Composition for increasing the bypass rate of proteins and method for increasing the bypass rate of proteins

Abstract

The present invention relates to a composition for increasing the rumen bypass rate of protein, which comprises a fermented product of beans fermented with fermentation bacteria including lactic acid bacteria and yeast, and which allows for concentrated digestion and absorption in the rumen of ruminants after passing through the rumen in a state where digestion and absorption in the rumen is suppressed, and a composition for reducing greenhouse gases in ruminants and a method for reducing greenhouse gases in ruminants, which can reduce the amount of methane produced in the rumen of ruminants, reduce the amount of nitrogen excreted in manure, and reduce the emission of nitrogen such as nitrous oxide and ammonia, thereby reducing greenhouse gas emissions from ruminants.

Description

[Technical Field]

[0001] The present invention relates to a composition for increasing the bypass rate and a method for increasing the bypass rate of proteins, which increases the availability of proteins and carbohydrates. [Background technology]

[0002] In the livestock industry, efforts are underway to reduce excess nitrogen through the widespread use of environmentally friendly feed in order to achieve carbon neutrality by 2050. In fact, the Feed Act is scheduled to be revised to set upper limits on crude protein in cattle feed. This revision, however, has significantly increased the importance of supplying feed ingredients with high protein availability to boost bovine productivity. For Korean beef cattle, the importance of supplying feed with high protein availability is increasing to boost weight gain and meat volume, and for dairy cows, the importance of supplying feed ingredients with high protein availability to boost milk production and increase milk protein. Furthermore, the importance of supplying feed ingredients with high protein availability that can reduce nitrogen excretion in manure is also increasing.

[0003] Ruminants utilize feed nitrogen inefficiently compared to other economic animals. For example, the nitrogen utilization efficiency for milk in dairy cows in North America and Northern Europe averages 25% and 28%, respectively. Beef cattle have even lower nitrogen utilization efficiency, with weight gain efficiency from feed nitrogen at only about 14%. Ultimately, nitrogen that is not transferred to animal tissues or milk is excreted in feces and urine, polluting soil, rivers, and air. Therefore, providing ruminants with adequate, but not excessive, dietary protein and techniques to increase their nitrogen utilization are important not only for reducing feed costs but also for reducing environmental pollution. Feed protein for ruminants can be divided into rumen degradable protein (RDP), rumen undegradable protein (RUP), and non-protein nitrogen (NPN).

[0004] Ammonia (NH3) emitted from livestock farming is a major cause of air and water pollution. Ammonia contributes to eutrophication, soil acidification, and an increase in fine dust. Ammonia is oxidized by soil microorganisms and converted into nitrous oxide (N2O), a potent greenhouse gas with a global warming impact approximately 310 times greater than that of the same amount of carbon dioxide. Summary of the Invention [Problem to be solved by the invention]

[0005] One example of the present invention is to provide a composition for increasing the rumen bypass rate of proteins, which allows digestion and absorption to occur intensively in the small intestine after passing through the rumen of a ruminant in a state where digestion and absorption in the rumen is suppressed.

[0006] Another example of the present invention is to provide a composition for reducing greenhouse gases in ruminants and a method for reducing greenhouse gases in ruminants, which can reduce methane production in the rumen of ruminants, reduce nitrogen manure excretion, and reduce emissions of nitrogen such as nitrous oxide and ammonia, thereby reducing greenhouse gas emissions from ruminants. [Means for solving the problem]

[0007] One example of the present invention relates to a composition for increasing the rumen bypass rate of protein, which comprises a fermented product of pulses fermented with fermenting bacteria including lactic acid bacteria and yeast.

[0008] Yet another embodiment of the present invention relates to a feed containing a protein composition according to an embodiment of the present invention that increases the rate of rumen bypass.

[0009] Another example of the present invention relates to a method for producing a composition for increasing the rumen bypass rate of protein, a method for producing a feed containing the composition, or a method for increasing the rumen bypass rate of protein, the method comprising the steps of fermenting beans with fermentation bacteria including lactic acid bacteria and yeast, and heat-treating the fermented beans.

[0010] Yet another example of the present invention relates to the use of a pulse fermentate fermented with fermenting bacteria including lactic acid bacteria and yeast for increasing the rumen bypass rate of proteins.

[0011] Yet another example of the present invention relates to the use of a pulse fermentation product fermented with fermenting bacteria including lactic acid bacteria and yeast for the production of a composition for increasing the rumen bypass rate of proteins.

[0012] Yet another embodiment of the present invention relates to a composition for reducing greenhouse gases in ruminants, comprising a protein composition according to an embodiment of the present invention for increasing the rate of rumen bypass.

[0013] Another embodiment of the present invention relates to a method for reducing greenhouse gas emissions in ruminants, comprising administering a protein composition according to an embodiment of the present invention that increases rumen bypass rate.

[0014] Yet another example of the present invention relates to the use of a pulse fermentate fermented with fermenting bacteria including lactic acid bacteria and yeast for reducing greenhouse gas emissions from ruminants.

