阅读说明：本技术 发酵大豆粕及其制备方法 (Fermented soybean meal and preparation method thereof ) 是由 李钟和 李赫基 于 2019-07-17 设计创作，主要内容包括：本发明涉及一种发酵大豆粕及其制备方法,更具体地,涉及一种使用兼性厌氧大豆粕发酵微生物制备发酵大豆粕的方法、由其制备的发酵大豆粕和含有该发酵大豆粕的饲料组合物。所述发酵大豆粕的制备方法包括：通过用提取溶剂提取生大豆粕来获得大豆粕提取物溶液和残余大豆粕的步骤,和通过使用发酵大豆粕的微生物对包含选自由生大豆粕和残余大豆粕组成的组中的一种或多种的发酵原料进行固体培养的步骤。此外,本发明涉及一种新的肠球菌属菌株,更具体地,涉及屎肠球菌菌株。(The present invention relates to fermented soybean meal and a preparation method thereof, and more particularly, to a method for preparing fermented soybean meal using facultative anaerobic soybean meal fermenting microorganisms, fermented soybean meal prepared thereby, and a feed composition containing the fermented soybean meal. The preparation method of the fermented soybean meal comprises the following steps: a step of obtaining a soybean meal extract solution and residual soybean meal by extracting raw soybean meal with an extraction solvent, and a step of subjecting a fermentation raw material containing one or more selected from the group consisting of raw soybean meal and residual soybean meal to solid culture by using a microorganism that ferments soybean meal. Furthermore, the present invention relates to a novel enterococcus strain, more specifically, to an enterococcus faecium strain.)

1. A method for preparing fermented soybean meal, comprising:

a step of obtaining a soybean meal extract solution and residual soybean meal by extracting raw soybean meal with an extraction solvent, and

a step of subjecting a fermentation raw material containing one or more selected from the group consisting of raw soybean meal and residual soybean meal to solid culture by using a microorganism that ferments soybean meal.

2. The production method according to claim 1, wherein the microorganism fermenting soybean meal is facultative anaerobic lactic acid bacteria, and the step of solid culture does not include an aerobic process.

3. The production method according to claim 1, wherein the microorganism fermenting soybean meal is added as a seed culture solution obtained by inoculating the microorganism into a fermentation raw material in an amount of 0.000001 to 10 wt% based on 100 wt% of a soybean meal extract solution and cultured in a liquid at a temperature of 20 to 50 ℃.

4. The preparation method according to claim 1, wherein the extraction solvent is one or more selected from the group consisting of water and alcohols of 1 to 6 carbon atoms, and is used in a weight ratio of 1 to 10 times that of the raw soybean meal.

5. The production method according to claim 1, wherein the temperature of the extraction solvent is 20 to 70 ℃ and the pH is 2 to 8.

6. The production method according to claim 1, wherein the residual soybean meal having a moisture content of 80 w/w% or less is obtained by extracting the raw soybean meal with an extraction solvent and then separating by a centrifugal process.

7. The production method according to claim 1, wherein the fermentation raw material is a mixture of residual soybean meal and raw soybean meal, and the crude protein content or the anti-nutritional factor content of the fermented soybean meal is controlled by adjusting a mixing ratio.

8. The preparation method of claim 7, wherein the anti-nutritional factor is one or more selected from the group consisting of trypsin inhibitor, β -conglycinin, indigestible oligosaccharide, hemagglutinin (lectin), saponin and tannic acid.

9. The production method according to claim 1, wherein the fermentation raw material is a mixture in which the raw soybean meal and the residual soybean meal are mixed in a weight ratio of 1: 10 to 10: 1.

10. The preparation method according to claim 1, wherein the fermentation feedstock is obtained from soybean meal and comprises a crude protein content of 20 to 48% (w/w) and an indigestible oligosaccharide content of 0.6 to 1.7 (w/w%).

11. The method of manufacturing of claim 1, comprising:

a step of obtaining a soybean meal extract solution and residual soybean meal by extracting raw soybean meal with an extraction solvent;

a step of preparing a fermentation raw material in which the crude protein content is 20 to 48% (w/w) and the indigestible oligosaccharide content is 0.6 to 1.7 (w/w%) by mixing the raw soybean meal and the residual soybean meal, and

a step of subjecting the fermentation raw material to solid culture by using a microorganism that ferments soybean meal.

12. The production method according to claim 1, wherein the soybean meal fermenting microorganism is one or more selected from the group consisting of Enterococcus (Enterococcus sp.), Weissella (Weissella sp.), and Lactobacillus (Lactobacillus sp.).

13. The production method according to claim 7, wherein the microorganism fermenting soybean meal is one or more selected from the group consisting of: enterococcus faecium (Enterococcus faecium), Enterococcus faecalis (Enterococcus faecium), Weissella gaurea (Weissella koreensis), Pediococcus pentosaceus (Pediococcus pentosaceus), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus lactis (Lactobacillus lactis), Lactobacillus reuteri (Lactobacillus reuteri), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus fermentum (Lactobacillus ferum), Lactobacillus bulgaricus (Lactobacillus bulgaricus) and Lactobacillus acidophilus (Lactobacillus acidophilus).

14. The preparation method according to claim 1, wherein the fermented soybean meal has a crude protein content of more than 46% (w/w) to less than 80% (w/w).

15. A soybean meal fermentation product obtained by fermenting a fermentation raw material comprising one or more selected from the group consisting of raw soybean meal and residual soybean meal using a facultative anaerobic soybean meal fermenting microorganism,

wherein the trypsin inhibitor is present in an amount of 0.0001 to 8(mg/g), the beta-conglycinin is present in an amount of 0 to 70000(ppm), or the indigestible oligosaccharide is present in an amount of 0.0001 to 1.7 (w/w%), and the crude protein content is higher than 46% to 80% (w/w).

16. The soybean meal fermentation product of claim 15, wherein the fermentation feedstock comprises one or more selected from the group consisting of raw soybean meal and residual soybean meal, the residual soybean meal being a solid component obtained by removing a soybean meal extract solution in a solvent extract of soybean meal.

17. The soybean meal fermentation product of claim 15, wherein the fermentation feedstock is obtained from soybean meal and has a crude protein content of 20 to 48% (w/w) and an indigestible oligosaccharide content of 0.6 to 1.7 (w/w%).

18. An animal feed composition comprising the soybean meal fermentation product of claim 15.

19. The feed composition of claim 18, wherein the animal is one or more selected from the group consisting of swine, cattle, chicken, duck, goat, sheep, dog, and cat.

20. The feed composition of claim 18, wherein the soybean meal fermentation product has a crude protein content of 48 to 53% (w/w) and the animal is an adult animal.

21. The feed composition of claim 18, wherein the soybean meal fermentation product has a crude protein content of 50 to 60% (w/w) and the animal is a piglet or a chicken.

22. The feed composition of claim 18, wherein the soybean meal fermentation product has a crude protein content of 53 to 65% (w/w) and the animal is a fish.

23. The production method according to claim 1, wherein, in the fermented soybean meal, the content of the protein having a molecular weight of less than 25kD is 25 to 99.9 wt%, the content of the protein having a molecular weight of 25 to less than 50kD is 0.01 to 60 wt%, and the content of the protein having a molecular weight of 50kD or more is 0.01 to 30 wt%, based on 100 wt% of the total protein content in the fermented soybean meal.

24. An Enterococcus faecium (Enterococcus faecium) strain having an optimum growth temperature of 40 to 45 ℃ and a crude protein content of more than 49 to 80% (w/w) in a soybean meal fermentation product obtained by fermentation with the strain.

25. The strain of claim 24, having one or more characteristics selected from the group consisting of (1) to (6) below:

(1) the soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain has a trypsin inhibitor content of less than 3.5mg/g,

(2) the beta-conglycinin of the soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain is 10000ppm or less,

(3) the soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain has an indigestible oligosaccharide content of 0.1 w/w% or less,

(4) the lactic acid concentration of the soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain was 3 w/w% or more,

(5) the pepsin digestibility of the soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain is 90 w/w% or more, an

(6) In the protein distribution of a soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain, the content of protein having a molecular weight of less than 25kD is 30 to 99.9 wt%, the content of protein having a molecular weight of 25 to less than 50kD is 0.01 to 60 wt%, and the content of protein having a molecular weight of 50kD or higher is 0.01 to 25 wt%, based on 100 wt% of the total protein content in the fermented soybean meal.

26. The strain of claim 24, wherein the strain is enterococcus faecium SLB130 strain with deposit number KCTC13566 BP.

27. The strain of claim 24, wherein the soybean meal fermentation product is obtained by fermenting one or more selected from the group consisting of raw soybean meal and residual soybean meal obtained by extracting the raw soybean meal with an extraction solvent.

28. A composition for fermenting soybean meal, comprising one or more selected from the group consisting of microbial cells of enterococcus faecium SLB130 strain, a culture product of the strain, a lysate of the strain, and an extract of the strain.

29. The composition of claim 28, wherein the strain is cultured in a soybean meal extract solution obtained by extracting soybean meal with an extraction solvent.

30. The composition of claim 28, wherein the composition for fermenting soybean meal ferments one or more selected from the group consisting of raw soybean meal and residual soybean meal obtained by extracting the raw soybean meal with an extraction solvent.

31. A method for producing fermented soybean meal, comprising the step of preparing a fermentation raw material selected from one or more of the group consisting of raw soybean meal and residual soybean meal obtained by extracting the raw soybean meal with an extraction solvent; and

a step of solid culture by inoculating the enterococcus faecium SLB130 strain to a fermentation raw material.

32. The production method according to claim 31, further comprising a seed culture step of culturing a strain in a soybean meal extract solution obtained by extracting the raw soybean meal with an extraction solvent before the fermentation step is performed.

33. The method of claim 31, wherein the fermentation feedstock comprises 20 to 48% (w/w) crude protein and 0.6 to 1.7 (w/w) indigestible oligosaccharide.

34. The method of claim 31, wherein the fermented soybean meal has a crude protein content of greater than 46% (w/w) to less than 80% (w/w).

35. The preparation method of claim 31, wherein the fermentation raw material is a mixture in which raw soybean meal and residual soybean meal are mixed in a weight ratio of 1: 10 to 10: 1.

Technical Field

The present invention relates to fermented soybean meal, a method of preparing fermented soybean meal, and a feed composition comprising fermented soybean meal. In addition, the present invention relates to a novel Enterococcus sp strain, a method of preparing fermented soybean meal using the same, fermented soybean meal prepared by the method, and a feed composition comprising the fermented soybean meal.

Background

The defatted soybean meal accounts for 60 percent of the raw materials of the world plant protein feed on the production basis. Since the negative prospects of animal protein sources due to outbreaks of bovine spongiform encephalopathy and the like have recently increased, the demand for defatted soybean meal is expected to continue to increase in the future.

Although activators, antibiotics, antibacterial agents, growth hormone drugs, and the like can be used to improve the productivity of livestock, the use of microbial agents such as probiotics has recently been increasing due to the problems of increased resistance of pathogenic bacteria caused by antibiotics and the like and the retention in livestock products and the like. The microbial preparation can be used for promoting enzyme secretion in animals, improving digestibility of indigestible fibrin, and reducing anti-nutritional factors.

Among soybean components, anti-nutritional factors such as trypsin inhibitor are unstable under heating and thus are mostly inactivated during the production of soybean meal, but for indigestible oligosaccharides such as raffinose and stachyose which are stable under heating, a method of microbial fermentation or enzymatic decomposition is generally used.

