WO2014200241A1

WO2014200241A1 - Method for preparing fermented soybean meal to increase feed efficiency and milk yield of dairy cows

Info

Publication number: WO2014200241A1
Application number: PCT/KR2014/005073
Authority: WO
Inventors: 이종화; 구본탁
Original assignee: 농업회사법인 주식회사 피드업
Priority date: 2013-06-11
Filing date: 2014-06-10
Publication date: 2014-12-18
Also published as: KR101485554B1; KR20140144585A

Abstract

The present invention relates to a method for preparing a bypass protein of which the digestive absorption can intensively occur in the small intestine after passing the cow rumen while the digestive absorption thereof is suppressed. A soybean meal is (1) fermented by using lactic acid bacteria to convert fruit sugar and the like into reactive sugar; and (2) thermally treated to suppress the digestive absorption thereof in the rumen and maximize the digestive absorption thereof in the small intestine. According to method for preparing a bypass protein, the bypass protein can be effectively produced by changing ingredients of the soybean meal through fermentation without a separate additive to induce a Maillard reaction well.

Description

Method of manufacturing fermented soybean meal to increase feed efficiency and yield

The present invention relates to a fermentation method of soybean meal to increase the milk yield and feed efficiency of the cow. Soybean meal is fermented using lactic acid bacteria and then heat-treated at high temperature, and the activated sugar and protein are combined by browning reaction.

The cattle obtain most of the amino acids needed from the microbial proteins produced during the rumen fermentation or the proteins that have passed through the rumen, ie bypass proteins.

Soybean meal has the disadvantage of being inefficient because it is the most important protein source, which accounts for 60% of the world's vegetable protein feedstock by production, and most cattle are degraded in the rumen and only 25-34% are bypassed. .

Various methods of processing soybean meal have been studied in order to increase the bypass ratio in ruminants, and heat treatment or treatment with lignosulfonate or formaldehyde is mainly used. Recently, coating with vegetable oil has been developed. have.

Among them, the process of heat-treating soybean meal is the most attempted, which uses a Maillard reaction (Meylar reaction) in which proteins and sugars in soybean meal are heated during heating. In other words, when the carbonyl group of sugar (amino group) of amino acid (amino group) of sugar is combined by heat and bypasses the attack of microbial enzymes that cause digestion in the rumen, most digestion occurs in the small intestine.

Using extruder for a short time at high temperature or adding a reactive sugar such as xylose and heat treatment, etc. Recently, soybean meal using enzymes isolated from yeast The process of promoting the Maillard reaction by heat treatment after heat treatment was also developed.

On the other hand, in order to remove anti-nutritional factors such as trypsin inhibitors by non-heating methods, processes for treating soybean meal with enzymes or fermenting with microorganisms have been industrialized. In the latter case, fungi such as Aspergillus or Bacillus It is common to use such bacteria. In some cases, an anaerobic process using lactic acid bacteria is used to ferment economically. Lactic acid bacteria are known to actively break down oligosaccharides, one of the anti-nutritive factors of soybean meal, during metabolism.

Accordingly, the present inventors have developed a process to increase the ratio of bypass proteins by promoting the Maillard reaction through lactic acid bacteria fermentation, which effectively decomposes fructose in soybean meal such as Raffinose and Stachyose in metabolic processes to form an activated sugar. The processed soybean meal obtained as a result of the present invention can be secured economically by using only the fermentation process and heat treatment without additives such as enzymes or sugars. The product has a RUP (Rumen Undegradable Protein) of 65% or more and a digestive absorption rate of 90% It was found to be competitive.

An object of the present invention is to produce a bypass protein by using soybean meal, specifically to provide a soybean meal bypass protein through the lactic acid bacteria fermentation process and high temperature heat treatment process does not add any additives such as enzymes or sugars from the outside It is for. Soybean meal bypass protein of the present invention is to provide a process that can produce a product that is economical and efficient compared to the conventional soybean meal bypass protein manufacturing process in terms of efficiency and performance. To this end, (1) select lactic acid bacteria that effectively break down the fructose of soybean meal in metabolism, (2) establish solid fermentation conditions in which these fructose are degraded to the form of activated sugars, and (3) activate soybean meal. It is to develop a high temperature heat treatment process to maximize the Maillard reaction between sugar and protein.

