JP4999357B2 - Method for producing fermented feed with soy sauce cake - Google Patents

Description

  The present invention relates to a method for producing a fermented feed containing soy sauce cake. More specifically, the present invention relates to a soy sauce cake produced by the action of microorganisms belonging to the species of Neurospora sitophila or Rhizopus oligosporus having phytase producing ability. The present invention relates to a method for producing a fermented feed containing soy sauce cake that deodorizes a specific odor and also functions as a supply source of phytase.

  Soy sauce is consumed in large quantities as an essential item indispensable for the Japanese diet, and the amount of soy sauce cake produced as a by-product reaches several hundred thousand tons per year. An effective method for treating this soy sauce cake is one of the important issues for those skilled in the art.

  Most soy sauce cakes at present are treated as industrial waste by incineration and the like, and only a very small part is effectively used as livestock feed or fertilizer for agricultural products. The reason is that because soy sauce cake contains a large amount of salt, the amount of feed and fertilizer is limited to a very small amount (less than 5%, usually about 1-2%), soy sauce cake has a strong odor, Even if the mixed feed is produced, the odor derived from the soy sauce cake remains strong, and the value of the feed is reduced. In addition, the odor of this soy sauce cake is not shunned by workers, but the use of soy sauce cake is also due to the fact that the odor derived from the soy sauce cake remains in the equipment used when preparing mixed feed etc. The range was limited.

  Also, incineration has many problems such as the need to use a special incinerator to prevent metal corrosion and dioxin generation due to salt. Furthermore, the Food Recycling Law has been enacted, and the reduction and recycling of industrial waste at the soy sauce factory has become a problem. From the viewpoint of effective use of waste, it is desired to use it for feed and fertilizer.

  Various methods have been devised in order to solve these problems, but none of them has a sufficient effect. All of these methods can solve some of the problems of soy sauce cake (salt and odor of soy sauce cake), but at present, soy sauce cake feed and fertilizer are high in nutritional value and easily available at low cost. It is thought that the reason why it is not practical is that the value of the product is low and is not profitable. Therefore, it can be said that, in order to effectively use soy sauce cake, it is very important to improve the added value of the processed product as well as to reduce salt and deodorization.

  Meanwhile, eutrophication of rivers and lakes by phosphorus contained in livestock manure is one of the problems in the livestock field, which is a field of use of the dominant soy sauce cake. This is because monogastric animals such as poultry and pigs do not secrete phytase in the digestive tract, and only about 30% of the phytin contained in grains can be used. The cause is that it is used according to the demand. Nowadays, phytase (an enzyme that generates phosphate and inositol from phytin) is constantly added to poultry and swine feeds to reduce the addition of mineral phosphorus.

  The amount of phytase added to general feed is said to be 200 to 500 units / kg for chickens and 300 to 1000 units / kg for pigs. Here, one unit of phytase is defined as the amount of enzyme that produces 1 micromole of inorganic phosphate per minute at 37 ° C.

  Currently, the phytase agent used for the above-mentioned purpose is a phytase mainly derived from Aspergillus genus using a gene recombination technique. However, nowadays, where food safety and security are required, the use of an enzyme preparation using genetic recombination technology may raise public anxiety.

  Non-genetically modified phytase-producing bacteria include Aspergillus genus (Aspergillus phycum, Aspergillus fumigatus, Aspergillus niger, etc.), Penicillium genus, Rhizopus genus, Neurospora genus yeast, Saccharomyces yeast, Bacillus, etc. Bacteria are mentioned. However, from the viewpoint of effective use of soy sauce cake, fermentation by microorganisms that can be eaten as they are is preferable. Not only harmful bacteria such as Aspergillus phycumum and Aspergillus fumigatus, but also Aokabi (such as Penicillium) used in shochu (Aspergillus niger, Aspergillus kawachi, etc.) and blue cheese are eaten by removing mold cells However, although there is a dietary habit, even the feed that is added as it is does not guarantee its safety.