A solid substrate fermentation method is generally used worldwide for fermenting soybean meal, and an aerobic fermentation method using an aerobic microorganism, which generally uses a fungus represented by Aspergillus sp and a bacterium represented by Bacillus sp as a microorganism and forcibly supplies oxygen, is mainstream. This aerobic solid substrate fermentation method has a problem that oxygen supply is inevitably required, and therefore the possibility of contamination is high, and a large equipment investment such as a koji-making machine or the like is required, and the scale-up of a plant is not easy. Therefore, solid substrate fermentation processes using anaerobic bacteria are considered as an alternative that can overcome these problems of aerobic processes.

Anaerobic bacteria lactic acid bacteria convert indigestible oligosaccharides, which are one of representative anti-nutritional factors, into beneficial substances such as lactic acid and the like, and can be used for livestock due to active metabolism of amino acids, and also produce various bioactive substances. In particular, since lactic acid secreted during fermentation of lactic acid bacteria can inhibit proliferation of other microorganisms, there is an advantage in that contamination is minimized. However, the conventional anaerobic soybean meal fermentation process generally uses lactic acid bacteria of the genus Lactobacillus (Lactobacillus sp), which has disadvantages of taking a long time and requiring excessive equipment to ensure anaerobic conditions because the lactic acid bacteria are slower than aerobic bacteria in growth and development and fermentation processes.

Disclosure of Invention

Technical problem

The present invention provides a method for preparing fermented soybean meal using soybean meal comprising sugars and some proteins in the soybean meal and a microorganism fermenting the soybean meal.

It is another object of the present invention to provide a method for controlling the crude protein content in fermented soybean meal by preparing a soybean meal extract solution and residual soybean meal using an extraction solvent and using them as fermentation raw materials.

Further, the present invention provides fermented soybean meal having an appropriate crude protein content according to the target livestock species and the growing part of livestock, and a feed composition comprising the fermented soybean meal.

In addition, the present invention prepares a soybean meal extract solution containing sugars and some proteins in soybean meal, and selects a new strain showing a high specific growth rate when used as a medium, thereby creating an extraction and fermentation process of soybean meal based on the new strain.

Technical scheme

The soybean meal itself is used as feed or the like, but may be prepared as soybean meal by treating raw soybean meal with an enzyme or a microorganism, or as residual soybean meal from which unnecessary components are removed with an extraction solvent, for the purpose of promoting enzyme secretion in an animal body, improving the digestibility of indigestible fibrin by a fermentation process, reducing anti-nutritional factors, or the like. The fermentation raw material is prepared using an extraction solvent in order to remove anti-nutritional factors of soybean meal, and to remove proteins by selectively extracting components other than proteins to concentrate the crude protein content. The anti-nutritional factors include trypsin inhibitor, indigestible oligosaccharide such as raffinose and stachyose, etc

One embodiment of the present invention relates to a method of preparing fermented soybean meal by preparing a soybean extract solution and residual soybean meal using an extraction solvent, and by using them as fermentation raw materials and using microbial fermentation of the fermented soybean meal. The soybean meal fermenting microorganism may be Enterococcus faecium (Enterococcus faecium) SLB 130.

In addition, the residual soybean meal obtained by extracting raw soybean meal with an extraction solvent has a high crude protein content, and can be used by adjusting the crude protein content to be suitable for the purpose of use. Thus, the crude protein content of the soybean meal fermentation product can be controlled by reducing the crude protein content by using the residual soybean meal obtained after extraction or mixing raw soybean meal into the residual soybean meal. For example, solid culture using only residual soybean meal as a fermentation feedstock is suitable for producing a crude protein product at a relatively high concentration, and it can be used as a substitute for fish meal used in fish feed. The feed composition may be used as a feed composition for adult livestock, piglets or hatched fish depending on the crude protein content.

In addition, depending on the respective differences in the desired anti-nutritional factor level and crude protein content of livestock, it is important to achieve appropriate anti-nutritional factor levels and crude protein contents to produce fermented soybean meal of the quality required for each target livestock.

Therefore, another example of the present invention relates to a method of controlling a mixing ratio of residual soybean meal and raw soybean meal, which produces fermented soybean meal in a quality required for each target livestock, taking into consideration the levels of anti-nutritional factors and crude proteins required for the livestock to be used.

For example, for fermented soybean meal suitable for hatching fish such as shrimp, abalone, salmon, or the like, fermented soybean meal may be prepared by using residual soybean meal as a fermentation raw material. In other words, the residual soybean meal after solid culture extraction is suitable for producing a high-concentration crude protein product, and it can be used as a substitute for fish meal for hatching fish. Specifically, the fermented soybean meal preferably has an anti-nutritional factor content of less than 0.03% in the case of indigestible oligosaccharides, as low as 200ppm in the case of β -conglycinin, and as high as 58% or higher in the case of crude protein content. Specifically, in fermented soybean meal suitable for fish or hatched fish, the crude protein content may be 53 to 65% (w/w), above 53 to 65% (w/w), 54 to 65% (w/w), 55 to 65% (w/w), 56 to 65% (w/w), above 56 to 65% (w/w), 57 to 65% (w/w), 58 to 65% (w/w), 59 to 65% (w/w), or 60 to 65% (w/w).

For example, since the digestive organs of adult livestock such as sows, growing pigs, or domestic cattle are well developed, it does not matter that the concentration of antinutritional factors in fermented soybean meal is relatively high, it does not matter that the absorption rate of amino acids is higher than that of general soybean meal and the crude protein content is 50% or more. Thus, fermented soybean meal for adult livestock can be prepared by using raw soybean meal. Specifically, in fermented soybean meal suitable for adult livestock such as sows, growing pigs, or domestic cattle, etc., the crude protein content may be 48 to 53% (w/w), more than 48 to less than 51% (w/w), more than 48 to 50% (w/w), 49 to 53% (w/w), 49 to 51% (w/w), 49 to less than 51% (w/w), 49 to 50% (w/w), more than 49% to less than 53% (w/w), more than 49% to 51% (w/w), or more than 49% to less than 51% (w/w).

For example, in the case of young livestock such as piglets, chickens and the like, the effect of the anti-nutritional factor is smaller than that of hatched fish, but it is affected more by the anti-nutritional factor than adult livestock. Thus, in fermented soybean meal for young animals, the content of anti-nutritional factors is less than 0.05% in the case of non-digestible oligosaccharides and less than 500ppm in the case of beta-conglycinin. Thus, fermented soybean meal for young livestock can be prepared by mixing residual soybean meal and raw soybean meal, and the mixing ratio thereof may be 1: 1 to 1: 1.4. Specifically, in fermented soybean meal for young animals such as piglets, hatchlings and the like, the crude protein content may be more than 50 to less than 60% (w/w), more than 50 to 59% (w/w), more than 50 to 58% (w/w), more than 50 to 57% (w/w), more than 50 to 56% (w/w), more than 50 to less than 56% (w/w), more than 50 to 55% (w/w), more than 50 to 54% (w/w), more than 50 to 53% (w/w), 51 to less than 60% (w/w), 51 to 59% (w/w), 51 to 58% (w/w), 51 to 57% (w/w), 51 to 56% (w/w), 51 to 55% (w/w), 51 to less than 55% (w/w), 51 to 55% (w/w), 51 to 54% (w/w), 51 to 53% (w/w), more than 51 to less than 60% (w/w), more than 51 to 59% (w/w), more than 51 to 58% (w/w), more than 51 to 57% (w/w), more than 51 to 56% (w/w), more than 51 to less than 56% (w/w), more than 51 to 55% (w/w), more than 51 to 54% (w/w), more than 51 to 53% (w/w), 52 to less than 60% (w/w), 52 to 59% (w/w), 52 to 58% (w/w), 52 to 57% (w/w), 52 to 56% (w/w), 52 to less than 56% (w/w), 52 to 55% (w/w), 52 to 54% (w/w), 52 to 53% (w/w), 53 to less than 60% (w/w), 53 to 59% (w/w), 53 to 58% (w/w), 53 to 57% (w/w), 53 to 56% (w/w), 53 to less than 56% (w/w), 53 to 55% (w/w), or 53 to 54% (w/w).

Accordingly, one embodiment of the present invention provides: a method for producing a fermented product of soybean meal by fermenting one or more fermentation raw materials selected from the group consisting of raw soybean meal and residual soybean meal with a microorganism that ferments soybean meal; or a method for controlling the crude protein content of a soybean meal fermentation product. The soybean meal fermenting microorganism can be enterococcus faecium SLB130 strain.

The present invention provides a method for producing fermented soybean meal with high cost competitiveness in the animal feed market by an anaerobic fermentation process technique that can produce fermented soybean meal with relatively simple process equipment and low cost by using facultative anaerobic lactic acid bacteria as soybean meal fermenting microorganisms, as an alternative to conventional aerobic fermentation processes that require a large amount of equipment and operating costs. Since the fermentation time is rather short and the possibility of contamination is low, the fermentation process in the present invention can be stably operated. The facultative anaerobic lactic acid bacteria may be enterococcus faecium SLB130 strain.

Since the present invention prepares a soybean meal extract solution and residual soybean meal using an extraction solvent and prepares a soybean meal fermentation product by performing fermentation using them as a fermentation raw material, in the process of the present invention, indigestible oligosaccharides that significantly reduce the feed efficiency of soybean meal by interfering with digestive absorption of livestock are excellently removed. In addition, fermented soybean meal distinguished according to livestock species and targets can be produced by using the extraction process.

The present invention uses the waste soybean meal extract solution as a lactic acid bacteria culture medium, so that the cost can be reduced, and an environment-friendly method without process waste is also provided.

Further, the present invention proposes a technical basis which can obtain products such as lactic acid not only by using a soybean meal extract solution as a lactic acid bacteria culture medium but also by culturing microorganisms, and which can produce various fermentation products having industrial value by fermenting the soybean meal extract solution. Lactic acid required for preparing PLA is mainly produced by fermentation using lactic acid bacteria, and when the extract solution of the present invention and lactic acid bacteria are used, an economical lactic acid preparation process can be performed.

In the present invention, it was confirmed that the growth and development of the lactobacillus species of Enterococcus (Enterococcus sp.) in the soybean meal extract solution selected in the present invention were significantly superior compared to other lactobacillus species or bacillus subtilis (bacillus subtilis) etc. (example 3). In addition, it is well known that such lactic acid bacteria have been a type of lactic acid bacteria that increase the rate of digestive absorption by being added to feed and marine products of livestock or animals as pets. The enterococcus faecium SLB130 strain has the advantages of active growth and development in soybean meal extract solution and easy preparation.

In the present invention, a necessary and sufficient fermentation effect for preparing fermented soybean meal can be obtained by a simple and economical method of inoculating and mixing microbial fermented soybean meal to a fermentation raw material and fixing. However, for this result, it is necessary to inoculate a sufficiently large amount of the soybean meal fermenting microorganisms, which can be solved by using a high-concentration culture solution prepared from a soybean meal extract solution.

In the present invention, a high concentration culture method using a soybean meal extract solution as a culture medium has been developed to produce a large amount of lactic acid bacteria inoculum. For this reason, strains showing high levels of growth and development in the soybean meal extract solution prepared by the above-described method at a temperature of 40 ℃ or higher were studied. Specifically, the growth and development rates of microorganisms at a temperature of 40 ℃ or more were measured by using a soybean extract solution, to which no substance was added, as a culture medium. This is because, since a large amount of water is added for the activity of the microorganism in the actual solid fermentation of the soybean meal, the environment that the microorganism experiences is close to a liquid state in which the aqueous organic matter of the soybean meal is largely dissolved, and furthermore, this is for obtaining a large amount of lactic acid bacteria inoculum with the soybean meal extract solution. As a medium for culturing the strain, MRS medium was used as a selection medium.

The present invention relates to enterococcus faecium SLB130 strain.

This strain can be deposited under the accession number of KCTC13566 BP.