[0015] The present invention will now be described in more detail. One example of the present invention relates to a composition that has a high rumen bypass rate and a high absorption rate in the small intestine, thereby improving productivity and reducing greenhouse gas emissions from ruminants. The composition according to one example of the present invention comprises a fermented product of beans fermented with a fermenting bacterium, and the fermenting bacterium may be a fermenting bacterium for forming amine groups in proteins and / or a fermenting bacterium for promoting the Maillard reaction.

[0016] One embodiment of the present invention relates to a bypass protein that passes through the rumen of a ruminant in a state where digestion and absorption are inhibited, and then is digested and absorbed primarily in the small intestine, and a feed containing the same. Specifically, fructose and other sugars in legumes are fermented using lactic acid bacteria and yeast to produce reactive sugars, and the fermented product is heat-treated to inhibit digestion and absorption in the rumen and maximize digestion and absorption in the small intestine. The bypass protein production process according to one embodiment of the present invention effectively produces bypass proteins by inducing the Maillard reaction through fermentation, which changes the components of legumes without the need for additional additives.

[0017] As used herein, "bypass" refers to passing through the rumen of a ruminant without being broken down and being broken down and absorbed in the intestine, and includes bypass fat and bypass protein products. Specifically, ruminants obtain most of their necessary amino acids from proteins that pass through the rumen, i.e., bypass proteins. However, in the case of legumes, a major feed ingredient, particularly soybean meal, which accounts for 60% of the world's plant protein feed sources, most of the protein is broken down in the rumen, with only about 25% bypassing, resulting in inefficiency. As used herein, "bypass protein" refers to protein that passes through the rumen, and "bypass rate" refers to the proportion of bypass protein in ingested protein.

[0018] In one embodiment of the present invention, the fermentation bacteria may include lactic acid bacteria and yeast. When fermenting beans using lactic acid bacteria and yeast, both species are facultative anaerobic microorganisms, which are suitable for anaerobic solid-state fermentation and allow for co-culture. Furthermore, when fermentation is performed using both lactic acid bacteria and yeast, the fermentation time can be significantly reduced by approximately half. While the fermentation time is shortened, the protein content of the fermented product is significantly increased, and the protein is cleaved to form amine groups, producing reducing sugars. Furthermore, when lactic acid bacteria and yeast are used together as fermentation bacteria, the fermented product has a higher protein content and a higher bypass rate than when fermented using lactic acid bacteria alone. Specifically, as shown in the examples of the present application, the combined use of lactic acid bacteria and yeast resulted in a high content of ruminal undegradable protein (RUP) and small intestinal absorbed protein in the fermented product, and significantly lower blood urea nitrogen concentration, methane gas production, fecal nitrogen excretion, ammonia production, and nitrous oxide production, proving the synergistic effect of the combined use of lactic acid bacteria and yeast.

[0019] Beans, such as soybean meal, contain fructose, such as stachyose and raffinose, as well as various carbohydrates, most of which are water-soluble and contain anti-nutritional factors that inhibit digestion. The present invention utilizes lactic acid bacteria and yeast that effectively metabolize anti-nutritional factors that reduce the performance of legumes as feed, regardless of their nutritional value. This means that a composition for increasing bypass rate can be provided that significantly reduces fermentation time compared to lactic acid bacteria fermentation alone and produces a final product with a high protein content.

[0020] Lactic acid bacteria are known to have particularly excellent fructose metabolism capabilities. Alpha-galactosidase (alpa-(1,6) galactosidase), an enzyme commonly found in lactic acid bacteria, breaks down fructose, such as raffinose and stachyose, which are found in large quantities in soybean meal. Raffinose and stachyose account for more than 6% of soybean meal on a dry matter basis. When a single raffinose molecule is broken down by alpha-galactosidase, it is broken down into one molecule each of three monosaccharides: galactose, fructose, and glucose. Stachyose, on the other hand, is broken down into two galactose molecules, one fructose, and one glucose molecule. Lactic acid bacteria, which secrete large amounts of alpha-galactosidase and can convert the fructose in soybean meal into a more activated form and a proportionally increased proportion of monosaccharides, can more effectively promote the Maillard reaction.

[0021] For example, the fermentation bacteria according to one embodiment of the present invention include lactic acid bacteria and yeast, and the ratio of the number of cells of the lactic acid bacteria to the number of cells of the yeast is 100:1 to 1:100, 100:1 to 1:50, 100:1 to 1:1, 50:1 to 1:50, 50:1 to 1:1, 20:1 to 1:20, 20:1 to 1:15, 20:1 to 1:10, 20:1 to 1:5, 20:1 to 1:1, 20:1 to 5:1, 20:1 to 10:1, 15:1, etc. or 1:20, 15:1 to 1:15, 15:1 to 1:10, 15:1 to 1:5, 15:1 to 1:1, 15:1 to 5:1, 15:1 to 10:1, 10:1 to 1:20, 10:1 to 1:15, 10:1 to 1:10, 10:1 to 1:5, 10:1 to 1:1, 10:1 to 5:1, 10:1 to 6:1, 10:1 to 7:1, 10:1 to 8:1, or 10:1 to 9:1.

[0022] For example, the lactic acid bacteria inoculated into the fermentation raw material may be 10 6 ~10 8 cfu / g, yeast is 10 5 ~10 7 It may be cfu / g.