  From such a viewpoint, phytase-producing bacteria that can withstand edible use include Aspergillus oryzae used for miso, Rhizopus / oligosporus used for tempeh, and Rhizospora used for onchome. The present inventors have discovered that phytase is accumulated in a medium by solid-culturing fungi of Neurospora and Rhizopus in a medium such as wheat bran (Japanese Patent No. 2287737).

  Therefore, the present invention provides a method for producing a fermented feed containing soy sauce cake in which the odor of the soy sauce cake is deodorized by a safe means and helps to supply phytase.

  As a result of various studies to solve the above-mentioned problems, the present inventors have confirmed that it is safe even if directly mixed with food, and have the ability to produce phytase-producing Neurospora sitophila species or Rhizopus.・ By culturing microorganisms belonging to Rhizopus oligosporus species in a solid medium containing soy sauce cake, the odor of soy sauce cake can be deodorized and an amount of phytase sufficient to help supply phytase. The present invention was completed by finding out the accumulation.

  Accordingly, the present invention provides a mixture of 90% by weight or less of soy sauce cake and 10% by weight or more of solid feed raw material with respect to the total weight of solids and 80 to 150% with respect to the weight of the mixture. Culturing a microorganism belonging to the species of Neurospora sitophila or Rhizopus oligosporus having the ability to produce phytase in a solid medium comprising 1% water and accumulating 2 units / g or more of phytase. Provided is a method for producing a fermented feed containing soy sauce cake.

  In the above method, from the viewpoint of the completeness of deodorization and the accumulated amount of phytase, the amount of soy sauce cake with respect to the total weight of the solid is preferably 70% by weight or less, and the amount of the solid feed raw material with respect to the total weight of the solid is 30. %, More preferably the amount of soy sauce cake with respect to the total weight of the solid is 60% by weight or less, the amount of the solid feed material with respect to the total weight of the solid is 40% or more, and more preferably The amount of soy sauce cake with respect to the weight is 50% by weight or less, and the amount of the solid feed raw material with respect to the total weight of the solid material is 50% or more.

Preferably, the accumulated amount of phytase is 4 units / g or more, more preferably 9 units / g or more, and further preferably 15 units / g or more.
As a preferred example of the above-mentioned Neurospora sitophila, the Neurospora sitophila NA38 strain (NITE) deposited at the Microorganism Deposit Center, National Institute of Technology and Evaluation (2-5-8, Kazusa Kamashi, Kisarazu City, Chiba Prefecture) AP-229) and Neurospora sitophila NA37 strain (NITE AP-230), and preferred examples of the Rhizopus oligosporus include Rhizopus oligosporus, Rhizopus oligosporus KT28 strain (NITE AP-232), and Rhizopus oligospora KT27 strain. (NITE AP-231).

The solid feed material is, for example, wheat bran, rice bran, tofu koji, oil koji, soybean koji, koji koji, corn gluten meal, raw ore that is by-produced during the production of soy sauce, It is a mixture of two or more.
In 1 aspect of this invention, fats and oils and soy sauce oil are further added.
The present invention further comprises a method for producing a mixed feed containing soy sauce cake, wherein the fermented feed containing soy sauce cake produced as described above is added to the feed in an amount of 10% by weight or less based on the total weight of the feed. I will provide a.

Microorganisms In the present invention, microorganisms belonging to the species of Neurospora sitophila or Rhizopus oligosporus are used as microorganisms having the ability to produce phytase and whose safety has been confirmed. These microorganisms have the ability to deodorize the smell of soy sauce cake regardless of the degree of phytase production ability. Therefore, the selection of microorganisms is performed mainly in consideration of the ability to produce phytase.

  Generally, the amount of phytase added to feed is said to be 200 to 500 units / kg for poultry feed and 300 to 1000 units / kg for pig feed. Therefore, considering that the soy sauce cake contains salt, the amount added to the feed is limited to less than 5%, the amount of phytase in the fermented feed containing soy sauce cake of the present invention is 20 units / g or more. It is desirable to be. However, it is not necessary to cover all of the phytase added to the feed with the fermented feed containing soy sauce cake of the present invention, and it only has to help the addition of phytase to the feed. Therefore, the amount of phytase in the fermented feed containing soy sauce cake of the present invention may be 2 units / g or more.