According to another embodiment of the present invention, the present invention relates to a method for preparing fermented soybean meal, comprising the step of inoculating enterococcus faecium SLB130 strain into soybean meal.

According to another embodiment of the present invention, there is provided a method for producing a soybean meal fermentation product by solid fermentation, characterized by comprising the step of inoculating enterococcus faecium SLB130 strain into soybean meal.

According to another example of the present invention, there is provided a method of preparing a soybean meal fermentation product, comprising: a step of preparing one or more fermentation raw materials selected from the group consisting of raw soybean meal and residual soybean meal obtained by extracting raw soybean meal with an extraction solvent; a seed culture step of inoculating the enterococcus faecium SLB130 strain into a soybean meal extract solution obtained by extracting raw soybean meal with an extraction solvent and performing seed culture; and a step of performing solid culture by inoculating the seeds into the fermentation raw material.

The raw material culture medium for seed culture or solid culture may be (1) raw soybean meal, (2) soybean meal extract solution, (3) residual soybean meal after extraction, or (4) a mixture of raw soybean meal and residual soybean meal after extraction.

Another example of the present invention is to extract certain components from soybean meal and culture lactic acid bacteria by using the extract, and then inoculate the cultured lactic acid bacteria into the soybean meal for solid culture. In addition, it may include a step of recycling the culture solution of lactic acid bacteria.

Hereinafter, the present invention will be described in more detail.

In the present invention, the "soybean meal" may be one or more selected from the group consisting of (1) raw soybean meal, (2) soybean extract solution, and (3) residual soybean meal after extraction.

In the present invention, the term "raw soybean meal" refers to soybean meal prior to extraction of the soybean meal with a solvent. Raw soybean meal contains a large amount of protein, and therefore it is the main protein source of livestock feed. The soybean meal may refer to all common defatted soybean meal and soybean meal in general, but is not limited thereto.

In the present invention, the term "soybean meal extract" refers to a soybean extract obtained by extracting raw soybean meal with a solvent, and the soybean meal extract includes a "soybean extract solution" and "residual soybean meal" remaining after removing the "soybean extract solution".

By performing the solvent extraction process, the anti-nutritional factors and saccharides contained in the raw soybean meal can be reduced or removed. In raw soybean meal, polysaccharides such as stachyose, raffinose, and the like and various carbohydrates are contained, which are mostly aqueous, and include an antinutritional factor (hereinafter referred to as ANF) that interferes with digestion of livestock. In addition, the crude protein content in the raw soybean meal may be concentrated by the extraction process, so that the crude protein content in the residual soybean meal and the soybean meal fermented using the same may be increased.

In the present invention, "anti-nutritional factors" may include all substances that interfere with the digestion of livestock, and may include all substances contained in soybean meal or fermented soybean meal that are negative for digestion. For example, the anti-nutritional factor may be one or more selected from the group consisting of trypsin inhibitor, beta-conglycinin, indigestible oligosaccharide, hemagglutinin (lectin), saponin, and tannic acid.

In order to remove the antinutritional factors of the soybean meal and concentrate the crude protein content, enzyme treatment or fermentation should be performed after removing components other than proteins. For example, when indigestible oligosaccharides such as raffinose and stachyose, which are anti-nutritional factors present in soybean meal, are first extracted and removed, the anti-nutritional factors are removed while the crude protein content of the residual soybean meal remaining after extraction is increased. In other words, the crude protein content in the residual soybean meal and the soybean meal fermented using it can be controlled by the extraction process of the raw soybean meal.

The extraction of soybean meal of the present invention uses a method of efficiently extracting non-protein components in soybean meal by using acidic water as a solvent. This is the following principle: since most of proteins in soybean meal have an isoelectric point (hereinafter, pI) of 4.5, when extraction is performed by using weakly acidic water having a pH of less than 4.5 as a solvent, proteins precipitate and are thus not extracted, and aqueous oligosaccharides and carbohydrates having no influence on the pH are extracted.

In order to ferment soybean meal in large quantities, large quantities of inoculum are required. This is particularly the case in solid fermentation, which makes it difficult to control the fermentation environment. In general, since it is not difficult to construct a uniform liquid fermentation environment, temperature and pH control, oxygen supply (or oxygen blocking), and the like are easy, a large amount of inoculum is not required. Typically, about 10 per ml is used⁶To 10⁷cfu (colony forming units) or inoculum size of 1 to 5 vol%.

In particular, to use lactic acid bacteria for fermenting soybean meal, the use of a sufficiently large amount of inoculum has various advantages. Since the growth and development rate of lactic acid bacteria that grow and develop anaerobically (or facultative anaerobiosis) is slower than that of aerobic bacteria or yeast, fungi, etc., a larger amount of inoculum is required to ferment soybean meal efficiently.

When selecting a fermenting microorganism, a microorganism having an excellent decomposing ability is generally selected by targeting most or the main component of the organic components possessed by the object. Therefore, when studying microorganisms for fermenting soybean meal, a common method is to find microorganisms that produce enzymes that decompose major constituent sugars in soybean meal, particularly raffinose and stachyose, which are indigestible oligosaccharides, and produce a large amount of enzymes that decompose soybean meal proteins. This is because representative antinutritional factors (trypsin inhibitor (TI) and β -conglycinin, etc.) of soybean are composed of proteins, and it is beneficial to decompose proteins at the small size level of proteins or peptides, so that livestock can smoothly digest soybean proteins having a relatively large molecular weight. In addition, a secondary condition is a microorganism that can cause fermentation by adapting to the soybean meal solid fermentation conditions (e.g., moisture content, temperature, pH, etc.). Bacteria in general, which exist in the general natural environment but not in the intestinal tract, have many beneficial points of fermentation in particular because the optimal temperature for growth and development is high, because many bacteria cannot grow at 40 ℃ or higher. Furthermore, since the body temperature is about 41 ℃ in the case of chickens or cattle, it may be a characteristic of lactic acid bacteria that are beneficial for survival and proliferation in the actual intestinal tract of livestock.

In the present invention, fermented soybean meal is prepared by obtaining an extract from soybean meal, selecting and culturing lactic acid bacteria that grow well under anaerobic conditions into the extract, and then anaerobically fermenting the soybean meal with the cultured lactic acid bacteria. Hereinafter, soybean meal is a common name for both ordinary defatted soybean meal and soybean meal.

Soybean meal contains a large amount of protein and is therefore the main protein source of livestock feed. A soybean protein concentrate (hereinafter, referred to as SCP) in which an anti-nutritional factor component is removed from soybean meal and a protein fraction is increased is a high-grade product used for feeds for weaned piglets and the like, and a mixture of ethanol and water is used as an extraction solvent. However, this process has the disadvantage that the alcohol used as solvent requires expensive recovery equipment. In addition, residual alcohol components may inhibit the growth and development of microorganisms. In the present invention, the non-protein components of the soybean meal are extracted by using acidic water as a solvent instead of alcohol having such a disadvantage.

Since the isoelectric point (hereinafter, referred to as pI) of the protein in the soybean meal is about 4.5, when extraction is performed using weakly acidic water having a pH of less than 4.5 as a solvent, the protein precipitates and is thus not extracted, and aqueous oligosaccharides and carbohydrates having no influence on the pH are extracted. In order to ferment soybean meal in large quantities, large quantities of inoculum are required. Since solid fermentation is difficult to create a uniform environment, unlike liquid fermentation, it is necessary to maintain sterilization and fermentation conditions. Therefore, more inoculum should be used than in liquid fermentation, which necessarily increases the process cost.

Generally, during lactic acid bacteria fermentation, lactic acid is secreted, which shows the effect of lowering the pH of the medium and finally acts to prevent the proliferation of other contaminants. On the other hand, since the growth and development rate of lactic acid bacteria that grow and develop anaerobically (or facultative anaerobically) is lower than that of aerobic bacteria or yeast, fungi, etc., a larger amount of inoculum is required to effectively ferment soybean meal by using lactic acid bacteria. Bacteria in general, which exist in the general natural environment but not in the intestinal tract, have many beneficial points of fermentation in particular because the optimal temperature for growth and development is high, because many bacteria cannot grow at 40 ℃ or higher. Furthermore, since the body temperature is about 41 ℃ in the case of chickens or cattle, it may be a characteristic of lactic acid bacteria that are beneficial for survival and proliferation in the actual intestinal tract of livestock.

In the present invention, in order to produce a large amount of lactic acid bacteria inoculum, a high concentration culture method using a soybean meal extract solution as a culture medium was developed. For this reason, first, lactic acid bacteria showing high level growth and development in the soybean meal extract solution prepared by this method at 40 ℃ or more were studied.

In the present invention, the growth and development rate of microorganisms at 40 ℃ or more is measured by using a soybean meal extract solution without any substance added thereto as a liquid medium. This is because, since a large amount of water is added for the activity of the microorganism in the actual solid fermentation of the soybean meal, the environment that the microorganism experiences is close to a liquid state in which the aqueous organic matter of the soybean meal is largely dissolved, and furthermore, this is for obtaining a large amount of lactic acid bacteria inoculum with the soybean meal extract solution. As a medium for culturing lactic acid bacteria, MRS medium was used as a selection medium.

The lactobacillus strain of the present invention effectively decomposes and utilizes soybean meal organic substances such as oligosaccharides, carbohydrates, proteins, etc., particularly sucrose, stachyose, raffinose, etc., which are not well used as carbon sources by common microorganisms. As such, by culturing microorganisms using an extract from which oligosaccharides that reduce the performance of soybean meal as an anti-nutritional factor are removed as a culture medium, not only can the cost of preparing an inoculum for fermentation of soybean meal be reduced, but also new values can be generated by additionally producing industrial lactic acid bacteria powder, or lactic acid, amino acids, or the like.

In the present invention, a continuous culture method has been invented to culture selected lactic acid bacteria in a soybean meal extract solution at a high concentration. The continuous culture method can continuously obtain culture products by adding a new medium to a small-sized culture tank instead of the batch culture method.

On the basis of the very high specific growth rate of the selected lactic acid bacteria, the continuous culture method of the present invention does not use sodium hydroxide or lactic acid neutralizing agent such as alkali like calcium carbonate, but allows the soybean meal extract solution to be automatically added, thereby maintaining a certain pH value. The method can increase the recyclability of resources and can simplify the process in that no additional additives are used.

In order to remove the antinutritional factors of soybean meal and concentrate the crude protein content, after removing components other than proteins by selective extraction, enzyme treatment or fermentation should be performed. This is because most of the soybean meal antinutritional factors have a protein structure. Recently, such objects have been achieved by fermentation, generally using bacteria of the genus Bacillus or fungi of the genus Aspergillus (Aspergillus sp.) or yeasts of the genus Saccharomyces (Saccharomyces cerevisiae/Saccharomyces cerevisiae).

When fermentation is performed using such aerobic microorganisms, initial investment costs, running costs, cost price, and the like increase, and most importantly, enlargement of the production scale is not easy. In other words, in the aerobic process, a great deal of effort is required to provide a uniform and sufficient amount of oxygen, and thus, the thickness to be stacked is limited, and thus, a large amount of space is required in facilities, and it is relatively inefficient to use resources such as water vapor, water, air, heat, and the like.

In contrast, anaerobic processes do not require the supply of oxygen, and therefore it is a great advantage that fermentation conditions can be maintained by stacking without the need for expensive/costly koji-making machines. Since heat is generated during the fermentation process, no additional facilities and energy are required to maintain the fermentation temperature. In addition, when lactic acid bacteria, which are representative anaerobic microorganisms, are used, there is a great advantage in that lactic acid produced during fermentation lowers the pH of fermented soybean meal, so that contamination by various bacteria can be prevented.