[0023] The lactic acid bacteria may be one or more species selected from the group consisting of Enterococcus strains, Lactobacillus strains, Weissella strains, Leuconostoc strains, Streptococcus strains, and Lactococcus strains.

[0024] Lactic acid bacteria of the genus Enterococcus include Enterococcus faecium and Enterococcus faecalis, lactic acid bacteria of the genus Lactobacillus include Lactobacillus plantarum and Lactobacillus acidophilus, and lactic acid bacteria of the genus Weissella include Weissella koreensis and Weissella cibaria. Furthermore, lactic acid bacteria of the genus Leuconostoc include Leuconostoc citreum and Leuconostoc mesenteroides, lactic acid bacteria of the genus Streptococcus include Streptococcus thermophilus, and lactic acid bacteria of the genus Lactococcus include Lactococcus lactis.

[0025] The yeast may be one or more species selected from the group consisting of strains of the genus Saccharomyces, Torula, Xanthophyllomyces, and Pichia. The Saccharomyces yeasts include Saccharomyces cerevisiae and Saccharomyces uvarum, the Xanthophyllomyces yeasts include Xanthophyllomyces dendrorhous, the Pichia yeasts include Pichia farinosa, and the Torula yeasts include Cyberlindnera jadinii.

[0026] The fermented product contained in a composition according to one example of the present invention may be fermented with a culture medium having a moisture content of 30 to 60% by weight, 30 to 50% by weight, 30 to 45% by weight, 30 to 40% by weight, 35 to 60% by weight, 35 to 50% by weight, 35 to 45% by weight, 35 to 40% by weight, 40 to 60% by weight, 40 to 50% by weight, or 40 to 45% by weight.

[0027] The fermented product contained in a composition according to one example of the present invention may be fermented at a temperature of 20 to 40°C, 20 to 35°C, 20 to 30°C, 25 to 40°C, 25 to 35°C, 25 to 30°C, 30 to 40°C, 30 to 35°C, or 35 to 40°C.

[0028] The fermented product may have a high content of rumen undegradable protein (RUP) and / or a high content of protein absorbed in the small intestine. Thus, the composition according to one embodiment of the present invention may be a composition for increasing rumen bypass of protein and / or a composition for increasing small intestinal absorption of protein.

[0029] The crude protein content of the fermented product may be 48.5% by weight or more, 49% by weight or more, 50% by weight or more, or 51% by weight or more. The crude protein content may be the crude protein content based on a moisture content of 8% by weight.

[0030] The rumen-undegradable protein refers to a protein that is not degraded in the rumen, and may be, for example, a protein that is not degraded for 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours after treatment with rumen fluid. The rumen-undegradable protein (RUP) content of the fermentate may be 74 wt% or more, 75 wt% or more, 76 wt% or more, 77 wt% or more, 78 wt% or more, 79 wt% or more, or 80 wt% or more, based on the total crude protein content of the fermentate (100 wt%).

[0031] The small intestine absorbed protein or small intestine degraded protein is a protein that is not degraded in the rumen but is degraded and absorbed in the small intestine, for example, a protein that is digested after 1 hour of treatment with pepsin and / or 18, 19, 20, 21, 22, 23, or 24 hours of treatment with a small intestine enzyme solution. The small intestine enzyme solution may contain one or more of trypsin, chymotrypsin, amylase, and lipase. The small intestine absorbed protein content of the fermented product may be 69 wt% or more, 70 wt% or more, 71 wt% or more, or 72 wt% or more based on the total crude protein content of the fermented product (100 wt%).

[0032] The fermented product may be heat-treated after fermentation. Specifically, heat treatment is necessary to form browning complexes between activated sugars and proteins produced as a result of lactic acid bacteria fermentation. Heat treatment involves the condensation of the amino groups of amino acids in the fermented product with the carbonyl groups of reducing sugars, resulting in a Mayer reaction. Application of excessive heat results in a high bypass rate but a low small intestinal digestibility, while application of insufficient heat results in a low bypass rate. Meanwhile, amino acids such as lysine, which are crucial for feed quality, are relatively sensitive to heat and are therefore likely to be destroyed during the heat treatment process. Therefore, heat treatment of the fermented product is a sensitive process that must be precisely controlled to ensure the minimum temperature and time required for the browning reaction are used. Heat treatment of the fermented product can be performed using devices such as an extruder or expeller, or a device that employs a roasting method. Roasting, the most commonly used method, primarily involves a device that directly heats soybean meal dispersed through fins installed inside a cylindrical reactor with a flame. In the examples of the present application, a roasting type heat treatment is used as an example, and a batch type rotary drum is adopted and used as a mechanical device to accurately control the temperature and treatment time.

[0033] The heat treatment may be carried out at a temperature of 70 to 150°C, 70 to 145°C, 70 to 140°C, 70 to 130°C, 80 to 150°C, 80 to 145°C, 80 to 140°C, 80 to 130°C, 90 to 150°C, 90 to 145°C, 90 to 140°C, or 90 to 130°C.