Accordingly, examples of microorganisms used in the present invention include known microorganisms known to have phytase producing ability, such as Neurospora sitophila NBRC 31635, Rhizopus oligosporus NBRC 8631, and the like. These can be obtained from NBRC (National Institute of Technology and Evaluation).
However, as described above, the microorganism of the present invention preferably has a phytase-producing ability as high as possible. For this purpose, it is preferable to obtain a phytase high-producing mutant. The mutant strain can be obtained according to a conventional method for obtaining the target mutant strain.

  The specific method is shown in Example 2. According to it, as for Neurospora sitophila, several high phytases such as Neurospora sitophila NA38 strain (NITE AP-229), Neurospora sitophila NA37 strain (NITE AP-230), and Neurospora sitophila NA33 strain are produced. Strains were obtained with relatively high efficiency (approximately 1 in 10,000). Therefore, according to the present invention, not only the mutant strains deposited above, but also equivalent mutant strains can be easily created and used. Note that the neurospora sitophila NA38 strain (NITE AP-229) and the neurospora sitophila NA37 strain (NITE AP-230) have a phytase producing ability of at least 20 units / g (Example 3).

  As for Rhizopus oligosporus, several high phytase producing strains such as Rhizopus oligosporus KT28 strain (NITE AP-232), Rhizopus oligosporus KT27 strain (NITE AP-231) and Rhizopus oligosporus KT26 are compared. High efficiency (about 1 in 10,000). Therefore, according to the present invention, not only the mutant strains deposited above, but also equivalent mutant strains can be easily created and used. In addition, Rhizopus-oligosporus KT28 strain (NITE AP-232) has a phytase production ability of at least 6 units / g or more (Example 3).

  In the present invention, one unit of phytase is reacted at 37 ° C. in 100 mM acetate buffer (pH 5.7) containing 5 mM sodium phytate solution to release 1 micromole of inorganic phosphate per minute. It is defined as the amount of enzyme to be made.

Soy sauce cake The soy sauce cake that can be used in the present invention is not limited to a special one. That is, soy sauce cake produced as a by-product by various methods such as a main brewing method, a new brewing method, an amino acid solution mixing method, and an enzyme treatment solution mixing method can be used as a raw material. These soy sauce cakes may be dried or not. In addition to normal soy sauce koji, it produces fish soy sauce such as water-insoluble fraction obtained by hydrolyzing koji in the presence of salt, water-insoluble fraction obtained when producing soy sauce-like seasoning liquid, and shochuru. Water-insoluble fractions obtained during production of various salt-containing seasonings such as water-insoluble fractions obtained at the time can also be used as a raw material for the method of the present invention. Therefore, “soy sauce cake” in the present specification includes all of the above.

The feed material to be mixed with the feed material is not particularly limited as long as it can be used as the solid medium of the microorganism of the present invention. For example, wheat bran, rice bran, tofu koji, oil koji, soybean koji, koji koji, corn gluten meal by-produced in the production of corn starch, raw or by-products produced during the production of soy sauce, etc. A wide range of unused resources can be used. Two or more of these may be used in combination.