However, anaerobic fermentation using lactic acid bacteria generally has the big disadvantage of slow rate. To overcome this disadvantage, in the present invention, lactic acid bacteria (SLB130) having a rapid growth and development rate during fermentation of soybean meal are selected.

Lactic acid bacteria have excellent availability to soybean meal oligosaccharides, however, have a disadvantage of relatively low protease activity, but the lactic acid bacteria of the present invention exhibit excellent protein decomposition ability and thus have sufficient competitiveness. As a result of identifying the lactic acid bacterium of the present invention, it was identified as an Enterococcus faecium (Enterococcus faecium) strain.

In the present invention, necessary and sufficient fermentation effects required for preparing fermented soybean meal can be obtained by a simple and economical method of fixing for 10 to 24 hours after inoculating and mixing lactic acid bacteria into the extracted soybean meal. However, this result requires a sufficiently large amount of inoculum, but can be solved by using a high concentration of lactic acid bacteria culture solution prepared by the above-mentioned extraction solution.

The extract solution produced in the extraction process of the present invention contains a large amount of oligosaccharides and a small amount of proteins, and thus can be used as a culture medium for microorganisms. Since the extract solution contains a large amount of raffinose, stachyose, and the like, a discardable resource can be recovered when the extract solution is used for culture or fermentation of lactic acid bacteria that can sufficiently utilize it.

The lactic acid bacteria of the genus enterococcus selected in the present invention are well-known lactic acid bacteria which are added to feed for livestock or animals as pets, marine products, and the like, improve digestibility, and are well-known as inoculated microorganisms of silage or forage. Therefore, in addition to the inoculated microbial cells required for the soybean meal fermentation process, surplus microbial cells are prepared so that they can be produced as powders for the production of industrial microbial preparations for this purpose. Industrial lactic acid bacteria preparations are often expensive, since the growth and development of lactic acid bacteria is slow and the preparation to maintain a viable state for a long time is difficult. The lactic acid bacteria of the present invention can produce considerable added value when used because not only growth and development in soybean meal extract solutions is active, but also formulation is not difficult.

On the other hand, with the recent increase in interest in environmental pollution, biodegradable plastics are receiving attention, of which lactic acid polymer PLA (polylactic acid) is representative. Lactic acid required for preparing PLA is mainly produced by fermentation using lactic acid bacteria, and when the extraction solution and lactic acid bacteria of the present invention are used, an economical lactic acid preparation process can be performed.

A method of preparing fermented soybean meal according to one embodiment of the present invention comprises the steps of:

a step of obtaining a soybean meal extract solution and residual soybean meal by extracting raw soybean meal with an extraction solvent, and

a step of solid culturing by inoculating a soybean meal fermenting microorganism into a fermentation raw material including one or more selected from the group consisting of raw soybean meal and residual soybean meal.

A method of preparing fermented soybean meal according to one example of the present invention will be described in detail step by step.

First, a method for preparing fermented soybean meal according to one example of the present invention includes a process of obtaining a soybean meal extract solution and residual soybean meal by extracting raw soybean meal with an extraction solvent. Specifically, it may be performed by a process of extracting raw soybean meal with an extraction solvent and treating the extract by a solid-liquid separation process to obtain a soybean meal extract solution and residual soybean meal.

The solvent used for extracting the raw soybean meal of the present invention may be water, a lower alcohol of 1 to 6 carbon atoms, or a mixed solvent thereof (alcohol aqueous solution), and the lower alcohol of 1 to 6 carbon atoms may include one or more of ethanol, methanol, propanol, butanol, and the like. The extraction solvent can be controlled under appropriate pH conditions by using a pH adjusting agent such as an acid or a base. For example, examples of the pH adjusting agent may include hydrochloric acid, acetic acid, calcium phosphate, sodium hydroxide, citric acid, and the like. The pH condition of the extraction solvent may be pH 7.5 or less, pH 7 or less, pH 6 or less, pH 5 or less, pH 4.5 or less, below pH 4.5, pH 4.4 or less, pH 4.3 or less, pH 4.2 or less, pH 4.1 or less, pH 4 or less, pH3.5 or less, pH3 or less, or pH2 or less, and may be, for example, pH2 to 7.5, pH2 to 7, pH2 to 6.5, pH2 to 6, pH2 to 5.5, pH2 to 5, pH2 to less than 4.5, pH2 to 4.4, pH2 to 4.3, pH2 to 4.2, pH2 to 4.1, pH2 to 4, pH2 to 3.5, pH2 to 3, pH2 to 2.5, pH2.5 to 7.5, pH2.5 to 7, pH2.5 to 6.5, pH2 to 5.5, pH2.5 to 4.5, pH2.5 to 5, pH2.5 to 4.5, pH2 to 5, pH2.5 to 4.5, pH2.5, pH2 to 4.5, pH 5 to 4.5, pH2.5, pH2 to 5, pH 5 to 4.5, pH2.5, pH, pH2.5 to 4.2, pH2.5 to 4.1, pH2.5 to 4, pH2.5 to 3.5, pH2.5 to 3, pH3 to 7.5, pH3 to 7, pH3 to 6.5, pH3 to 6, pH3 to 5.5, pH3 to 5, pH3 to less than 4.5, pH3 to 4.4, pH3 to 4.3, pH3 to 4.2, pH3 to 4.1, pH3 to 4, pH3 to 3.5, pH3.5 to 7.5, pH3.5 to 7, pH3.5 to 6.5, pH3.5 to 6, pH3.5 to 5.5, pH3.5 to 5, pH3.5 to less than 4.5, pH3.5 to 4.4, pH3.5 to 4.3, pH3.5 to 4.2, pH3.5 to 4.1, pH3.5 to 4, pH 4 to 7.5, pH 4 to 7, pH 4 to 6.5, pH 4 to 6, pH 4 to 5.5, pH 4 to 5, pH 4 to less than 4.5, pH 4 to 4.4, pH 4 to 4.3, pH 4 to 4.2, or pH 4 to 4.1, and preferably may be pH2 to 4.5.

Preferably, the extraction solvent may be water, and the pH condition may be pH2 to less than 4.5, pH2.5 to less than 4.5, pH3 to less than 4.5, pH3.5 to less than 4.5, or pH 4 to less than 4.5, preferably pH2 to less than 4.5. When alcohol is used as the extraction solvent, an alcohol recovery apparatus is required, and there is a side effect that the residual alcohol component inhibits the growth and development of microorganisms, but when water or an aqueous hydrochloric acid solution is used as the extraction solvent, no additional recovery apparatus is required, and there is no negative influence on the growth and development of microorganisms, and thus it is more preferable.

The temperature of the extraction solvent may be adjusted to an appropriate temperature in consideration of the extraction efficiency of the soybean meal, and furthermore, when the fermenting microorganism is inoculated into the soybean meal extraction solution after the extraction step, the temperature of the extraction solvent may be controlled in consideration of the optimal growth and development conditions of the fermenting microorganism, and thus is preferably set in consideration of the extraction efficiency and the following process conditions.

For example, when water is used as the extraction solvent, the temperature of the extraction solvent may be 0 to 60 ℃,0 to 55 ℃,0 to 50 ℃,0 to less than 40 ℃,0 to 35 ℃,0 to 30 ℃,0 to 25 ℃,0 to 20 ℃,0 to 15 ℃,0 to 10 ℃,0 to 5 ℃,5 to 60 ℃,5 to 55 ℃,5 to 50 ℃,5 to less than 40 ℃,5 to 35 ℃,5 to 30 ℃,5 to 25 ℃,5 to 20 ℃,5 to 15 ℃,5 to 10 ℃, 10 to 60 ℃, 10 to 55 ℃, 10 to 50 ℃, 10 to less than 40 ℃, 10 to 35 ℃, 10 to 30 ℃, 10 to 25 ℃, 10 to 15 ℃, 15 to 60 ℃, 15 to 50 ℃, 15 to 25 ℃, 15 to 20 ℃, 20 to 60 ℃, 20 to 55 ℃, 20 to 50 ℃, 20 to less than 40 ℃ when water is used as the extraction solvent in view of the extraction efficiency of soybean meal, 20 to 35 ℃, 20 to 30 ℃, 20 to 25 ℃, 25 to 60 ℃, 25 to 55 ℃, 25 to 50 ℃, 25 to less than 40 ℃, 25 to 35 ℃, 25 to 30 ℃, 30 to 60 ℃, 30 to 55 ℃, 30 to 50 ℃, 30 to less than 40 ℃, 30 to 35 ℃, 35 to 60 ℃, 35 to 55 ℃, 35 to 50 ℃, or 35 to less than 40 ℃, preferably 20 to less than 40 ℃. When an alcohol or an alcohol aqueous solution is used as the extraction solvent, the temperature of the extraction solvent may be raised to a temperature of 50 to 70 ℃ for use. In the case of ethanol, since sugar extraction efficiency in soybean meal is low when used at room temperature, it is preferably used by heating to a temperature higher than room temperature.

In addition, in the case of considering the optimal growth and development conditions of the fermenting microorganisms, it is preferable that the temperature of the extracted soybean meal extract solution and the optimal growth and development temperature of the soybean meal fermenting microorganisms inoculated thereto are similar, and thus, when the enterococcus faecium SLB130 having the optimal growth and development temperature of 40 to 45 ℃ is used, the temperature of the extraction solvent may be adjusted to a temperature of 20 to 60 ℃ or 40 to 60 ℃ so that the temperature of the soybean meal extract solution is 40 to 45 ℃.

In the present invention, the extraction solvent of the raw soybean meal may be extracted by mixing the extraction solvents in the following weight ratios, based on the weight of the raw soybean meal: 1: 1 to 1: 10, 1: 1 to 1: 9.5, 1: 1 to 1: 9, 1: 1 to 1: 8.5, 1: 1 to 1: 8, 1: 1 to 1: 7.5, 1: 1 to 1: 7, 1: 1 to 1: 6.5, 1: 1 to 1: 6, 1: 1 to 1: 5.5, 1: 1 to 1: 5, 1: 1 to less than 1: 5, 1: 1 to 1: 4.9, 1: 1 to 1: 4.8, 1: 1 to 1: 4.7, 1: 1 to 1: 4.6, 1: 1 to 1: 4.5, 1: 1 to 1: 4.4, 1: 1 to 1: 4.3, 1: 1 to 1: 4.2, 1: 1 to 1: 4.1, 1: 1 to 1: 4 or less, more than 1: 2: 1 to 1: 5: 4.5 to 1: 4.5, more than 1: 4.5 to 1: 4.5, more than 1: 4.5 to 1: 4.5, more than 1: 4.1: 4.5, more than 1: 4.1: 4.5, Greater than 1: 2.5 to 1: 4.2, greater than 1: 2.5 to 1: 4.1, greater than 1: 2.5 to 1: 4 or less, 1: 3 to less than 1: 5, 1: 3 to 1: 4.9, 1: 3 to 1: 4.8, 1: 3 to 1: 4.7, 1: 3 to 1: 4.6, 1: 3 to 1: 4.5, 1: 3 to 1: 4.4, 1: 3 to 1: 4.3, 1: 3 to 1: 4.2, 1: 3 to 1: 4.1, 1: 3 to 1: 4 or less, greater than 1: 3 to less than 1: 5, greater than 1: 3 to 1: 4.9, greater than 1: 3 to 1: 4.8, greater than 1: 3 to 1: 4.7, greater than 1: 3 to 1: 4.6, greater than 1: 3 to 1: 4.5, 1: 3 to 1: 4.1: 4, 1: 3 to 1: 4.5, greater than 1: 3, 1: 4.5, greater than 1: 4.1: 3 to 1: 4.2, 1: 3, 1: 4.5, greater than 1: 4.5, greater than 1: 4.1: 4.6, 1: 3 to 1 1: 3.5 to 1: 4.8, 1: 3.5 to 1: 4.7, 1: 3.5 to 1: 4.6, 1: 3.5 to 1: 4.5, 1: 3.5 to 1: 4.4, 1: 3.5 to 1: 4.3, 1: 3.5 to 1: 4.2, 1: 3.5 to 1: 4.1, 1: 3.5 to 1: 4 or less, 1: 1 to 1: 4, 1: 1 to 1: 3.5, 1: 1 to 1: 3, 1: 1 to 1: 2.5, 1: 1 to 1: 2, 1: 1.5 to 1: 4.5, 1: 1.5 to 1: 4, 1: 1.5 to 1: 3.5, 1: 1.5 to 1: 3, 1: 1.5 to 1: 2.5, 1: 2 to 1: 4.5, 1: 4.5 to 1: 4.5, 1: 5 to 1: 3.5, or 1: 5 to 1: 5, and preferably from less than 1: 3.