[0034] The heat treatment may be carried out after fermentation for 5 to 60 minutes, 5 to 50 minutes, 5 to 40 minutes, 5 to 35 minutes, 5 to 30 minutes, 5 to 25 minutes, 5 to 20 minutes, 5 to 15 minutes, 5 to 10 minutes, 10 to 60 minutes, 10 to 50 minutes, 10 to 40 minutes, 10 to 35 minutes, 10 to 30 minutes, 10 to 25 minutes, 10 to 20 minutes, 10 to 15 minutes, 15 to 60 minutes, 15 to 50 minutes, 15 to 40 minutes, It may be performed for 15 to 35 minutes, 15 to 30 minutes, 15 to 25 minutes, 15 to 20 minutes, 20 to 60 minutes, 20 to 50 minutes, 20 to 40 minutes, 20 to 35 minutes, 20 to 30 minutes, 20 to 25 minutes, 25 to 60 minutes, 25 to 50 minutes, 25 to 40 minutes, 25 to 35 minutes, 25 to 30 minutes, 30 to 60 minutes, 30 to 50 minutes, 30 to 40 minutes, or 30 to 35 minutes.

[0035] For example, the heat treatment may be carried out at 90 to 100° C. for 10 to 60 minutes. In the examples of the present application, the heat treatment was carried out at as low a temperature as possible to avoid problems that may occur due to excessive heat treatment.

[0036] A composition according to one embodiment of the present invention may include a fermented product produced through a fermentation process using lactic acid bacteria and yeast and a heat treatment process without the addition of any additives, such as enzymes or sugars. The process for producing the fermented product is more economical and efficient than conventional bypass soybean meal protein production processes, and can produce a product with superior efficiency and performance. To achieve this, the process involves (1) selecting lactic acid bacteria and yeast that effectively decompose the fructose and protein in soybean meal during the metabolic process, (2) establishing solid-state fermentation conditions that maximize the decomposition of fructose into activated sugar forms, and (3) a high-temperature heat treatment process that maximizes the Maillard reaction between activated sugars in soybean meal and protein. Specifically, the Maillard reaction can be promoted through fermentation using lactic acid bacteria and yeast, which effectively decompose the fructose and protein in soybean meal into activated sugar forms, such as raffinose and stachyose, during the metabolic process, thereby increasing the bypass rate of protein. The method for increasing the rumen bypass rate according to one embodiment of the present invention uses only fermentation and heat treatment without the use of additives such as enzymes or sugars, ensuring economic efficiency. The RUP (Rumen Undegradable Protein) content is 74% or more, and the digestibility and absorption rate of RUP in the small intestine is 69% or more, making the product competitive.

[0037] In one embodiment of the present invention, the legumes may include one or more selected from the group consisting of soybeans, rapeseed, palm, peas, kidney beans, corn, mung beans, adzuki beans, soybeans, lupin beans, and their oil meal. Soybean meal, a representative legume feed ingredient, is an extremely important protein source, accounting for 60% of the world's vegetable protein feed sources based on production volume. However, in cattle, most of the soybean meal is decomposed in the rumen, with only about 25-34% bypassing the rumen, making it inefficient. Cattle obtain most of their essential amino acids from microbial proteins produced during the ruminal fermentation process and proteins that pass through the rumen, i.e., bypass proteins. Various methods for processing soybean meal have been studied to increase the bypass rate in ruminants. Methods that have been primarily used include heat treatment and treatment with lignosulfonate or formaldehyde. Recently, methods of coating soybean meal with vegetable oil have also been developed. Among these, the most widely attempted process is the heat treatment of soybean meal, which utilizes the Maillard reaction, a browning reaction that occurs when the proteins and sugars in soybean meal are heated. When the carbonyl groups of sugars and amino groups of amino acids combine due to heat, the soybean meal bypasses the microbial enzymes that cause digestion in the rumen, allowing most of the digestion to occur in the small intestine. Methods include using an extruder to heat the soybean meal at high temperatures for a short period of time, or adding reactive sugars such as xylose before heat treatment. Recently, a process has been developed that accelerates the Maillard reaction by treating soybean meal with enzymes isolated from yeast and then heat treating it.

[0038] On the other hand, industrial processes have been developed to treat soybean meal with enzymes or ferment it with microorganisms to remove antinutritional factors such as trypsin inhibitors without heat. In the latter case, molds such as Aspergillus or bacteria such as Bacillus are generally used. Some companies use an economical anaerobic fermentation process using lactic acid bacteria, but lactic acid bacteria are used because they actively break down fructose (oligosaccharide), one of the antinutritional factors in soybean meal, during the metabolic process.

[0039] Therefore, the inventors have developed a process that can increase the bypass rate of proteins by promoting the Mayer reaction through lactic acid bacteria and yeast fermentation, which effectively decomposes fructose and protein in soybean meal, such as raffinose and stachyose, into activated sugars and peptides during the metabolic process.

[0040] Another embodiment of the present invention relates to a feed containing a composition for increasing the rumen bypass rate of a protein according to an embodiment of the present invention. The feed may be feed for ruminants.

[0041] As used herein, the term "ruminant" refers to an animal belonging to the order Artiodactyla that has a rumen, and may include, for example, one or more species selected from the group consisting of Bovidae, Camelidae, Cervidae, Deeridae, and Giraffidae.

[0042] The bovine family may include, for example, one or more species selected from the group consisting of Korean ox, dairy cow, yak, bison, buffalo, impala, gaur, water buffalo, goat, sheep, goat, and antelope.

