fibrous enzyme.

[0025] 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. Figure 1 shows the amount of ethanol produced per 100 g of dry weight of rapeseed meal in the medium-scale test, and Figure 2 shows the changes in each component of rapeseed meal (protein, total carbohydrate, cellulose) and the protein / carbohydrate ratio in the medium-scale test. Protein was measured by the Kjeldahl method, crude fat by the Soxhlet method, moisture by the normal pressure heating drying method, and crude ash by the direct ashing method (by weight). Carbohydrates were calculated by subtracting the total value of these from 100.

[0026] As is clear from FIG. 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. In addition, as is clear from Figure 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 in the rapeseed meal increased relatively because the fiber in the rapeseed meal was converted 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.

[0027] 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 (product 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. 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 container lid closed. The ethanol concentration was measured in the same manner as in Example 1. The results are shown in FIG.

[0028] [Table 3]

[0029] 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 weight 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 when the concentration was 0.1% by mass, the amount of ethanol produced per 100 g of dry weight rapeseed meal was 5.2 g), indicating that the amount of enzyme used was sufficient at 0.1% by mass.

[0030] <Example 4: Optimum temperature for saccharification treatment when saccharification treatment and alcohol fermentation treatment are performed separately (small-scale test)> 10 g of rapeseed meal (product 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, 0.1% by mass of each of the fibrous enzymes shown in Table 4 was added and saccharification treatment was carried out at 45°C or 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 carried out 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.

[0031] [Table 4]

[0032] 4, when various fibrous 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.

[0033] <Example 5: Optimum temperature for alcohol fermentation when saccharification and alcohol fermentation are performed separately (small-scale test)> 10 g of rapeseed meal (product 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. 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 Figure 5.

[0034] [Table 5]

[0035] According to FIG. 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, only 4.4 g of ethanol was produced per 100 g of dry weight of rapeseed meal at most, so the ethanol yield was not much higher than when they were performed by simultaneous fermentation. Therefore, it was also confirmed that simultaneous fermentation is more preferable than separate saccharification and alcoholic fermentation.

[0036] <Example 6: Solid-state fermentation of rapeseed meal to ethanol using various yeasts (small-scale test)> 10 g of rapeseed meal (product name: rapeseed oil cake, manufactured by Nisshin Oillio Group, 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, 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. With the lid of the container closed, solid fermentation was carried out at 37°C for 3 days. Thereafter, the ethanol concentration was measured in the same manner as in Example 3 above. The results are shown in Figure 6.

[0037] [Table 6]

[0038] According to Fig. 6, when various kinds of yeast were used, similarly to the case of using Saccharomyces cerevisiae A30, when cellulase produced by microorganisms of the genus Acremonium was used, all of them produced a sufficient amount of ethanol to the same extent. Therefore, any kind of yeast can be used, and it seems that the amount of ethanol produced does not vary greatly depending on the type of yeast.

[0039] <Feeding test of compound feed containing high protein rapeseed meal> Feed safety verification 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 a reduced pressure heat treatment (72°C, -0.1 MPa) for 8 hours. The dried fermented rapeseed meal was then crushed into a size suitable for feed ingredients using a crusher (Osaka Chemical Co., Ltd., "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 and Agriculture Food Safety 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 fed to each chicken at 10 g for three days at feeding time, and from the fourth day onwards, 3.5 g per chicken per day was fed to each chicken. From these chickens, 8-day-old laying hens (Julia Light) 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 Japan Feeding Standards for Poultry (2011 edition). 2) Setting up test areas Two test groups were set up, each 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 content of each component in the B vitamin premix used in the compound feed was 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 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 80g / kg, Zn 50g / kg, Fe 6g / kg, I 1g / kg, and Cu 0.6g / kg.

[0040] [Table 7]

[0041] 3) Setting up test areas and feeding management The test chicks were divided into six groups of six chicks each with approximately equal weight distribution, and each group was divided into three replicates 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 measurement results (mean value ± standard deviation for each of the three groups) are shown in Table 8.

[0042] [Table 8]

[0043] As a result of the feeding test, no abnormalities were observed in the health of any of the individuals in both test groups. This confirmed that there is no concern that the compound feed containing the high protein rapeseed meal of the present invention will have a negative effect on the growth or health of chicks, and that it can be used as a feed without any problems.

Claims

1. 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.

2. The method according to claim 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 according to claim 1 or 2, characterized in that 40 to 60 parts by weight of the rapeseed meal and 60 to 40 parts by weight of water are mixed.

4. The method according to claim 1 , wherein the saccharification treatment and the alcoholic fermentation treatment are carried out in the same step.

5. The method according to claim 4, wherein the same step is carried out at 25 to 45°C.

6. The method according to any one of claims 1 to 3, wherein the saccharification treatment and the alcoholic fermentation treatment are carried out in separate steps.

7. The method according to claim 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 method according to any one of claims 1 to 7.

Images