In the present invention, when the soybean meal is extracted by using the extraction solvent, a solid-liquid separation method using a screw press or a centrifugal extractor may be performed. The screw press type has a problem of a reduction in extraction efficiency because the separation net is clogged with the lapse of operation time, and thus it is preferable to use a centrifugal extractor. When the soybean meal is extracted using a centrifugal extractor, the centrifugal force (rpm) of the centrifuge may be, for example, 300g to 1000g, 400g to 1000g, 500g to 1000g, 600g to 1000g, 700g to 1000g, more preferably 700 to 900 g.

The soybean meal extract solution obtained by the extraction process of the present invention contains a large amount of polysaccharides and a small amount of proteins, and thus can be used as a culture medium for microorganisms. Since the soybean meal extract solution contains a large amount of raffinose, stachyose and the like, a discardable resource can be recovered when the soybean meal extract solution is used for the culture or fermentation of lactic acid bacteria.

The soybean meal extract solution may have a sugar concentration of 5 to 20% and/or a protein concentration of 1 to 2% (w/w). In addition, the soybean meal extract solution may have a sugar concentration of 6 to 12 brix and/or a protein concentration of 0.5 to 3% (w/w), or a protein concentration of 0.5 to 2% (w/w).

In the present invention, the moisture content of the "residual soybean meal" may be 90% or less, 85% or less, 80% w/w% or less, 75% or less, 70% or less, 65% or less, less than 65%, 64% or less, 63% or less, 62% or less, 61% or less, 60% or less, less than 60%, 59% or less, 58% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, or 5% or less, preferably may be 70% or less, more preferably may be 66% or less, for example, may be 55% to 66%.

In the present invention, "residual soybean meal" may have the following contents of sugars or indigestible oligosaccharides: 2% (w/w) or less, 1.9% (w/w) or less, 1.8% (w/w) or less, 1.7% (w/w) or less, less than 1.7% (w/w), 1.6% (w/w) or less, 1.5% (w/w) or less, 1.4% (w/w) or less, 1.3% (w/w) or less, 1.2% (w/w) or less, 1.1% (w/w) or less, 1% (w/w) or less, 0.9% (w/w) or less, 0.8% (w/w) or less, 0.7% (w/w) or less, 0.6% (w/w) or less, 0.5% (w/w) or less, 0.4% (w/w) or less, 0.3% (w/w) or less, 0.2% (w/w) or less, Or 0.1% (w/w) or less.

Some of the sugar or indigestible oligosaccharide is removed by the extraction process of the raw soybean meal, and when the sugar concentration based on 100 wt% of the raw soybean meal represents the concentration of the sugar or indigestible oligosaccharide, it may be 99 wt% or less, 98 wt% or less, 97 wt% or less, 96 wt% or less, 95 wt% or less, 90 wt% or less, 85 wt% or less, 80 wt% or less, 75 wt% or less, 70 wt% or less, 65 wt% or less, 60 wt% or less, 55 wt% or less, 50 wt% or less, 45 wt% or less, 40 wt% or less, 35 wt% or less, 30 wt% or less, 25 wt% or less, 20 wt% or less, 15 wt% or less, 10 wt% or less, or 5 wt% or less.

In the present invention, the "residual soybean meal" may have a crude protein content of 15% (w/w) or more, 16% (w/w) or more, 17% (w/w) or more, 18% (w/w) or more, 19% (w/w) or more, 20% (w/w) or more, 21% (w/w) or more, or 22% (w/w) or more, based on a moisture content of 65% (w/w), and may be 55% (w/w) or more, 56% (w/w) or more, 57% (w/w) or more, 58% (w/w) or more, 59% (w/w) or more, 60% (w/w) or more, 61% (w/w) or more, or 61% (w/w) or more, based on the solid content.

The method for producing fermented soybean meal of the present invention may include a step of performing fermentation using a microorganism that ferments soybean meal, specifically, a step of performing solid culture by inoculating a soybean meal-fermenting microorganism into a fermentation raw material including one or more selected from the group consisting of raw soybean meal and residual soybean meal. The method for preparing fermented soybean meal according to one example of the present invention may further include the step of drying and pulverizing the fermented soybean meal obtained in the culturing step.

In the present invention, the fermentation raw material for preparing the fermented soybean meal may be residual soybean meal obtained by extracting raw soybean meal with an extraction solvent, raw soybean meal not extracted with an extraction solvent, and a mixture thereof. The raw soybean meal and residual soybean meal were the same as described above. Further, when a mixture of residual soybean meal and raw soybean meal is used, the fermentation raw material may control the crude protein content of the fermented soybean meal by appropriately adjusting the mixing ratio of the two components. For example, when raw soybean meal and residual soybean meal are mixed at 1: 1 to 1: 1.2, the final crude protein content of the fermented soybean meal may be 50% to 54% (w/w). For example, when raw soybean meal and residual soybean meal are mixed at 1: 1 to 1: 1.4, the final crude protein content of the fermented soybean meal may be 50 to 56% (w/w). For example, when raw soybean meal and residual soybean meal are mixed at 1: 1.2 to 1: 1.4, the final crude protein content of the fermented soybean meal may be 53 to 55% (w/w).

In the present invention, the "soybean-fermenting microorganism" may be a facultative anaerobic bacterium which is a strain having an ability to ferment soybean meal, and includes, for example, enterococcus strains, Weissella sp strains, Lactobacillus strains, and the like. The facultative anaerobic bacteria may be one or more selected from the group consisting of: enterococcus faecium, Enterococcus faecalis (Enterococcus faecalis), Weissella melitensis (Weissella koreensis), Pediococcus pentosaceus (Pediococcus pentosaceus), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus lactis (Lactobacillus lactis), Lactobacillus reuteri (Lactobacillus reuteri), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus casei (Lactobacillus casei), Lactobacillus fermentum (Lactobacillus fermentum), Lactobacillus bulgaricus (Lactobacillus bulgaricus) and Lactobacillus acidophilus (Lactobacillus acidophilus).

Generally, fermentation of soybean meal is carried out under aerobic conditions by forced supply of oxygen using aerobic microorganisms such as bacillus bacteria or aspergillus fungi or saccharomyces (brewers yeast/bakers yeast) yeast. Since it is difficult to uniformly and sufficiently supply oxygen in an aerobic process and the thickness of fermentation raw materials is limited due to oxygen supply problems in aerobic solid fermentation using aerobic microorganisms, a fermentation space and equipment are required. In addition, when fermentation is performed using conventional absolute anaerobic bacteria such as bifidobacterium (bifidus), since oxygen transmission should be prevented, excessive equipment is required and operational problems are caused.

In the present invention, as the soybean meal fermenting microorganism, facultative anaerobic lactic acid bacteria can be used, and particularly lactic acid bacteria that grow well in a soybean meal extract solution can be used. Therefore, the largest problem in the solid fermentation process, that is, the problem of a certain supply of oxygen is solved, and oxygen does not need to be blocked, unlike the case of using absolutely anaerobic microorganisms, and therefore, high productivity in the fermentation process can be achieved.

Therefore, the method for preparing fermented soybean meal according to one example of the present invention includes a step of performing solid culture of a fermentation raw material using microorganisms that ferment soybean meal, and solid fermentation may be performed in an open solid fermentation tank in which movement of oxygen or air is not restricted, and preferably, it may not include an aeration process for forcibly supplying or additionally supplying oxygen during the fermentation process. The solid fermentation according to the present invention may be carried out by a batch or continuous method, and a packed solid fermentation tank or a tray solid fermentation tank or the like may be used, or a conveyor solid fermentation tank that carries out solid fermentation on a conveyor belt may be used, and is not particularly limited.

For example, an open continuous solids fermentor can comprise: a fermenting chamber equipped with an inlet into which a fermenting material and/or lactic acid bacteria flow and an outlet from which a fermentation product flows and in which fermentation is performed; a transfer conveyor installed inside the fermenting chamber and conveying the fermentation raw material and/or the lactic acid bacteria from the inlet to the outlet; a blade mounted on a rotating shaft installed in a lateral direction from an inlet to a fermenting chamber and set up a height of a fermenting material in the fermenting chamber; and a scraper which is assembled to a rotary and counter-rotary shaft equipped to the fermenting chamber in a lateral direction from the outlet through an insertion bracket, scrapes off the fermented product fermented in the fermenting chamber, and discharges the fermented product by a predetermined amount.

In addition, in the solid fermentation method of the present invention, when facultative anaerobic lactic acid bacteria such as enterococcus faecium, Weissella gazensis, Lactobacillus plantarum, etc. are used, lactic acid generated during fermentation has a great advantage of lowering the pH of fermented soybean meal, thereby preventing contamination of various bacteria.

In addition, anaerobic fermentation using lactic acid bacteria generally has the disadvantage of a slow rate. In order to solve the above problems, in the present invention, it is preferable to prepare a soybean meal extract solution containing sugars and some proteins in soybean meal, and select a strain showing a high specific growth rate when it is used as a medium for microorganisms fermenting soybean meal, and select extraction and fermentation processes based on the strain.

In an embodiment in accordance with the inventionIn the case of the solid culture of example, a strain having an ability to ferment soybean meal may be inoculated so that the number of bacteria immediately after inoculation into a fermentation raw material is 10⁵To 10¹⁰CFU/g、10⁵To 10⁹CFU/g、10⁵To 10⁸CFU/g、10⁵To 10⁷CFU/g、10⁵To 10⁶CFU/g、10⁶To 10¹⁰CFU/g、10⁶To 10⁹CFU/g、10⁶To 10⁸CFU/g、10⁶To 10⁷CFU/g、10⁷To 10¹⁰CFU/g、10⁷To 10⁹CFU/g、10⁷To 10⁸CFU/g、10⁸To 10¹⁰CFU/g, or 10⁸To 10⁹CFU/g, more preferably, the strain can be inoculated so that the number of bacteria is 10⁶To 10⁹CFU/g is below.

The present solid culture can be carried out at a temperature of 25 to 55 ℃, 25 to 50 ℃, 25 to 48 ℃, 25 to 45 ℃, 25 to 44 ℃ or 25 to 43 ℃ for 12 to 48 hours, 12 to 36 hours, 12 to 24 hours, 18 to 48 hours, 18 to 42 hours, 18 to 36 hours, 18 to 30 hours or 18 to 24 hours. More preferably, the solid culture may be carried out at a temperature of 25 to 45 ℃ for 12 to 36 hours. The solid culture step may be carried out under anaerobic conditions, preferably under facultative anaerobic conditions.