[0043] The Camelidae may include, for example, one or more species selected from the group consisting of: unicamel, bicamer, llama, alpaca, guanaco, and vicuña.

[0044] The Cervidae family may include, for example, one or more species selected from the group consisting of deer, reindeer, roe deer, wood roe, elk, and moose.

[0045] The family Papilionidae may include, for example, one or more species selected from the group consisting of the lesser mouse deer and the mouse deer.

[0046] The giraffe may include, for example, one or more species selected from the group consisting of giraffes and okapis.

[0047] Another example of the present invention relates to a method for producing a composition for increasing the rumen bypass rate of protein, or a method for producing a feed containing the composition, comprising the steps of fermenting beans with fermentation bacteria including lactic acid bacteria and yeast, and heat-treating the fermented beans.

[0048] Another example of the present invention relates to a method for increasing the rumen bypass rate of protein, comprising the steps of fermenting beans with fermentation bacteria including lactic acid bacteria and yeast, and heat-treating the fermented beans.

[0049] Another example of the present invention relates to a composition for reducing greenhouse gas emissions in ruminants, comprising a fermented product of legumes fermented with fermenting bacteria including lactic acid bacteria and yeast. In the examples of this application, when ruminants were administered a fermented product obtained by fermenting legumes using a combination of lactic acid bacteria and yeast, blood urea nitrogen was reduced, methane gas and ammonia production was reduced, and fecal nitrogen excretion was reduced. Therefore, a composition according to one example of the present invention and feed containing the same are effective in reducing greenhouse gas emissions in ruminants. Therefore, another example of the present invention relates to a method for reducing greenhouse gas emissions in ruminants, comprising administering the composition to ruminants. The greenhouse gas may be methane gas and / or nitrous oxide.

[0050] The composition according to one example of the present invention may induce one or more of the following properties (1) to (6): (1) A decrease in blood urea nitrogen concentration, for example, a fasting blood urea nitrogen concentration of 90% or less, 80% or less, or 75% or less compared to a control group (the control group may be a group administered non-fermented beans or beans fermented solely with lactic acid bacteria), (2) a reduction in methane production in the rumen, e.g., a reduction in methane production of 95% or less, 90% or less, 85% or less, 80% or less, 70% or less, 60% or less, 55% or less, 50% or less, or 45% or less relative to a control group (the control group may be a group treated with non-fermented legumes, fermented legumes for a single animal, or legumes fermented solely with lactic acid bacteria); (3) a reduction in ammonia production in the rumen, e.g., ammonia concentration of 99% or less, 98% or less, 97% or less, 96% or less, 95.5% or less, 90% or less, 85% or less, 80% or less, 70% or less, 60% or less, or 50% or less relative to a control group (the control group may be a group treated with non-fermented legumes, fermented legumes for a single animal, or legumes fermented solely with lactic acid bacteria); (4) a reduction in fecal nitrogen excretion, for example, a fecal nitrogen excretion rate of 99% or less, 98% or less, 97% or less, or 96% or less relative to a control group (the control group may be a group to which the composition according to one example of the present invention is not administered); (5) An increase in retained nitrogen, for example, a residual nitrogen increase of more than 1-fold, 1.05-fold or more, or 1.1-fold or more compared to a control group (the control group may be a group to which the composition according to one example of the present invention is not administered). (6) Improving the efficiency of nitrogen utilization by the body, for example, improving the efficiency of nitrogen utilization by 1% or more, 1.5% or more, 2% or more, 2.5% or more, 3% or more, 3.01% or more, 3.02% or more, 3.03% or more, 3.04% or more, 3.05% or more, or 3.06% or more (the control group may be a group administered with feed that does not contain a composition according to one example of the present invention).

[0051] The nitrogen availability in the body may be calculated by the following mathematical formula 1. [Mathematical formula 1] Nitrogen utilization efficiency (%) = (Nitrogen utilization rate compared to the control group) / (Nitrogen intake) x 100 [Effects of the Invention]

[0052] This invention can contribute to reducing the foul odor of manure by reducing ammonia gas (NH3) and greenhouse gases such as nitrous oxide (N2O) by reducing methane in the rumen of ruminants and reducing nitrogen excretion in manure. It can also solve greenhouse gas problems by increasing the protein utilization rate in ruminants and reducing productivity and nitrogen.

[0053] The present invention provides rumen bypass protein, increases small intestinal absorption rate, reduces ammonia generated by decomposition of excess protein by ruminant microorganisms, reduces nitrogen excretion in feces and urine, minimizes the use of unnecessary protein sources, increases productivity, and provides an environmentally friendly feed (low protein, low temperature real gas feed) that will help solve national environmental problems.

[0054] A bypass rate increasing composition according to an embodiment of the present invention can reduce greenhouse gases by reducing the amount of methane produced in the rumen and the amount of nitrogen in the feces. [Example]

[0055] [Mode for carrying out the invention] The present invention will be described in more detail below with reference to the following examples, but these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0056] Example 1: Production of fermented beans using lactic acid bacteria and yeast in combination (1) Fermentation of beans using lactic acid bacteria and yeast Beans were fermented using a combination of lactic acid bacteria and yeast as fermentation bacteria, and the crude protein content of the fermented product was measured. In this example, soybean meal was used as an example of beans, and the type of lactic acid bacteria used for soybean meal fermentation is not limited, but 1.0 × 10 Enterococcus faecium or Lactococcus lactis was used in the soybean meal. 6 / g and the number of bacteria was compared 24 hours later. E. faecium was 6.5 × 10 8 / g, Lactococcus lactis is 2.5×10 7 / g, and more efficient soybean meal fermentation was possible when Enterococcus faecium strains were used. Therefore, in this example, fermentation was carried out using Enterococcus faecium as the lactic acid bacterium.