Preparation of Medium In preparation of the medium, a predetermined amount of soy sauce cake and a predetermined amount of the above feed raw material are mixed, and this is added to the total weight (100%) of the soy sauce cake and the feed raw material. Add ~ 150 wt% water and mix the whole. Here, the amount of water of 80 to 150% means, for example, that 80 to 150 kg of water is added to a total weight of 100 kg of soy sauce cake and feed raw materials. If the amount of water is too small, the growth of microorganisms will be insufficient, and if the amount of water is too large, the ventilation in the solid medium will be poor and the growth of microorganisms will also be poor. Next, sterilization is performed. The sterilization may be, for example, steam sterilization (autoclave sterilization) at 121 ° C. (gauge pressure: 1 atm) for 15 to 30 minutes.
It is not necessary to prepare a strawberry as an inoculum for culture culture, and the inoculation of the microorganism can be completed simply by mixing the spore with the soy sauce candy-containing solid medium. The inoculated amount of spore may be about 1 × 10 ³ cells / g solid medium or more, preferably 1 × 10 ⁵ to 10 ⁶ cells / g solid medium. When the inoculation amount is smaller than the above range, the culture time is prolonged, or contamination with other microorganisms is not preferable. The culture temperature may be the normal growth range of Aspergillus oryzae, and temperature control is not particularly required, but a range of 25 to 40 ° C. is suitable as a favorable condition for germination of spore and growth of Aspergillus.

  Aeration during the culture is not particularly required, but it may be appropriately aerated to prevent the growth of the bacteria from being stopped and the bacteria from being killed due to lack of oxygen. The culture time varies depending on the water content, culture temperature, pH and the like, but usually a culture time of about 16 to 48 hours is appropriate. The method for adding soy sauce oil is not particularly limited, but it may be added to the medium from the beginning, but it is preferably added immediately before the end of fermentation in consideration of enzyme production. The completion of the culture and drying can be performed by heat drying at 30 to 70 ° C.

In the production of soy sauce with added soy sauce, not only soy sauce cake, but also soy sauce oil is produced as a by-product, and soy sauce has a soy odor and cannot be used as it is. However, if it is deodorized, it is useful as a nutritional component of feed. Therefore, as one aspect of the present invention, soy sauce can be added to the solid medium from the beginning to culture the microorganism, or soy sauce can be added to the culture after the start of the culture and before the end of the culture. A fermented feed additive containing soy sauce cake containing soy sauce oil deodorized by this method and enriched in fat can be obtained. Furthermore, you may add not only soy sauce oil but fats and oils. Examples of the fats and oils include edible oil, rice oil, coconut oil, fish oil, and Aman oil.

Production of feed Since the feed material of the present invention usually contains about 3% of salt derived from soy sauce cake, the amount added to the feed is limited to less than about 10%. Therefore, the method for producing a mixed feed containing soy sauce cake according to the present invention is characterized by adding the fermented feed containing soy sauce cake according to the present invention in an amount of less than 10% to the feed.

Hereinafter, the present invention will be specifically described with reference to experimental examples and examples.
Example 1. Fermentation test of soy sauce cake with various koji molds Soy sauce cake: wheat bran: 25 g of a medium mixed with water at a weight ratio of 1: 1: 2 was placed in a 500 ml Mericlon flask and autoclaved (121 ° C, 15 1 platinum ear was inoculated with various koji molds (described in Table 1) previously cultured on a PDA slant agar medium and cultured at 30 ° C. for 3 days. The obtained culture was air-dried at 50 ° C. for 6 hours, and then 27 ml of physiological saline was added to 3 g of the culture, homogenized (12,000 rpm, 2 minutes), and the homogenate was filtered through No. 6 filter paper. Got. The filtrate was measured for phytase activity according to the following method.

  That is, 0.1 mL of the above filtrate was added to 0.9 ml of 100 mM acetate buffer (pH 5.7) containing 5 mM sodium phytate solution that had been kept at 37 ° C., and reacted at 37 ° C. for 20 minutes. As a control, the same operation was performed using a 100 mM acetate buffer (pH 5.7) not containing sodium phytate. After completion of the reaction, the liberated phosphoric acid was quantified according to the Fiske Subarrow method. One unit of phytase activity was defined as the amount of enzyme that liberates 1 micromole of inorganic phosphate per minute under the above enzyme activity measurement conditions.