According to the method for producing fermented soybean meal of the present invention, the culture of the present invention may be performed by inoculating a microorganism that ferments soybean meal into a fermentation raw material, or inoculating a soybean meal fermenting microorganism into a soybean meal extract solution for liquid culture and adding the solution to the fermentation raw material. For example, a step of inoculating and culturing a facultative anaerobic soybean meal fermenting microorganism to a soybean meal extract solution to obtain a seed culture product, and a step of inoculating the seed culture product to a culture raw material to perform the culture of the present invention may be included. Specifically, in the present invention, by using lactic acid bacteria that grow well in a soybean meal extract solution, by preparing a high-concentration lactic acid bacteria culture solution using the soybean meal extract solution and then fermenting the cultured lactic acid bacteria in the soybean meal, fermentation can be performed quickly.

According to an example of the present invention, fermented soybean meal may be prepared by adding a seed fermentation solution liquid, which is fermented by inoculating a microorganism that ferments soybean meal to a soybean meal extract solution, to a fermentation raw material. The soybean meal fermenting microorganisms may be inoculated to the soybean meal extract solution at the following ratio, based on 100 wt% of the soybean meal extract solution: 0.000001 to 10 wt%, 0.000001 to 5 wt%, 0.000001 to 1 wt%, 0.000001 to 0.1 wt%, 0.000001 to 0.01 wt%, 0.000001 to 0.001 wt%, 0.000001 to 0.0001 wt%, 0.000001 to 0.00001 wt%, 0.00001 to 10 wt%, 0.00001 to 5 wt%, 0.00001 to 1 wt%, 0.00001 to 0.1 wt%, 0.00001 to 0.01 wt%, 0.00001 to 0.001 wt%, 3 00001 to 0.0001 wt%, 0.0001 to 10 wt%, 0.0001 to 5 wt%, 0.0001 to 1 wt%, 0.0001 to 0.1 wt%, 0.0001 to 0.01 wt%, 0.0001 to 0.001 wt%, 0.001 to 10 wt%, 0.001 to 5 wt%, 0.001 to 1 wt%, 0.001 to 0.001 wt%, 0.01 to 0.01 wt%, 0.1 to 1 wt%, 0.01 to 0.1 wt%, 0.1 to 5 wt%, 0.1 to 1 wt%, 0.01 to 1 wt%, 0.1 to 5 wt%, 0.1 to 1 wt%, 0.1 to 5 wt%, 0.1 wt%, 0.01 to 5 wt%, 0.1.

In the liquid culture, the culture temperature may be set according to the optimal growth and development temperature of the soybean meal fermenting microorganism, and for example, may be cultured at a temperature of 20 to 50 ℃, 20 to 45 ℃, 25 to 50 ℃, 25 to 45 ℃, 30 to 50 ℃, 30 to 45 ℃, 35 to 50 ℃, 35 to 45 ℃, 40 to 50 ℃, or 40 to 45 ℃.

In the present invention, in order to produce a large amount of an inoculum of soybean meal fermenting microorganisms, a high concentration culture method using a soybean meal extract solution as a culture medium has been developed. For this reason, first, lactic acid bacteria showed high level growth and development in the soybean meal extract solution prepared by the method. The main condition is that the protein is composed of proteins such as Trypsin Inhibitor (TI) and β -conglycinin, which are anti-nutritional factors of soybean, and is decomposed to a level of small protein or peptide, so that livestock can smoothly digest soybean meal protein having a relatively high molecular weight, and the secondary condition is that microorganisms which can cause fermentation by adapting to the soybean meal solid fermentation conditions such as moisture content, temperature, pH, and the like. Bacteria in general, which exist in the general natural environment but not in the intestinal tract, have many beneficial points of fermentation in particular because the optimal temperature for growth and development is high, because many bacteria cannot grow at 40 ℃ or higher. Furthermore, since the body temperature is about 41 ℃ in the case of chickens or cattle, it may be a characteristic of lactic acid bacteria that are beneficial for survival and proliferation in the actual intestinal tract of livestock.

The enterococcus faecium SLB130 strain selected according to the research method of the present invention showed rapid growth and development in the soybean meal extract solution, and showed that the enterococcus faecium SLB130 strain had a high level of growth and development rate at 40 to 45 ℃, which is not comparable to bacillus subtilis or general industrial lactic acid bacteria, which are generally used for soybean meal fermentation. This is evidence that the selected enterococcus faecium SLB130 strain efficiently decomposes and utilizes soybean meal organic substances such as oligosaccharides, carbohydrates, proteins, etc., even at 40 ℃ or higher, which means, in particular, that soybean oligosaccharides such as sucrose, stachyose, raffinose, etc., which cannot be well utilized by common microorganisms other than lactic acid bacteria, can be effectively used as a carbon source. The optimum growth temperature of the enterococcus faecium SLB130 strain is 40-45 ℃.

The enterococcus faecium SLB130 strain is significantly superior in growth and development in a soybean meal extract solution, compared to other lactic acid bacteria or bacillus subtilis, and the like. In addition, the present inventors confirmed that when solid culture of soybean meal is performed using the strain, high-quality fermented soybean meal can be prepared, the digestibility and feed efficiency of the fermented soybean meal are improved, the content of anti-nutritional factors such as indigestible polysaccharides can be reduced by hydrolytic depolymerization of soybean protein and inactivation of trypsin inhibitor, and the crude protein content is controlled according to the mixing ratio of raw soybean meal and extracted residual soybean meal.

In the case of lactic acid bacteria, there are disadvantages in that the availability of soybean oligosaccharides is excellent and the activity of protease is relatively low, but it was shown that the enterococcus faecium SLB130 strain has sufficient competitiveness due to its excellent proteolytic ability. Specifically, enterococcus faecium SLB130 strain has excellent polymer peptide-decomposing ability, and therefore, it was confirmed by SDS-PAGE analysis that the average molecular weight of the protein in the fermented soybean meal was reduced, and thus when used as a feed, the digestibility was excellent (example 7).

For example, in the protein distribution of the raw soybean meal, the content of molecular weight in the range of less than 25kD may be 1 to 25 wt%, the content of molecular weight in the range of 25 to less than 50kD may be 10 to 60 wt%, and the content of molecular weight in the range of 50 or higher kD may be 30 to 80 wt%, based on 100 wt% of the total protein in the raw soybean meal.

For example, in the protein profile of fermented soybean meal in which residual soybean meal is fermented with the SLB130 strain, the content of molecular weight in the range of less than 25kD may be 60 to 99.9 wt%, 60 to 99 wt%, 60 to 98.5 wt%, or 60 to 98 wt%, the content of molecular weight in the range of 25 to less than 50kD may be 0.01 to 30 wt%, and the content of molecular weight in the range of 50 or higher kD may be 0.01 to 10 wt%, based on 100 wt% of the total protein in the fermented soybean meal.

For example, in the protein profile of fermented soybean meal of mixed soybean meal in which residual soybean meal and raw soybean meal are fermented with the SLB130 strain, the content of molecular weight of less than 25kD may be 40 to 80 wt%, the content of molecular weight ranging from 25 to less than 50kD may be 10 to 45 wt%, and the content of molecular weight ranging from 50 or higher kD may be 0.01 to 15 wt%, based on 100 wt% of total protein in the fermented soybean meal.

For example, in the protein profile of fermented soybean meal from which soybean meal is fermented with the SLB130 strain, the content of molecular weight in the range of less than 25kD may be 30 to 40 wt%, the content of molecular weight in the range of 25 to less than 50kD may be 47 to 60 wt%, and the content of molecular weight in the range of 50 or higher kD may be 10 to 20 wt%, based on 100 wt% of the total protein in the fermented soybean meal.

For example, in the protein profile of fermented soybean meal in which raw soybean meal is fermented with the SLB120 strain, the content of molecular weight in the range of less than 25kD may be 25 to 40 wt%, the content of molecular weight in the range of 25 to less than 50kD may be 30 to 50 wt%, and the content of molecular weight in the range of 50 or higher kD may be 20 to 30 wt%, based on 100 wt% of the total protein in the fermented soybean meal.

The enterococcus faecium SLB130 strain efficiently decomposes and utilizes soybean meal organic substances such as oligosaccharides, carbohydrates, proteins, etc., particularly sucrose, stachyose, raffinose, etc., which cannot be well used as a carbon source by common microorganisms. As such, it is possible to culture microorganisms by using an extract in which oligosaccharides that reduce the performance of soybean meal as an anti-nutritional factor can not only reduce the cost of preparing an inoculum for fermentation of soybean meal, but also generate new value by additionally producing industrial lactic acid bacteria powder, or lactic acid, amino acids, or the like, as a culture medium.

The enterococcus faecium SLB130 strain was confirmed to have properties suitable for soybean meal fermentation by confirming that the optimal growth and development temperature above general bacteria is 40 to 45 ℃ (example 3). Because the optimal growth and development temperature is high, the SLB130 strain has a beneficial point of fermentation, and thus contamination by various bacteria can be reduced. Furthermore, since the body temperature of chickens or cows is about 41 ℃, survival and proliferation in the actual intestinal tracts of domestic animals are facilitated.

The enterococcus faecium SLB130 strain may have one or more properties selected from the group consisting of the following (1) to (6):

(1) the optimum growth and development temperature is 40 to 45 c,

(2) the soybean fermentation product obtained by fermentation with enterococcus faecium SLB130 strain has a crude protein content of more than 49 to 80% (w/w), 50 to 80% (w/w), more than 51 to 80% (w/w), 52 to 80% (w/w), 53 to 80% (w/w), more than 53 to 80% (w/w), 55 to 80% (w/w), 56 to 80% (w/w), more than 56 to 80% (w/w), 57 to 80% (w/w), 58 to 80% (w/w), 59 to 80% (w/w), or 60 to 80% (w/w), based on 100 wt% of the fermentation product,

(3) the content of trypsin inhibitor in the fermented product of soybean meal obtained by fermentation with the enterococcus faecium SLB130 strain is less than 3.5mg/g, less than 3mg/g, 2.5mg/g or less, less than 2.1mg/g, 1.5mg/g or less, less than 1.2mg/g, 1.1mg/g or less, 1mg/g or less, 0.9mg/g or less, or 0.8mg/g or less based on 100 wt% of the fermented product,

(4) a soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain has beta-conglycinin at 10,000ppm or less, 9,000ppm or less, 8,000ppm or less, 7,000ppm or less, 6,000ppm or less, 5,000ppm or less, 4,000ppm or less, 3,000ppm or less, 2,000ppm or less, 1,000ppm or less, 900ppm or less, 800ppm or less, 700ppm or less, 650ppm or less, less than 620ppm, 600ppm or less, 500ppm or less, 450ppm or less, less than 420ppm, 400ppm or less, 300ppm or less, 200ppm or less, or 180ppm or less, based on 100% by weight of the fermentation product,

(5) the soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain has an indigestible oligosaccharide content of 0.1 w/w% or less, 0.09 w/w% or less, 0.08 w/w% or less, 0.07 w/w% or less, 0.06 w/w% or less, 0.05 w/w% or less, 0.04 w/w% or less, or 0.03 w/w% or less based on 100 wt% of the fermentation product,

(6) the soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain has a lactic acid concentration of 3 w/w% or more, more than 3.5 w/w%, more than 3.7 w/w%, 3.8w/w or more, more than 3.9 w/w%, 4 w/w% or more, 4.5 w/w% or more, 4.6 w/w% or more, or 5 w/w% or more based on 100 wt% of the fermentation product,

(7) the fermented product of soybean meal obtained by fermentation of enterococcus faecium SLB130 strain has a pepsin digestibility of 90 w/w% or more, 91 w/w% or more, 92 w/w% or more, 93 w/w% or more, 94 w/w% or more, 94.5 w/w% or more, 95 w/w% or more, 96 w/w% or more, or 97 w/w% or more,

(8) in a protein distribution of a soybean meal fermentation product obtained by fermentation with the enterococcus faecium SLB130 strain, a protein content of less than 25kD in molecular weight is 30 to 99.9 wt%, 30 to 99.5 wt%, 30 to 99 wt%, 30 to 98.5 wt%, 30 to 98 wt%, 30 to 97.5 wt%, or 30 to 97 wt%, a protein content of 25 to less than 50kD in molecular weight is 0.01 to 60 wt%, and a protein content in the molecular weight range of 50kD or higher is 0.01 to 25 wt%, based on 100 wt% of the total protein in the fermented soybean meal.