[0057] In a pre-conditioned incubator at 30°C or 35°C, 500g of seed culture (480g of water content) was added to 1000g of soybean meal (120g of water content), and fermentation was carried out for 24 hours. The bacterial cells used for fermentation were lactic acid bacteria (Enterococcus faecium KCTC13566BP) cells (10 7 cfu / g and yeast (Saccharomyces cerevisiae) 10 6 cfu / g was used.

[0058] (2) Heat treatment of fermented beans The heat treatment of the fermented material was performed using a DMD (double mixing dryer) type dryer, which has a continuous structure with double paddles installed inside the drum and a double jacket to control the temperature at a constant level and continuously feed and discharge raw materials. After heat treatment, the fermented material was treated for 10 to 60 minutes, adjusting the heat treatment time depending on the product temperature (130°C for 10 minutes; 120°C for 15 minutes; 110°C for 20 minutes; 90°C for 35 minutes). The temperature was then limited to 60°C, and the material was dried until the moisture content was 12% or less, yielding a pulse fermented product using both lactic acid bacteria and yeast (sample name: Example 1).

[0059] Comparative Example 1: Production of fermented beans using lactic acid bacteria alone The same procedure as in Example 1 (1) was carried out, except that 100 lactic acid bacteria (Enterococcus faecium KCTC13566BP) cells were used as the fermentation bacteria. 7 The resulting fermented product was heat-treated and dried in the same manner as in Example 1 (2), to obtain a legume fermented product using lactic acid bacteria alone (sample name: Comparative Example 1).

[0060] Example 2: Measurement of pH and crude protein content After fermentation was completed and before heat treatment, distilled water was added to the fermented product in an amount twice the weight of the fermented product and stirred for 5 minutes. The pH was then measured and the crude protein content was measured using the Kjeldahl method. The moisture content of the sample was corrected to 8% by weight and the results are shown in Table 1.

[0061] [Table 1]

[0062] As shown in Table 1, it was confirmed that lactic acid was produced by lactic acid bacteria fermentation, lowering the pH of the culture medium, and the crude protein content of the fermented product of Example 1, in which lactic acid bacteria and yeast were used in combination, was approximately 3% higher than that of Comparative Example 1, in which lactic acid bacteria were also used in combination.

[0063] Example 3: Measurement of RUP and small intestinal absorbed protein content For each sample, an in vitro analysis using a protease solution was performed to analyze the rumen undigested protein (RUP). Small intestinal absorbed protein was analyzed by digestion for 1 hour using pepsin (pH 2) and for 24 hours using an enzyme solution (pH 9.0) containing trypsin, chymotrypsin, amylase, and lipase. The RUP content and small intestinal absorbed protein content are shown in Table 2, based on the total crude protein (CP) content of 100% (%CP).

[0064] [Table 2]

[0065] As shown in Table 2, the fermented product of Example 1, which was fermented using a combination of lactic acid bacteria and yeast, showed an increase in RUP content of about 10% and an increase in small intestinal absorption protein of about 4% after heat treatment. Therefore, it was confirmed that the fermented product of Example 1, which was fermented using a combination of lactic acid bacteria and yeast, was not decomposed in the rumen of ruminants, resulting in an increased bypass rate and a high small intestinal absorption rate.

[0066] Example 4. Blood urea nitrogen reduction effect of bypass beans The fermented bean product of Example 1, the fermented bean product of Comparative Example 1, or soybean meal (SAJO No. 5, crude protein 46% or more, SAJODAERIM) was fed at 350 g / kg each to six animals in each group (40-45 month old Holstein steers) for six weeks, and then blood was collected via vein and analyzed for blood urea nitrogen (BUN) and total protein concentrations. The results are shown in Table 3.

[0067] [Table 3]

[0068] As shown in Table 3, the BUN concentration 3 hours after administration of the fermented bean product of Example 1 was 9.83 mg / dL, a lower value than the fasting BUN concentration in the fermented bean product of Comparative Example 1 or the soybean meal-fed group. Comparing the BUN concentration 3 hours after administration, it was 2.65 mg / dL lower (about a 21.2% decrease) than the fermented bean product of Comparative Example 1 and 5.33 mg / dL lower (about a 35.2% decrease) than the soybean meal-fed group. Therefore, the fermented bean product of one example of the present invention was highly utilized as metabolic protein, demonstrating a remarkable effect in reducing nitrogen sources excreted in feces and urine.

[0069] Example 5. Improvement of methane metabolism in bypass beans (1) Preparation of experimental materials Ruminant juice was collected from three Korean cattle with ruminal tear ducts attached, which were kept at the animal farm attached to Sunchon National University. The collected gastric juice was kept at 39°C for 30 minutes, filtered through four layers of cheese cloth to remove feed particles, and then kept at 39°C in a thermostatic water bath.