The evaluation method of the soy sauce odor was performed by five panelists, and the odor level when the odor before treatment was set to 5 was evaluated in 6 levels from 5 to 0 according to the strength.
As a result, as shown in Table 1, it was found that phytase was produced in the medium by fermentation with fungi of Neurospora and Rhizopus. Production of xylanase, CMCase, β-lactamase and lipase added as feed additives was also observed. Although all of the deodorizing effects had a slight musty odor, the soy sauce odor did not remain, and after standing at room temperature for 1 month, the return scent was evaluated, but there was no return scent.

Example 2. Acquisition of high phytase activity mutants by mutation treatment of Neurospora sitophila NBRC31635 and Rhizopus oligospora NBRC 8631 A spore suspension of Neurospora sitophila NBRC31635 and Rhizopus oligospora NBRC 8631 was prepared and irradiated with UV according to a conventional method. Performed and induced mutations. The UV-treated spore suspension was smeared on a PDA plate medium, and the grown candidate strain was picked on a PDA slant medium and cultured at 30 ° C. for 3 days. Inoculate one platinum loop of 1 mL of liquid culture medium (0.3% polypeptone, 0.3% yeast extract, 0.3% malt extract, 1% glucose, pH 5.5) in a microbial culture plate and culture at 30 ° C for 1 week. . Using 0.01 mL of the culture supernatant from which the cells had been removed, 0.1 mL of 100 mM acetate buffer (pH 5.7) containing 5 mM sodium phytate solution was added and reacted at 37 ° C. for 20 minutes. After completion of the reaction, free phosphate was analyzed according to the Fiske-Subarrow method, and a high phytase activity mutant was selected visually.

Example 3. Fermentation treatment with soy sauce cake and wheat bran using a mutant strain Soy sauce cake and wheat bran were mixed 1: 3, and an equal amount of water was added (1), soy sauce cake and wheat bran. 10g of each medium (2) mixed in 1: 1 and added with an equal amount of water, put into a 100ml Erlenmeyer flask and sterilized by autoclave, and inoculated with 1 platinum loop of the high phytase activity mutant prepared in Example 2 above And cultured at 30 ° C. for 3 days. After the obtained fermented product was dried by ventilation, the results of measuring the phytase activity (unit / g) according to the method described in Example 1 are shown in the following table. In addition, none of the soy sauce odor was felt at all. Among the obtained high phytase activity mutants, Neurospora sitophila NA38 strain (NITE AP-229), NA37 strain (NITE AP-230), Rhizopus oligospora KT28 strain (NITE AP-232), KT27 strain (NITE AP- 231) was deposited.

Example 4. Fermentation treatment with soy sauce cake and tofu cake using Neurospora sitophila NA38 strain Soy sauce cake and tofu cake were mixed in the proportions shown in the table below, and an equal amount of water was added to the medium (3) to (7). 10 g of each was placed in a 100 ml Erlenmeyer flask, autoclaved, and one platinum loop of Neurospora sitophila NA38 strain was inoculated and cultured at 30 ° C. for 3 days. After carrying out ventilation drying of the obtained fermented material, as a result of evaluating the phytase activity (unit / g) and the soy sauce odor according to the method described in Example 1, as shown in Table 3, according to the blending ratio of the soy sauce cake Although the phytase activity was reduced, it was found that the Neurospora sitophila NA38 strain grew even in the medium of 90% soy sauce cake and exhibited phytase activity and deodorizing effect.

Example 5 Fermentation treatment of soy sauce cake, tofu cake and rice bran added with soy sauce medium Soy sauce cake, tofu cake and rice bran were mixed in the proportions shown in the following table, and medium (8) to (10) added with an equal amount of water. 50 g of each was placed in a 500 ml Mericlon flask and autoclaved. One platinum loop of Neurospora sitophila NBRC31635 was inoculated and cultured at 30 ° C. for 3 days. As for the media (11) and (12), after culturing in the media (8), soy sauce was added 1 hour before the end of the culture. After completion of the culture, ventilation drying was performed at 50 ° C. for 6 hours, and the obtained fermented product was evaluated for phytase activity (unit / g) and soy sauce odor according to the method described in 1 above.