The enterococcus faecium SLB130 strain may be cultured continuously or in batches, and more preferably, continuous culture is preferable. Based on the fact that the specific growth rate of enterococcus faecium SLB130 strain is remarkably high, the continuous culture method is a method of automatically putting a soybean meal extract solution so as to maintain a certain pH without separately using sodium hydroxide or a lactic acid neutralizing agent such as calcium carbonate or the like.

Batch culture is a method of inoculating enterococcus faecium SLB130 strain to a soybean meal extract solution to perform batch culture, and when the pH of the culture solution is lowered to 5.5 or less, it is used as a seed for producing fermented soybean meal.

The method for producing fermented soybean meal according to the present invention may use liquid culture and solid culture, but preferably the culture of the present invention includes seed culture using a soybean meal extract solution and solid fermentation using a seed culture product. In order to ferment soybean meal in large quantities, large quantities of inoculum are required. In particular, in the case of solid fermentation where it is difficult to control the fermentation environment, a large amount of inoculum is required. Therefore, preferably, the present invention includes a two-step culture of liquid seed culture using a soybean meal extract solution and solid culture by inoculating the seed culture product into a fermentation raw material.

In the case of liquid culture, it is not difficult to set a uniform environment, and therefore it is easy to perform sterilization, temperature control, pH control, oxygen supply, oxygen occlusion, or the like, and thus a large amount of inoculum is not required. Typically, about 10 per ml is used⁶To 10⁷cfu (colony forming units) or inoculum size of 1 to 5 v/v%. However, in order to ferment soybean meal using lactic acid bacteria, the use of a sufficiently large amount of inoculum has various advantages. Since lactic acid bacteria that grow and develop anaerobically (or facultative anaerobically) grow and develop at a slower rate than aerobic bacteria or yeast, fungi, etc., a larger amount of inoculum is required to effectively ferment soybean meal.

The seed culture may be carried out at 20 to 45 ℃, 25 to 45 ℃, 27 to 45 ℃, 30 to 40 ℃, 30 to 38 ℃, 30 to 35 ℃, 32 to 45 ℃, 32 to 40 ℃, 32 to 38 ℃, 32 to 35 ℃, 35 to 45 ℃, or 35 to 40 ℃, more preferably at 25 to 45 ℃.

The fermenting microorganism in the seed culture may be inoculated at a ratio of 0.5 to 4.5 wt%, 0.5 to 4 wt%, 0.5 to 3.5 wt%, 0.5 to 3 wt%, 0.5 to 2.5 wt%, 0.5 to 2 wt%, 0.5 to 1.5 wt%, 0.5 to 1 wt%, 1 to 5 wt%, 1 to 4.5 wt%, 1 to 4 wt%, 1 to 3.5 wt%, 1 to 3 wt%, 1 to 2.5 wt%, 1 to 2 wt%, or 1 to 1.5 wt% based on 100 wt% of the soybean meal extract solution.

The fermented soybean meal according to one example of the present invention may have one or more properties selected from the following properties:

(1) the lower limit of the crude protein content is 30% (w/w) or more, more than 30% (w/w), 35% (w/w) or more, 38% (w/w) or more, more than 38% (w/w), 40% (w/w) or more, 45% (w/w) or more, more than 46% (w/w), 47% (w/w) or more, 48% (w/w) or more, more than 48% (w/w), 49% (w/w) or more, more than 49% (w/w), 50% (w/w) or more, more than 50% (w/w), 53% (w/w) or more, more than 53% (w/w), 55% (w/w) or more, 56% (w/w) or more, based on 100 wt% of the fermentation product, More than 56% (w/w), or 60% (w/w) or more, and the lower limit of the crude protein content is 100% (w/w) or less, less than 100% (w/w), 95% (w/w) or less, 90% (w/w) or less, 85% (w/w) or less, 80% (w/w) or less, 75% (w/w) or less, 70% (w/w) or less, 65% (w/w) or less, or 60% (w/w) or less,

(2) the upper limit of the trypsin inhibitor content is 8mg/g or less, less than 8mg/g, 7mg/g or less, 6mg/g or less, 5mg/g or less, 4.5mg/g or less, less than 4.5mg/g, 4mg/g or less, 3.5mg/g or less, less than 3.5mg/g, 3mg/g or less, 2.5mg/g or less, less than 2.1mg/g, 2mg/g or less, 1.5mg/g or less, less than 1.2mg/g, 1.1mg/g or less, less than 1.1mg/g, 1.0mg/g or less, less than 1.0mg/g, 0.9mg/g or less, or 0.8mg/g or less, the lower limit is 0mg/g or more, more than 0mg/g, 0.0001mg/g or higher, 0.001mg/g or higher, 0.01mg/g or higher, 0.1mg/g or higher, 0.5mg/g or higher, 0.8mg/g or higher, 1mg/g or higher, higher than 1mg/g, 1.1mg/g or higher, higher than 1.1mg/g, 1.2mg/g or higher, 1.5mg/g or higher, 2mg/g or higher, 2.1mg/g or higher, higher than 2.1mg/g, 2.5mg/g or higher, 3mg/g or higher, or 3.5mg/g or higher,

(3) the upper limit of the content of beta-conglycinin is 70,000ppm or less, less than 70,000ppm, 68,000ppm or less, less than 68,000ppm, 65,000ppm or less, less than 65,000ppm, 60,000ppm or less, 50,000ppm or less, 40,000ppm or less, 35,000ppm or less, less than 35,000ppm, 30,000ppm or less, 25,000ppm or less, 20,000ppm or less, 15,000ppm or less, less than 15,000ppm, 10,000ppm or less, 9,000ppm or less, 8,000ppm or less, 7,000ppm or less, 6,000ppm or less, 5,000ppm or less, 4,000ppm or less, 3,000ppm or less, 2,000ppm or less, 1,500ppm or less, less than 1,500ppm, 1,000ppm or less, 900ppm or less, 800ppm or less, 650,000 ppm or less, 640ppm or less, 600ppm or less, or more than 2,000ppm or less, or, 450ppm or less, 420ppm or less, 400ppm or less, 350ppm or less, 300ppm or less, 250ppm or less, 200ppm or less, or 180ppm or less, with a lower limit of 0ppm or more, above 0ppm, 0.0001ppm or more, 0.001ppm or more, 0.01ppm or more, 0.1ppm or more, 1ppm or more, 10ppm or more, 100ppm or more, 150ppm or more, 200ppm or more, 300ppm or more, or 400ppm or more,

(4) the upper limit of the indigestible oligosaccharide content is less than 17.0mg/g, less than 7.0mg/g, 5.0mg/g or less, 4.0mg/g or less, 3.0mg/g or less, less than 1.70 w/w%, 1.5 w/w% or less, less than 1.1 w/w%, less than 0.6 w/w%, 0.5 w/w% or less, 0.4 w/w% or less, 0.3 w/w% or less, 0.2 w/w% or less, 0.15 w/w% or less, 0.12 w/w% or less, 0.1 w/w% or less, 0.07 w/w% or less, 0.05 w/w% or less, 0.04 w% or less, or 0.03 w/w% or less, and the lower limit is 0 w/w% or more, more than 0.0 w/w% or less, based on 100 wt% of the fermentation product, 0.0001 w/w% or higher, 0.001 w/w% or higher, or 0.01 w/w% or higher,

(5) the lower limit of the lactic acid concentration is 0.5% (w/w) or more, 1% (w/w) or more, 1.5% (w/w) or more, 2% (w/w) or more, 2.5% (w/w) or more, 3% (w/w) or more, 3.5% (w/w) or more, 3.8% (w/w) or more, 3.9% (w/w) or more, 4% (w/w) or more, 4.5% (w/w) or more, 4.6% (w/w) or more, 5% (w/w) or more, or 5.1% (w/w) or more, and the upper limit is 10% (w/w) or less, 9.5% (w/w) or less, 9% (w/w) or less, 8.5% (w/w) or less, based on 100 wt% of the fermentation product, 8% (w/w) or less, 7.5% (w/w) or less, 7% (w/w) or less, 6.5% (w/w) or less, 6% (w/w) or less, 5.5% (w/w) or less, 5.1% (w/w) or less, 4.6% (w/w) or less, 4% (w/w) or less, 3.9% (w/w) or less, 3.8% (w/w) or less, 3.7% (w/w) or less, or 3.5% (w/w) or less,

(6) the soybean meal fermentation product has a pepsin digestibility of 88 w/w% or more, 89 w/w% or more, 90 w/w% or more, 91 w/w% or more, 92 w/w% or more, 93 w/w% or more, 94 w/w% or more, 94.5 w/w% or more, 95 w/w% or more, 96 w/w% or more, or 97 w/w% or more, and

(7) in the protein distribution of the soybean meal fermentation product, the protein content having a molecular weight in the range of less than 25kD is 25 to 99.9 wt%, 25 to 99.5 wt%, 25 to 99 wt%, 25 to 98.5 wt%, 25 to 98 wt%, 25 to 97.5 wt%, or 25 to 97 wt%, the protein content having a molecular weight in the range of 25 to less than 50kD is 0.01 to 60 wt%, and the protein content having a molecular weight in the range of 50kD or higher is 0.01 to 30 wt%, based on 100 wt% of the total protein in the fermented soybean meal.

The fermented soybean meal may have a moisture content of 10% (w/w).

In addition, the fermented soybean meal according to one embodiment of the present invention may have one or more properties selected from the following:

(1) a crude protein content (w/w%) of 1-fold or more, 1.04-fold or more, 1.06-fold or more, 1.08-fold or more, 1.1-fold or more, 1.15-fold or more, 1.16-fold or more, 1.2-fold or more, 1.21-fold or more, 1.25-fold or more, 1.3-fold or more, 1.35-fold or more, 1.8-fold or more, or 2-fold or more, based on 100 wt% of the fermentation product, as compared to the crude protein content (w/w%) before fermentation,

(2) a trypsin inhibitor content (mg/g) of less than 1 fold, 0.9 fold or less, 0.8 fold or less, 0.7 fold or less, 0.6 fold or less, 0.55 fold or less, 0.5 fold or less, 0.45 fold or less, 0.44 fold or less, 0.4 fold or less, 0.35 fold or less, 0.3 fold or less, 0.27 fold or less, 0.25 fold or less, 0.21 fold or less, 0.2 fold or less, 0.15 fold or less, 0.14 fold or less, 0.13 fold or less, 0.12 fold or less, 0.11 fold or less, 0.1 fold or less, 0.009 fold or less, 0.006 fold or less, or 0.003 fold or less based on 100 wt% of the fermentation product as compared to the crude protein content (w/w%) before fermentation,

(3) a beta-conglycinin content (ppm) of less than 1 times, 0.9 times or less, 0.8 times or less, 0.7 times or less, 0.6 times or less, 0.5 times or less, 0.4 times or less, 0.3 times or less, 0.21 times or less, 0.2 times or less, 0.1 times or less, 0.25 times or less, 0.22 times or less, 0.01 times or less, 0.009 times or less, 0.008 times or less, 0.007 times or less, 0.006 times or less, 0.005 times or less, 0.004 times or less, 0.003 times or less, 0.002 times or less, or 0.001 times or less, as compared to the crude protein content (w/w%) before fermentation based on 100 wt% of the fermentation product,

(4) an indigestible oligosaccharide content (w/w%) of less than 1-fold, 0.9-fold or less, 0.8-fold or less, 0.7-fold or less, 0.6-fold or less, 0.5-fold or less, 0.4-fold or less, 0.3-fold or less, 0.2-fold or less, 0.12-fold or less, 0.11-fold or less, 0.1-fold or less, 0.09-fold or less, 0.08-fold or less, 0.07-fold or less, 0.05-fold or less, 0.04-fold or less, 0.03-fold or less, 0.29-fold or less, 0.02-fold or less, 0.025-fold or less, or 0.01-fold or less based on 100 wt% of the fermentation product,

(5) a pepsin digestibility of 1-fold or more, 1.07-fold or more, 1.1-fold or more, 1.11-fold or more, 1.12-fold or more, 1.13-fold or more, 1.14-fold or more, 1.15-fold or more, 1.16-fold or more, 1.19-fold or more, 1.2-fold or more, 1.3-fold or more, 1.4-fold or more, or 1.5-fold or more based on 100 wt% of the fermentation product,

(6) a lactic acid concentration (w/w%) of 1-fold or more, 1.5-fold or more, 2-fold or more, 2.5-fold or more, or 3-fold or more based on 100 wt% of the fermentation product, and

(7) the protein content of less than 25kD in the fermentation product is 1-fold or more, 1.1-fold or more, 1.2-fold or more, 1.3-fold or more, 1.4-fold or more, 1.5-fold or more, 1.6-fold or more, 1.7-fold or more, 1.8-fold or more, 1.9-fold or more, 2-fold or more, 2.5-fold or more, 3-fold or more, 3.5-fold or more, 3.6-fold or more, 3.7-fold or more, 3.8-fold or more, 3.9-fold or more, or 4-fold or more compared to the protein content of less than 25kD in the raw soybean meal prior to fermentation.