The present invention includes the following process development steps to achieve the above object. (1) lactic acid bacteria selection step of producing an extract containing soybean meal fructose and the medium as a medium, the lactic acid bacteria excellent growth rate, (2) the decomposition of fructose in soybean meal solid phase fermentation using the lactic acid bacteria to the maximum And (3) a heat treatment step for establishing a heat treatment condition for effecting the Maillard reaction.

The present invention enables economic production of high quality bypass proteins by establishing fermentation and heat treatment conditions using lactic acid bacteria that can promote Maillard reaction by effectively metabolizing water-soluble carbohydrates, especially fructose in soybean meal.

In the present invention, anaerobic fermentation using lactic acid bacteria converts fructose and carbohydrates of soybean meal into activated sugar form by Maillard reaction, which enables to prepare bypass protein from soybean meal without using additives from the outside. . The bypass protein production method of the present invention is a simple and economical bypass protein production method.

In the present invention, the heat treatment process is performed at a relatively low temperature, thereby providing a method of reducing digestive absorption rate in the small intestine of cattle or excessive destruction of essential amino acids such as lysine by excessive heat treatment.

Processed soybean meal prepared by the process of the present invention can ensure the quality competitiveness by ensuring that the RUP is 65% or more in the quality, the digestive absorption rate in the small intestine is 90% or more.

The present invention relates to a method for producing fermented soybean meal, comprising: extracting a medium from soybean meal to culture lactic acid bacteria; Inoculating and incubating the lactic acid bacteria in the medium extracted from the soybean meal; Fermenting soybean meal using the culture medium of the culture medium in which the lactic acid bacteria were cultured and heat-treating the fermented soybean meal.

Method for producing a bypass protein by lactic acid bacteria fermentation and heat treatment from soybean meal of the present invention will be described in detail using the following examples. The following description does not limit this invention, and the changeable structure of this invention anticipated from this is within the scope of this invention.

The process of the present invention will be described in detail step by step as follows.

1. Selection of Lactobacillus

Soybean meal contains fructose and various carbohydrates represented by stakiose and raffinose, most of which are water-soluble and digestive inhibitors. In the present invention, it focuses on using lactic acid bacteria that effectively metabolize fructose and carbohydrates, which are irrelevant to the nutrition of soybean meal but rather deteriorate as a feed. That is, in the fermentation process of lactic acid bacteria, large amounts of monosaccharides such as glucose and galactose are made from fructose and carbohydrates, and these monosaccharides utilize the principle of thermally binding with proteins or peptides more effectively. .

In particular, lactic acid bacteria are known to have excellent metabolic activity against fructose. An enzyme called alpa- (1,6) galactosidase, which is commonly found in lactic acid bacteria, is contained in soybean meal such as raffinose and starchiose. It is an enzyme that breaks down fructose. The content of raffinose and stachyose in soybean meal is over 6% by dry weight. When one raffinose molecule is decomposed by alpha galactosidase, each molecule of three monosaccharides, such as galactose, fructose, and glucose, is decomposed into one molecule. Stachyose, on the other hand, is broken down into two galactose, one fructose and one glucose molecule.

In this way, the lactic acid bacteria that can secrete a lot of alpha galaccosidase enzyme and convert the fructose in soybean meal into a more active form and increase the proportion of monosaccharides can promote the Maillard reaction more effectively.

In the present invention, in order to select such lactic acid bacteria was used a method of measuring the growth rate in soybean meal aqueous extract. First, the extraction of soybean meal using water results in the dissolution of most sugar components and water-soluble carbohydrates. When the growth rate is measured by using the extract as a medium without any additives, lactic acid bacteria having high sugar metabolism from fructose and carbohydrates of soybean meal Can be selected efficiently.

The lactic acid bacteria used for culturing soybean meal extract in the present invention is Enterococcus lactic acid bacteria, Lactobacillus (Lactobacillus) lactic acid bacteria, Bisella (Weissella) lactic acid bacteria, Leuconostoc (Leuconostoc) lactic acid bacteria, Streptococcus (Streptococcus) genus Lactic acid bacteria, Lactococcus (Lactococcus) is a lactic acid bacteria which is any one selected from the lactic acid bacteria and mixed lactic acid bacteria thereof.