[0070] (2) Methane gas reduction effect The fermented bean product of Example 1, the fermented bean product of Comparative Example 1, fermented soybean meal for individual animals, and soybean meal were prepared. Each sample was placed in a 160mm serum bottle at a total volume of 1.0% (DM). 100ml of gastric juice and buffer solution were dispensed at a 1:3 ratio, and the bottles were capped with butyl rubber stoppers and aluminum caps. The bottles were then incubated at 100 rpm in a shaking incubator at 39°C. N2 was bubbled throughout the entire dilution and filtration process to maintain anaerobic conditions. Samples were collected at 0, 3, 6, 12, and 24 hours, with three replicates per treatment group, for a total of two runs. Fermented soybean meal for single-stomach animals, such as pigs and chickens, is a product that can be used as a substitute for fish meal by removing anti-nutritional factors from soybean meal and making it quicker and more absorbable. After fermentation, the product is dried under conditions that prevent the Mayer reaction by ensuring that the drying temperature does not exceed 60°C.

[0071] The total gas production for each sample is shown in Table 4. As shown in Table 4, the fermented soybean meal for each animal showed significantly higher gas production, while the fermented product of Example 1 showed significantly lower gas production, likely due to a decrease in the rate of protein degradation in the rumen, which inhibited ruminal microbial fermentation. In Table 4, SEM is the standard error of the mean, and a, b, and c are numbers in the same row with different subscripts indicating significant differences (p<0.05).

[0072] [Table 4]

[0073] To analyze the methane produced over time during ruminal fermentation, samples were collected using 6 ml vacutainers. Gas chromatography (Agilent Technologies HP 5890) was used. The detector was a TCD, with a Carboxen 1006PLOT capillary column 30 m x 0.53 mm (Supelco). The analytical conditions were an oven temperature of 35°C, an injection temperature of 200°C, and a detector temperature of 200°C at the outlet. N2 gas was flowed at 3 ml / min, and the amount produced over time was analyzed.

[0074] Table 5 shows the methane production during ruminal fermentation in mM / ml. The fermented product of Example 1 had significantly lower methane production after 12 and 24 hours of culture. Specifically, the bypass fermented product of Example 1 reduced methane production by 35.9% and 45.5% after 12 and 24 hours of culture, respectively, when cultured with soybean meal. The fermented product of Comparative Example 1 reduced methane production by 31.2% and 38.8% after 12 and 24 hours of culture, respectively. Fermented soybean meal for single animals reduced methane production by 18.2% after 12 hours of culture, but increased by 30.8% after 24 hours of culture. In Table 5, SEM is the standard error of the mean, and different subscripts (a, b, and c) within the same row indicate significant differences (p<0.05).

[0075] [Table 5]

[0076] (3) Ammonia concentration analysis The culture was performed as in (2) above, and ammonia concentration was analyzed by measuring the optical density (OD) at 630 nm using a spectrophotometer (manufactured by Biochrom Ltd, CB40Fj, England) according to the method of Chaney and Marbach (1962). Specifically, NH3 reacts with hypochlorite in an alkaline solution to form NH2Cl, which then reacts with phenol to produce a blue indophenol dye. The reaction can be completed within approximately 10 minutes if the catalyst sodium nitroprusside or MnSO4 is added to accelerate the reaction. The indophenol produced in this reaction is stable and serves as a good example of an analytical method via a chemical reaction. Indophenol absorbs photons at a wavelength of 630 nm.

[0077] Ammonia concentrations throughout ruminal fermentation are shown in Table 6 in mM / L. As shown in Table 6, the ammonia (NH3-N) concentration during the 24-hour incubation period was significantly higher for the fermented soybean meal for single animals (p<0.05). The bypass-fermented soybean meal of Example 1 had significantly lower ammonia concentrations, indicating that protein breakdown does not occur in the rumen and that digestion proceeds in the small intestine. In Table 6, SEM is the standard error of the mean, and a, b, and c in the same row have different subscripts indicating significant differences (p<0.05).

[0078] [Table 6]

[0079] Example 6. Improvement of nitrogen metabolism in bypass beans Eight 14.8-month-old Korean steers were housed in eight cages and divided into two groups, a control group and an experimental group, and fed either the general diet shown in Table 7 or the diet containing the fermented product of Example 1. Water was provided ad libitum at a watering hole, and roughage was provided daily at 8:00 AM and 2:00 PM. While restrained in stanchions, each steer was individually fed using a feeding basket, with the remaining amount measured using a scale after each feeding. Concentrate feed was provided ad libitum using a DeLaval automatic feeder, and individual intake was measured and recorded daily using Delpro software. The steers were weighed before morning feeding, and total feces and urine were collected at 9:00 AM each morning and recorded. 10% feces and 5% urine samples were collected and frozen. Five days later, samples from each steer were combined and analyzed, then frozen. After the 24-day experiment, a two-day washout period was conducted to verify the experimental results, and then the control group and the experimental group were switched and the same experiment was conducted.