  As a result, as shown in Table 4, even when soy sauce was mixed with the medium, Neurospora sitophila NBRC31635 grew and the odor of soy sauce cake and soy sauce was completely removed. However, phytase activity was significantly reduced. Soy sauce oil was added at the end of enzyme production, left for 1 hour, and dried to remove the odor of soy sauce cake and soy sauce oil without reducing phytase activity.

Example 6 Fermentation treatment with soy sauce cake and tofu cake using Rhizopus oligosporus KT28 Soy sauce cake, tofu cake, rapeseed meal and wheat bran were mixed at a ratio of 5: 10: 3: 2 and equal amounts of water Medium (13), soy sauce cake, tofu cake, and wheat bran in a ratio of 1: 1: 1, and 10g of each medium (14) with an equal amount of water added, placed in a 100ml Erlenmeyer flask and autoclaved Then, one platinum loop of Rhizopus oligosporus strain KT28 was inoculated and cultured at 30 ° C. for 3 days. The obtained fermented product was evaluated for phytase activity and soy sauce odor according to the method described in 1 above, and as a result, 6.2 units / g of the medium (13) and 4.1 units / g of the medium (14) were obtained. . None of them felt a soy odor.

Example 7 : Large-scale fermentation treatment using Neurospora sitophila NBRC31635 (25% soy sauce cake)
Sake made by adding equal amount of water to 100 kg of medium mixed with 25% soy sauce koji, 25% tofu koji, 25% rice koji, 15% rapeseed koji, and 10% corn gluten meal, and sterilized at 100 ° C for 30 minutes After cooling to 30 ° C., Neurospora sitophila NBRC31635 spores were inoculated as seed pods, mixed, and cultured at 30 ° C. for 3 days. During the koji making, care (mixing / stirring) was performed on the first and second days to make it uniform. After completion of the fermentation, the fermented product was air-dried (50 ° C., 6 hours) and pulverized with a pulverizer. The obtained fermented product had a phytase activity of 9.8 units / g. I didn't feel any soy odor. Hereinafter, the fermented feed containing soy sauce cake produced by this method is abbreviated as “S25”.

Example 8. Large-scale fermentation treatment using Neurospora cytophila NBRC31635 (50% soy sauce cake)
Add equal volume of water to 100 kg of medium mixed with soy sauce koji 50%, tofu koji 25%, rice koji 25%, and sterilize at 121 ° C for 15 min. Then, Neurospora sitophila NBRC31635 spores were added as seed pods, mixed, and cultured at 30 ° C. for 3 days. During the koji making, care (mixing and stirring) was performed on the 1st and 2nd days to make it uniform. After completion of the fermentation treatment, the fermented product was dried by ventilation (50 ° C., 6 hours) and then pulverized by a pulverizer. The phytase activity of the obtained fermented product was 3.0 units / g. I didn't feel any soy odor. Hereinafter, the fermented feed containing soy sauce cake produced by this method is abbreviated as “S50”.

Example 9 Large-scale fermentation treatment using Neurospora sitophila NBRC31635 (50% soy sauce + 25% soy sauce)
Add equal volume of water to 100 kg of medium mixed with soy sauce koji 50%, tofu koji 25%, rice koji 25%, and sterilize at 121 ° C for 15 min. Then, Neurospora sitophila NBRC31635 spores were added as seed pods, mixed, and cultured at 30 ° C. for 3 days. During the koji making, care (mixing / stirring) was performed on the first and second days to make it uniform. After confirming the phytase activity, 25 kg of soy sauce oil was added. After 1 hour, the fermented product was air-dried (50 ° C., 6 hours) and pulverized with a pulverizer. The phytase activity of the obtained fermented product was 3.0 units / g. There was no soy sauce odor or soy sauce odor. Hereinafter, the fermented feed containing soy sauce cake produced by this method is abbreviated as S50F25.