The Enterococcus genus lactic acid bacteria include Enterococcus faecium, Enterococcus faecalis, and Lactobacillus genus Lactobacillus (Lactobacillus plantarum), Lactobacillus plantarum (Lactobacillus plantarum) Lactobacillus acidophilus, and the Weissella genus Lactobacillus (Weissella koreensis), Weissella ciberia (Weissella cibaria). In addition, lactic acid bacteria of the genus Leuconostoc include Leuconostoc citreum and Leuconostoc mesenteroides, and streptococcus thermophillus in the strains of Streptococcus genus. There is, Lactococcus (Lactococcus) lactic acid bacteria are Lactococcus lactis (Lactococcus lactis).

One or more lactic acid bacteria are inoculated and incubated in the extract (medium) extracted from soybean meal, and the culture medium in which the lactic acid bacteria are cultured is mixed with soybean meal to ferment soybean meal as lactic acid bacteria.

2. Soybean meal fermentation using lactic acid bacteria

Soybean meal fermentation using microorganisms is mainly used in the solid substrate fermentation (solid substrate fermentation) method. After adding an appropriate amount of water to soybean meal, the microorganisms are inoculated and fermented. In order to make fermentation well, a process such as steaming to pre-treat soybean meal with hot steam may be added. Extracting the medium from the soybean meal is extracted with extract water at a temperature of 40 ~ 80 ℃, extract so that the sugar concentration of the extracted medium (extract solution) is a concentration of 1 ~ 10 Brix.

Unlike aerobic fermentation using bacteria such as fungi, yeast or Bacillus, lactic acid bacteria culture belongs to anaerobic fermentation. This is because the former microorganisms are aerobic microorganisms that require oxygen for growth and fermentation, whereas most lactic acid bacteria are facultative anaerobes. This means that both growth and fermentation are possible in the presence or absence of oxygen. For this reason, anaerobic fermentation using lactic acid bacteria is economical fermented soybean meal production is possible because there is no precise oxygen supply device, such as the imperial chamber or energy consumption. In addition, lactic acid bacteria have the advantage that the production of contaminating bacteria by the effect of lowering the pH of the fermentation product because it generates lactic acid (lactic acid) during the fermentation process.

In the present invention, a culture solution was prepared from the soybean meal extract to make the inoculum bacterium solution, and then inoculated into the soybean meal with water to establish a process condition of fermentation at a constant temperature for a certain time. Inoculum required for stable fermentation, ie inoculation rate of culture solution is 10 ⁶ ~ 10 ⁹ cfu / g, soybean meal water content is 30-60%, fermentation temperature is 20-40 ℃, fermentation period is 2-4 days Ferment during.

3. Heat treatment of fermented soybean meal

Heat treatment is essential to make browning conjugates between activated sugars and proteins produced by lactic acid bacteria fermentation. If more heat is applied, the bypass rate is higher but the digestibility in the small intestine is lower. Is lowered. On the other hand, amino acids such as lysine are very important to the quality of the feed, and are relatively bad for heat, so they are likely to be destroyed during heat treatment. Therefore, the heat treatment of fermented soybean meal is a sensitive process that must be precisely controlled to use the minimum temperature and time required for the browning reaction.

For heat treatment of soybean meal, an apparatus such as an extruder or an expeller may be used, or a roasting method may be used. Roasting, which is the most common method, is mainly used for direct heating with a flame on soybean meal dispersed by a fin installed inside a cylindrical reactor. In the present invention, a roasting type heat treatment was used, and a batch type rotary drum was used as a mechanical device for accurate control of temperature and processing time.

As the temperature used for the heat treatment, various conditions are used from 90 ° C. to 145 ° C., but preferably 30 to 120 minutes at 90 ° C. to 98 ° C. In the present invention, in order to avoid problems that may occur due to excessive heat treatment, the heat treatment conditions are aimed at as low a temperature as possible.

Example 1 Characteristics of Selected Lactic Acid Bacteria

When the soybean meal extract was used as a medium, it was proceeded as follows to select lactic acid bacteria with excellent growth rate.

Water adjusted to pH 4.4 using hydrochloric acid was used as the extraction solvent. At this time, the temperature of the solvent was 40 ~ 80 ℃ and 5 times the solvent was added to the soybean meal by weight. After sufficiently stirring for 10 minutes, the resultant was filtered through a 50-mesh sieve to obtain an extract.

The concentration of sugar in the extract was measured using a sugar meter and the unit was Brix. 1 Brix is defined as the number of grams of sugar in 100 grams of solution. Proteins were quantified by the Bradford method and used as the mass (g) of protein contained in 100 ml extract.