[0080] [Table 7]

[0081] The analysis results of nitrogen intake and fecal / urine excretion are shown in Table 8. Nitrogen intake through feed was the same in both the control and experimental groups at 160 g / d, but fecal / urine nitrogen excretion was 5 g / d lower in the experimental group than in the control group, and the nitrogen utilization rate (N retention), which indicates accumulation as body protein, was also 4.9 g / d higher in the experimental group. This indicates that the bypass protein of an embodiment of the present invention improved the efficiency of nitrogen utilization by approximately 3.063%. Korean steers require approximately 953 kg of crude protein from 6 to 30 months of age, which is equivalent to 153 kg of nitrogen, 76.5 kg of nitrous oxide, and 23.7 tons of carbon dioxide. 3.063% of 23.7 tons is 726 kg, and by increasing the efficiency of nitrogen utilization in the body by the bypass protein according to an embodiment of the present invention, it has the effect of reducing CO2 by approximately 726 kg per head of Korean beef steers up to the time of slaughter at 30 months of age. If applied to the 1.27 million Korean beef steers currently being raised in Korea, it has the effect of reducing CO2 by 922,000 tons.

[0082] Therefore, the bypass protein according to one embodiment of the present invention has been confirmed to have beneficial effects on eco-friendly livestock farming by reducing the nitrogen content excreted in feces and urine, and reducing the generation of ammonia gas and nitrous oxide gas, and to increase the nitrogen utilization rate of ruminants, thereby improving nitrogen utilization efficiency and reducing the environmental burden.

[0083] [Table 8]

Claims

1. A composition for increasing the rumen bypass rate of protein, comprising a fermented product of soybean meal fermented with fermenting bacteria including lactic acid bacteria and yeast, The yeast is used to form amine groups in proteins contained in the soybean meal and to promote the Maillard reaction of the soybean meal, The lactic acid bacterium is Enterococcus faecium, the yeast is Saccharomyces cerevisiae; the fermentate comprises amino acids and reducing sugars; the fermented product bypasses the rumen by combining the amine group of the amino acid with the carbonyl group of the reducing sugar; The fermented product is heat-treated after fermentation, The crude protein content of the fermented product is 50% by weight or more, The composition has a rumen undegradable protein (RUP) content of 74% by weight or more based on the total crude protein content of the fermented product (100% by weight).

2. The composition according to claim 1 , wherein the lactic acid bacteria are used to form amine groups in proteins contained in the soybean meal and / or to promote the Maillard reaction of the soybean meal.

3. The composition according to claim 1, which is a composition for increasing small intestinal protein absorption, and the small intestinal absorption protein content of the fermented product is 69% by weight or more based on 100% by weight of the total crude protein content of the fermented product.

4. The composition according to claim 1, wherein the fermented product is heat-treated at a temperature of 80 to 150°C after fermentation.

5. The composition according to claim 1, wherein the fermented product is heat-treated for 5 to 60 minutes after fermentation.

6. The composition of claim 1, wherein the fermented product is fermented at a temperature of 20 to 40°C.

7. The composition according to claim 1, further comprising a fermented product of one or more selected from the group consisting of rapeseed, palm, pea, kidney bean, corn, mung bean, adzuki bean, soybean, oil meal thereof, and soybean, fermented with the fermenting bacterium.

8. A feed comprising the composition of any one of claims 1 to 7.

9. The feed of claim 8, wherein the feed is a ruminant feed.

10. The feed according to claim 9, wherein the ruminant comprises one or more species selected from the group consisting of Bovidae, Camelidae, Cervidae, Deeridae, and Giraffidae.

11. Fermenting soybean meal with fermentation bacteria including lactic acid bacteria and yeast; and heat treating the fermented soybean meal. A method for producing a composition for increasing the rumen bypass rate of the protein according to any one of claims 1 to 7.

12. A step of fermenting soybean meal with fermentation bacteria including lactic acid bacteria and yeast to obtain a fermented product; and heat treating the fermented product; The yeast is used to form amine groups in proteins contained in the soybean meal and to promote the Maillard reaction of the soybean meal, The lactic acid bacterium is Enterococcus faecium, the yeast is Saccharomyces cerevisiae; the fermentate comprises amino acids and reducing sugars; the fermented product bypasses the rumen by combining the amine group of the amino acid with the carbonyl group of the reducing sugar; The fermented product is heat-treated after fermentation, The crude protein content of the fermented product is 50% by weight or more, The method for increasing the rumen bypass rate of protein, wherein the fermented product has a rumen undegradable protein (RUP) content of 74% by weight or more based on a total crude protein content of 100% by weight.

13. A composition for reducing greenhouse gas emissions from ruminants, comprising the composition according to any one of claims 1 to 7.

14. The composition according to claim 13, wherein the composition is administered to a ruminant and induces one or more of the following characteristics (1) to (6): (1) Decreased blood urea nitrogen concentration, (2) Decreased methane production in the rumen. (3) Decreased ammonia production in the rumen, (4) Reduction of nitrogen excretion in feces and urine, (5) Increased retained nitrogen, and (6) Improved efficiency of nitrogen utilization in the body.

15. 8. A method for reducing greenhouse gas emissions in ruminants, comprising administering to said ruminants a composition according to any one of claims 1 to 7.

16. 16. The method of claim 15, wherein the greenhouse gas is methane gas and / or nitrous oxide.

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