Example 10. Effect of soy sauce cake mixed feed using Neurospora sitophila (S25) on pigs Using S25 as described in 6 above, instead of the enzyme agent currently used, phytase produced by Neurospora sitophila is It was verified by the following method whether it could actually be applied as a pig feed. In other words, a test feed containing 5% S25 (phosphorus content 0.45%, phytase activity approximately 500 units) was prepared against a control diet (phosphorus content 0.55%) that did not contain phytase, and the phosphorus digestibility and balance were investigated. did. As a result, the phosphorus digestibility of piglets was significantly higher than that of the control diet (control group 57.6% vs test group 73.1%), and the phosphorus accumulation rate was comparable (control group 5.41g / day vs test). Ward 5.19g / day). This proved the phytase effect of S25 on pigs.

  Next, S25 was added to the general feed to investigate the effect on pig growth and meat quality. Divide 6 pigs with an average weight of about 68kg into 2 wards and test feed (14% crude protein, 77% total digestible nutrients) containing 5% S25 and general feed (crude protein 14%, total digestible nutrient 77%) ) And fed to an average weight of about 112 kg. Feed intake (general feed 2.863kg / day vs S25 test feed 2.966kg / day), daily gain (general feed 0.879kg / day vs S25 test feed 0.968kg / day), feed demand rate (general feed 3.257) vs S25 test feed 3.064) showed better values in the S25 combination test feed group. Therefore, it was suggested that S25 not only has a negative impact on pig growth, but rather may improve.

  In addition, when the meat quality of these test pigs was investigated, all parameters of meat color, back fat melting point, and back fat fatty acid composition were the same as those in the general feed section, and no adverse effects of S25 formulation were observed. In addition, a three-point discrimination sensory test for loin meat and rose meat was performed using the meat of this test pig, but there was no statistical difference. Therefore, it was found that S25 has no adverse effect on pork quality.

Example 11 Effect of Soy Sauce Mixture Feed (S25) Using Neurospora / Sitophila on Chickens Phytase is approved as a feed additive and is widely used in commercial egg-laying hen feeds. Therefore, if S25 is added to the feed, it can be expected as an alternative to phytase as a feed additive. This study investigated whether S25 could be used as a commercial feed additive phytase alternative. The test group is a commercial phytase group adjusted to be 300 units / kg feed for phytase recommended for egg laying hens, and S25 group adjusted to become phytase 294 units / kg feed (containing 3% of S25) And the nutrient components of each test feed were adjusted to be equal.

  The test chickens were single cages in the test windowless poultry house, 12 birds in total, 6 birds in each ward, the test start age was 705, and the test period was 8 weeks. S25 feeding did not adversely affect body weight, feed intake, egg production rate and egg weight. Similarly, the How Unit and egg specific gravity related to egg quality were not adversely affected. Based on the above, if feed laying hens containing 3% S25 and adjusted to phytase 294 units / kg feed, feed adjusted to phytase 300 units / kg feed using commercial phytase It was found that the same result as that obtained when feeding to the laying hen was obtained.

Example 12. Effect of soy sauce cake mixed feed (S50, S50F25) on pigs using Neurospora sitofira
In order to verify the effects of S50 and S50F25, the digestibility of S50 and S50F25 was investigated. A basic feed of 15.8% crude protein and a total digestible nutrient amount of 77.3% and a feed containing 20% each of S50 and S50F25 were fed into this basic feed, and the digestibility of the basic feed, S50 mixed feed, and S50F25 mixed feed was measured. As a result, the TDN of S50 was 63.3% and the TDN of S50F25 was 85.6%. Among them, the digestibility of crude fat was significantly improved (basic feed 58.5% vs. S50 mixed feed 63.7%), and the crude fat digestibility of S50 itself Showed 65-83.7%. A similar trend was also observed in S25 (basic feed 58.7% vs S25 mixed feed 72.7%). Therefore, it was confirmed that what mixed soy sauce cake and other feed raw materials with the method of this invention has the effect of improving the digestibility of crude fat in feed.