Extracts to be used as a medium was a sugar concentration of 5 Brix or more, protein concentration of less than 1% was used and sterilized using a membrane filter having a pore size of 0.22㎛. The growth curves of the extracts inoculated with each microorganism were measured for absorbance at 660 nm wavelength, and specific growth rates were calculated based on the growth curves.

Table 1

Specific growth rate of microorganisms

Strain name	Specific growth rate
Enterococcus faecium	1.46
Weissella koreensis	1.04
Lactobacillus plantarum	0.61
Bacillus subtilis	0.31

As shown in Table 1, when the soybean meal extract is used as a medium, the highest specific growth rate is Enterococcus faecium strain. Not only was it superior to other lactic acid bacteria, but also superior to Bacillus genus bacteria.

Example 2: Soybean meal solid phase fermentation using Enterococcus faecium (SLB120) strain

An experiment was conducted to analyze the optimal fermentation temperature of the SLB120 strain. The soybean meal with the final 40% moisture and 10 ⁷ cfu (colony forming unit) / g SLB120 cells per gram of soybean meal was added to the incubator pre-adjusted at 20 ° C, 25 ° C, 30 ° C, 35 ° C, 40 ° C, etc. for 24 hours. After fermentation, the pH of soybean meal was measured. The more lactic acid is fermented, the more lactic acid is produced, which lowers the pH of the medium. Double distilled water was added to the fermented soybean meal by weight and the pH after stirring for 5 minutes was used as the pH of soybean meal.

TABLE 2

PH by temperature

Temperature (℃)	pH
20	5.9
25	5.5
30	5.4
35	5.6
40	5.7

As shown in the above table, fermentation occurred sufficiently at the temperature conditions between 20 ° C and 40 ° C, but the decrease in pH at 25 ° C and 30 ° C was the largest, suggesting that fermentation was the most active at this temperature range. Could.

Anaerobic fermentation using lactic acid bacteria in the actual production scale proceeds without supplying air from the outside in a closed space such as a bulk bag, so as the fermentation proceeds, the temperature of the whole soybean meal increases due to the latent heat. Unless otherwise stated, 500 kg soybean meal is fermented in closed bulk bags and rises up to 40 ° C. In the present invention, the temperature at the time of inoculation of the inoculated bacteria was set at 25 to 30 ° C.

Next, an experiment was conducted to determine the optimal fermentation time of the SLB120 strain. Soybean meal inoculated with soybean meal temperature controlled at 25 ℃ was inoculated at 10 ⁷ cfu / g per gram of soybean meal and fermented for 24 hours, 36 hours, 48 hours, 56 hours, 72 hours, respectively. pH was measured.

TABLE 3

PH by fermentation time

Fermentation time (h)	pH
24	6.1
36	5.9
48	5.7
56	5.5
72	5.4

As shown in the table above, the pH of soybean meal decreased gradually over time, and after 56 hours, the decrease decreased significantly.

In the present invention, the optimum elapsed time of soybean meal fermentation using the SLB120 strain was set to 56 to 72 hours.

Example 3: Heat Treatment of Fermented Soybean Meal

Soybean meal heat treatment in the present invention used a rotary drum type dryer has a double screw conveyor is installed inside the drum has a batch structure in which the raw material continues to circulate in the drum.

In order to determine the heat treatment conditions, first, the treatment time was set to one hour, and heat treatment was performed at each temperature. The fermented soybean meal was put into the above dryer and the internal soybean meal temperature was set to 90 ° C., 95 ° C., 100 ° C., and 105 ° C. using a diesel gas burner of 200,000 Kcal / hr. Thereafter, the temperature was limited to 60 ° C. and dried until the water content was 12% or less.

Each sample was subjected to in vitro analysis using protease solution for UIP (non-degradable protein) analysis.

The following is the results of UIP analysis of fermented soybean meal heat-treated at each temperature.

Table 4

UIP by temperature

Temperature (℃)	UIP (%)
90	60
95	65
100	72
105	81

As shown above, when treated at a temperature of 95 ° C. or more for one hour, the UIP ratio was over 65%. The higher the heat treatment temperature, the higher the UIP ratio, but if excessive Maillard reaction occurs, the digestive absorption rate in the small intestine may be lowered and the breakdown of weak amino acids such as lysine may occur. It was set in the range.