The fermented soybean meal may have a moisture content of 10% (w/w).

It was confirmed that the fermented soybean meal of the present invention contains lactic acid at a high concentration, thereby effectively preventing proliferation of other contaminants (table 8). The soybean meal fermenting microorganisms secrete lactic acid during fermentation, which has been shown to lower the pH of the medium and thus act to prevent the proliferation of other contaminants.

According to another example of the present invention, there may be provided a method for producing fermented soybean meal, the method comprising the steps of extracting raw soybean meal to obtain a soybean meal extract solution and residual soybean meal; and a fermentation step of inoculating and culturing a microorganism that ferments the soybean meal or a culture product of a soybean meal-fermenting microorganism to a fermentation raw material containing residual soybean meal or a mixture of the residual soybean meal and raw soybean meal, and controlling a crude protein content of the fermented soybean meal by adjusting a content of the residual soybean meal contained in the fermentation raw material in the fermentation step.

The fermentation feedstock may be selected from the group consisting of (1): raw soybean meal, (2): residual soybean meal after extraction and (3): (1) and (2) mixtures of (a) and (b). Since the protein is concentrated and the residual soybean meal obtained after extraction has a high crude protein content, the crude protein content of the fermented soybean meal increases with the increase in the content of the residual soybean meal in the case of the mixed fermentation raw material of (3), and therefore, the final crude protein content of the fermented soybean meal can be appropriately controlled by adjusting the mixing ratio of the raw soybean meal and the residual soybean meal.

For example, for solid culture, only (2) the residual soybean meal after extraction is suitable as a fermentation raw material for producing a crude protein product at a high concentration, and it can be used as a substitute for fish meal used for hatching fish (e.g., shrimp or eel, flatfish, abalone) feed. For example, for solid culture, only (1) raw soybean meal is suitable as a fermentation raw material for adult livestock such as sows and growing pigs, broilers, laying hens and the like.

For example, for solid culture, (1) raw soybean meal and (2) residual soybean meal after extraction mixed are suitable for young animals such as piglets because of high crude protein content.

In mixing the mixture of (1) raw soybean meal and (2) extracted residual soybean meal, the extracted raw soybean meal and the extracted residual soybean meal may be mixed in the following weight ratio: 1: 10 to 10: 1, 1: 9 to 9: 1, 1: 8 to 8: 1, 1: 7 to 7: 1, 1: 6 to 6: 1, 1: 5 to 5: 1, 1: 4 to 4: 1, 1: 3 to 3: 1, 1: 2 to 2: 1, 1: 1.8 to 1.8: 1, 1: 1.6 to 1.6: 1, 1: 1.5 to 1.5: 1, 1: 1 to 1: 10, 1: 1 to 1: 9, 1: 1 to 1: 8, 1: 1 to 1: 7, 1: 1 to 1: 6, 1: 1 to 1: 5, 1: 1 to 1: 4, 1: 1 to 1: 3.1: 1 to 1: 2, or 1: 1 to 1: 5. More preferably, they may be mixed in a weight ratio of 1: 1 to 1: 1.5.

Another example of the invention is to provide a feed composition comprising fermented soybean meal. The feed composition may be used as a feed for one or more selected from the group consisting of pigs, cattle, chickens, ducks, goats, sheep, dogs and cats. In addition, the feed composition can be used as a feed composition for adult livestock, piglets or hatched fish, depending on the crude protein content.

In the feed composition for adult livestock, it is preferable that the crude protein content is about 48% to 50%, and can be prepared by mass production in a short time by using only raw soybean meal as a fermentation raw material at a high drying temperature when about 63 to 65% is measured by KOH solubility. The feed composition for adult livestock can improve economic feasibility by using only raw soybean meal as a fermentation raw material.

For a feed composition for young animals, it is preferable to improve the quality of fermented soybean meal, such as increasing the crude protein content, removing anti-nutritional factors, and increasing the digestibility of amino acids, etc., so as to be suitable for rapid growth of young animals. The feed composition for young animals may have the properties of removing anti-nutritional factors, developing epithelial cells, and increasing beneficial intestinal bacteria to promote growth of young unit animals such as piglets, hatchlings, precocious broilers, and the like. The raw soybean meal and the residual soybean meal are mixed and used together as a fermentation raw material, and may be prepared by drying at a low temperature such that it is about 65-70% when measured by KOH solubility, and has a crude protein content of 52 to 54%, preferably 53% or more, which is lower than that of feed for adult livestock.

The feed composition for hatching fish is a fish meal replacement for the feed for hatching fish and for using fermented soybean meal as feed for hatching fish, the crude protein content is important, but the removal of indigestible oligosaccharides such as raffinose or stachyose is most important. The reason is that since the intestines of fish are short and thus the digestion time is insufficient, it is preferable to remove indigestible components as much as possible in advance. For this reason, only residual soybean meal is used as a fermentation raw material, and the drying temperature is as low as possible, so that the KOH solubility is 70% or more as measured for thermal denaturation, and it can be prepared so that the crude protein content is 60% or more, preferably 60 to 65%.

The hatching fish may be one or more selected from the group consisting of: such as mackerel, flounder, halibut, trout, salmon, catfish, eel, tilapia, grouper, catfish, white leg shrimp, giant tiger shrimp, fleshy shrimp, Japanese prawn, and abalone, but not limited thereto.

The present invention proposes a technical basis which can prepare various fermented soybean meals to suit applications by first extracting aqueous sugars containing indigestible oligosaccharides using a small amount of extraction solution in the soybean meal and thereby mixing the residual soybean meal remaining after the extraction with the raw soybean meal alone or in a suitable ratio.

The results show that in the fermented soybean meal according to the present invention, since the anti-nutritional factors are sufficiently inactivated, the digestibility is increased, and the soybean protein is significantly decomposed even in a short fermentation time. Further, a method for mass-culturing lactic acid bacteria by using a soybean meal extract solution obtained as a by-product of the method has been invented, thereby producing new added value.

Advantageous effects

The present invention provides a method for producing fermented soybean meal as an anaerobic fermentation process technology with high cost competitiveness in the animal feed market, which can produce fermented soybean meal with relatively simple process equipment and low cost, thereby being an alternative to conventional aerobic fermentation processes requiring a large amount of facilities and operating costs.

The fermentation process of the present invention can be stably operated compared to the conventional lactic acid bacteria fermentation process because the fermentation time is considerably short and the possibility of contamination is low. The fermented soybean meal prepared by the process of the present invention is qualitatively competitive enough compared to fermented soybean meal produced in the conventional aerobic fermentation process. In particular, in the process of the present invention, indigestible oligosaccharides interfering with digestive absorption of livestock and significantly reducing feed efficiency of soybean meal can be excellently removed. In addition, fermented soybean meal distinguished according to livestock species and targets can be produced by using the extraction process.

In the present invention, the soybean extract solution, which can be almost discarded, is recycled as a lactic acid bacteria culture medium, so that the cost can be reduced and an environmentally friendly means without process waste is also provided. In addition, by using an additional extraction solution, products such as industrial lactic acid bacteria powder or lactic acid can generate added value. Further, a technical basis for producing various fermentation products having industrial value by fermenting a soybean extract solution using various microorganisms is proposed.

In addition, the technology of studying bacteria using an aqueous soybean meal extract solution as a microbial culture medium according to the present invention provides an innovative technical basis that can effectively recover various organic byproducts other than soybean meal, which can increase the industrial value of a fermentation process in addition to soybean meal obtained through a low-cost anaerobic process.

The fermented soybean meal according to the present invention shows sufficient inactivation of anti-nutritional factors and significant degradation of soybean protein in a short fermentation time, and thus digestion and absorption become high. In addition, a method of mass-culturing lactic acid bacteria by using the obtained soybean extract solution through processing by-products has been invented, thereby generating new added values.

The invention aims to produce soybean meal fermented by lactic acid bacteria by using lactic acid bacteria. Lactic acid bacteria are economical compared to aerobic fermentation methods using bacillus subtilis or fungi, but have problems in processes such as anaerobic conditions. To overcome this, in the present invention, a high concentration lactic acid bacterium culture solution is prepared from lactic acid bacteria that grow well in a soybean meal extract solution by using the soybean meal extract solution, and then the cultured lactic acid bacteria are inoculated into the soybean meal and fermented, thereby allowing the fermentation to proceed rapidly.

The present invention is used to improve digestibility of defatted soybean meal, which is a main raw material of livestock feed, and to reduce anti-nutritional factors, thereby increasing availability of nutrients.

The fermentation method of the present invention can be stably operated compared to the conventional lactic acid bacteria fermentation method because the fermentation time is considerably short and the possibility of contamination is low. In addition, the present invention can produce fermented soybean meal in which indigestible oligosaccharides that interfere with digestive absorption by livestock, thereby significantly reducing feed efficiency of soybean meal, are easily removed, and are distinguished according to livestock species and targets by using an extraction method.

Therefore, the digestibility of the defatted soybean meal, which is a main raw material of livestock feed, can be improved, and the anti-nutritional factors can be reduced, thereby increasing the availability of nutrients.

Drawings

FIG. 1 is a result of analyzing proteins in soybean meal before fermentation and soybean meal after fermentation by SDS-PAGE. A in FIG. 1 is the result of SDS-PAGE analysis of a protein sample of raw soybean meal before fermentation, B in FIG. 1 is the result of SDS-PAGE analysis of a protein sample of fermented soybean meal (example 4-2) in which raw soybean meal is fermented by using the SLB120 strain, C to E in FIG. 1 are the results of SDS-PAGE analysis of a protein sample of fermented soybean meal (example 5-1) in which residual soybean meal is fermented by using the SLB130 strain, F in FIG. 1 is a size marker, each line sequentially shows the sizes of 250, 150, 100, 75, 50, 37, 25, 20, 15 and 10kD from the top, and G in FIG. 1 is the result of SDS-PAGE analysis of a protein sample of fermented soybean meal (example 4-1) in which raw soybean meal is fermented by using the SLB130 strain.

Detailed Description

Hereinafter, the present invention will be described in more detail by the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.