Example 13. Effects of soy sauce cake mixed feed (S50, S50F25) on chickens using Neurospora sitophila
In order to be used for chicken feed in both S50 and S50F25, their metabolic energy values (ME, kcal / kg) are indispensable in feed design. Therefore, ME measurement of S50 and S50F25 was performed. The test chickens were caged (8 to 9 / cage) in the test windowless poultry house, and one cage was used as one unit, and a total of 50 chickens, 16 or 17 birds, were used. The test starting date was 56. The test feed acclimatization period was 5 days, and then 2 days was the ME measurement period. ME was the result of adding nitrogen correction to the difference between the total energy intake of feed and the total excretion energy of manure.

  The ME of S50 and S50F25 gave 2735 and 3860 results, respectively. Since S50F25 is superior to S50 in terms of ME as a feed ingredient for chickens, the validity of ME measurement results was investigated by feeding S50F25 to laying hens. The feed had the same nutrient composition design in both the control group and the test group, and 3% S50F25 was used in the test group. The contents of digestible amino acids, effective phosphorus, vitamins, trace minerals, etc. that were not found in the nutritional components of S50F25 were treated as component 0.

  The test start and end dates were 125 and 300, respectively, and the test chickens were caged in a test windowless poultry house (6 birds / cage), and 3 consecutive cages were handled as 1 unit. 144 birds were used. Throughout the test period, S50F25 did not adversely affect feed intake, egg production rate, total production, and feed demand rate, indicating the validity of the ME measurement results of S50F25. From the above, it was confirmed that S50F25 can be used without affecting the egg-laying performance when the ME is treated as 3860 and the use in the feed for laying hens is 3%.

Claims (12)

Solid comprising 90% by weight or less of soy sauce cake and 10% by weight or more of solid feed raw material with respect to the total weight of solid and 80 to 150% by weight of water with respect to the weight of the mixture It belongs to Neurospora sitophila NA38 strain (NITE AP-229), Neurospora sitophila NA37 strain (NITE AP-230) or Rhizopus oligosporus species having phytase-producing ability in the medium. A method for producing a fermented feed containing soy sauce cake, comprising culturing microorganisms and accumulating 2 units / g or more of phytase.   The method according to claim 1, wherein the amount of the soy sauce cake relative to the total weight of the solid is 70% by weight or less, and the amount of the solid feed material relative to the total weight of the solid is 30% or more.   The method according to claim 1, wherein the amount of the soy sauce cake relative to the total weight of the solid is 60% by weight or less, and the amount of the solid feed material relative to the total weight of the solid is 40% or more.   The method according to claim 1, wherein the amount of soy sauce cake relative to the total weight of the solid is 50% by weight or less, and the amount of the solid feed material relative to the total weight of the solid is 50% or more.   The method according to any one of claims 1 to 4, wherein the accumulated amount of phytase is 4 units / g or more.   The method according to any one of claims 1 to 5, wherein the accumulated amount of phytase is 9 units / g or more.   The method according to any one of claims 1 to 6, wherein the accumulated amount of phytase is 15 units / g or more.   The method according to any one of claims 1 to 7, wherein the Rhizopus oligosporus is Rhizopus oligosporus KT28 strain (NITE AP-232) or Rhizopus oligosporus KT27 strain (NITE AP-231). . The solid feed material is wheat bran, rice bran, tofu koji, oil koji, soybean koji, koji koji, corn gluten meal, raw ore produced as a by-product during soy sauce production, or grilled ori produced as a by-product during soy sauce production, or two of these The method according to any one of claims 1 to 8 , which is a mixture. Furthermore, the addition of fats and oils, the method according to any one of claims 1-9. 11. The method according to claim 10, wherein the fats and oils are soy sauce oil. A fermented feed containing soy sauce cake is produced by the method according to any one of claims 1 to 11 , and the fermented sample is added to the feed in an amount of 10% by weight or less based on the total weight of the feed. The manufacturing method of mixed feed containing soy sauce cake.