In order to determine the heat treatment time, this time, the temperature was set to 95 ° C. and heat treatment was performed for 30 minutes, 60 minutes, 90 minutes, and 120 minutes, respectively. Each sample was measured for UIP in the same manner as above, and subjected to KOH solubility test. About 0.2% of the sample was added to 0.2% KOH solution and stirred at room temperature for a period of time, followed by filtration using filter paper. The filtered supernatant was measured for protein by Kjeldahl method.

Table 5 shows the results of measuring the UIP ratio and KOH solubility for each sample.

Table 5

UIP and KOH Solubility by Heat Treatment Time

Heat treatment time (minutes)	UIP (%)	KOH solubility (%)
30	45	72.1
60	65	68.2
90	72	57.3
120	80	45.4

As shown in the table, the KOH solubility was significantly reduced in the sample heat-treated for 90 minutes or more. KOH solubility is a method for analyzing the solubility of proteins in soybean meal. A low value means that the protein has a high degree of denaturation. Therefore, it can be estimated that the digestibility of the small intestine is low.

In the present invention, the heat treatment time of the fermented soybean meal was determined to be 60 to 90 minutes in order to maintain a high digestibility in the small intestine.

Example 4: Performance Analysis as Bypass Protein

Specimen testing and in-situ experiments were performed using samples prepared according to soybean meal processing for bypass proteins in the present invention.

A sample inoculated with SLB120 lactic acid bacteria at a concentration of 10 ⁷ cfu / g or more per gram of soybean meal at 25 ° C. and then fermented for 72 hours was heat treated at 95 ° C. for 60 minutes and dried to a final 12% moisture content.

In order to measure the in-situ bypass rate, experiments were conducted using cattle equipped with pistol. Experimental animals were incubated for 2 days using cattle equipped with three or more pistol. Sample bags are loaded at the same time and taken out at the same time. The ruminal position is the ventral rumen. The bag used is polyester and the pore size is 40 ~ 60um, and the sample size / surface area ratio is about 10 mg / ㎠. Sample bags that have been incubated in the rumen of experimental animals are repeated 5 times at 1 min / rinse to correct for residual microorganisms in the sample bags. Normal soybean meal was used as a control.

The following are the performance analysis results for the sample.

Table 6

In-situ bypass rate

Sample name	In-situ bypass rate (%)
Soybean meal	35
sample	69

The bypass protein manufacturing process of the present invention has the usefulness of effectively producing the bypass protein by causing a Maillard reaction to occur by changing the composition of soybean meal through fermentation without a separate additive.

In addition, the processed soybean meal prepared by the process of the present invention is useful in securing quality competitiveness by providing a RUP of 65% or more and a digestive absorption rate of 90% or more in the small intestine in its quality.

Claims

In the manufacturing method of fermented soybean meal,

Extracting the medium from the soybean meal to culture the lactic acid bacteria;

Inoculating and incubating the lactic acid bacteria in the medium extracted from the soybean meal;

Fermenting soybean meal using the culture solution of the culture medium in which the lactic acid bacteria were cultured;

Method for producing a fermented soybean meal for ruminant animals, characterized in that the fermented soybean meal heat treatment.
The method of claim 1,

Extracting the medium from the soybean meal is extracted with extract water of 40 ~ 80 ℃ temperature, the method of producing a fermented soybean meal for ruminant animal, characterized in that the sugar concentration of the extract is extracted at a concentration of 1 ~ 10 Brix.
The method according to claim 1 or 2,

The lactic acid bacteria inoculated in the extracted medium are Enterococcus lactic acid bacteria, Lactobacillus lactic acid bacteria, Weissella lactic acid bacteria, Leukonostoc lactic acid bacteria, Streptococcus (Streptococcus) lactic acid bacteria, A method for producing fermented soybean meal for ruminant animals, characterized in that it is selected from one or two or more lactic acid bacteria of Lactococcus genus lactic acid bacteria.
The method according to claim 1 or 2,

When the soybean meal is fermented using the culture solution, soybean meal is a method of producing a fermented soybean meal for ruminant animals, characterized in that the culture for 2 to 4 days at water content 30 ~ 60%, temperature 20 ~ 40 ℃.
The method according to claim 1 or 2,

The heat treatment temperature in the step of heat-treating the fermented soybean meal is a method of producing a fermented soybean meal for ruminant, characterized in that the heat treatment for 30 to 120 minutes at a temperature of 90 ~ 98 ℃.