KR101202655B1 - Manufacturing method of functional additive for feedstuff using ultra high-pressure extraction and additive for feedstuff using the same - Google Patents

Abstract

Disclosed are a method of preparing a functional feed additive which can improve the digestibility of the feed and improve the productivity of the livestock product, as well as increase the immunoglobulin in the blood, and the functional feed additive produced thereby. To this end, a protease is added to the inactive dry yeast, the inactive dry yeast is pressurized at high pressure to separate the cell wall and the cell contents, the washing solution is added to the separated yeast, and the cell contents are extracted by centrifugation to extract the cell contents. Provided are a method for preparing a functional feed additive for freeze-drying the contents and a feed additive thereby. The method for producing a functional feed additive according to the present invention is a livestock industry by improving the meat quality, breeding rate and growth rate of livestock by acting so that livestock can efficiently ingest various nutrients, enzymes and growth factors, including abundant protein of yeast This has the effect of improving the productivity.

Description

MANUFACTURING METHOD OF FUNCTIONAL ADDITIVE FOR FEEDSTUFF USING ULTRA HIGH-PRESSURE EXTRACTION AND ADDITIVE FOR FEEDSTUFF USING THE SAME}

The present invention relates to a method for preparing a functional feed additive and a feed additive produced by the same, and more particularly, to a functional feed additive which improves digestibility for feed, improves productivity of livestock products, and increases immunoglobulins in blood. It relates to a manufacturing method and a feed additive produced thereby.

 Recently, the raw material for feed has been on a steep rise, making it a major concern for the feed industry and the livestock industry. This rise in international grain prices and sea freight rates is a major blow to grain importers around the world, affecting consumers in the US, Canada, and Australia, which are exporters of grains, leading to instability in grain supply and feed prices worldwide. Is causing an increase. In Korea, raw material costs account for 80% of the total cost of formulated feed, most raw materials depend on imports, and feed costs account for 40-60% of livestock production. The same surge in imported raw material prices will lead to higher feed costs and higher livestock production costs.

Accordingly, in the livestock industry, production cost reduction, environmental pollution reduction problem, and productivity improvement are presently urgent and important tasks facing livestock industry, and various feed additives have been used as one of the solutions for the livestock industry. That is, since animals do not have enough secretion of various digestive enzymes for digestion of carbohydrates, fats and proteins, studies on the use of various enzymes have been attempted to increase the digestibility of nutrients.

As such, antibiotics have been widely used as feed additives for the purpose of promoting livestock growth, improving feed efficiency and improving the quality of products. Inhibiting the growth of pathogens by adding antibiotics to the feed could promote the growth of livestock products and improve the productivity of livestock products.

Recently, however, many problems have been raised in the use of antibiotics. In particular, some countries including the European Union (EU) regulate the importation of antibiotic-fed livestock and restrict the use of antibiotics. For this reason, non-antibiotic biological agents are being developed, such as enzymes, probiotics, yeast cultures and natural antibiotics.

Yeast has an excellent amino acid composition and has an amino acid digestibility of 98% for livestock. In addition, because it converts minerals such as zinc (Zn), iron (Fe), etc. necessary for the growth of yeast into an organic state, it is an excellent mineral source and a good source of vitamins and makes good conditions for livestock nutrition and intestinal microbial growth. . And the cell wall of yeast accounts for about 30% of the total weight and is composed of high-molecular compounds such as -glucans and -mannans, so that a large amount of enzymes for their autolysis are produced and secreted.

In addition, it acts as a source of concentrated nutrients to the intestinal microorganisms, and has a strong affinity with oxygen, thus eliminating the oxygen in the intestine, making it anaerobic, and promoting the growth of anaerobic beneficial bacteria. In order to improve weight gain and feed efficiency.

Such yeast can be classified into two types specifically.

The first is in the form of a probiotic, which feeds live live yeast with a broth to a livestock and feeds it to the livestock to produce various types of enzymes and nutrients that can be produced when the yeast breeds in the intestine of the livestock. It is used as a feed additive under the assumption that it can be supplied to a cow.

Second, in the form of dry yeast, which is mainly a method of drying yeast separated after alcoholic fermentation and adding it to a feed, it is used as a feed additive for the purpose of supplying a large amount of protein contained in the yeast to livestock.

However, in the application of feed additives using conventional yeast, the thick cell wall, which is a morphological feature of yeast, has been a major obstacle to the efficient use of such yeast additives as feed. Generally, the cell wall thickness of yeast is about 0.1 to 0.4㎛, and the outer and inner layers are composed of mannoprotein and glucan, respectively. It is. Therefore, the membrane structure of the yeast has become the biggest obstacle to the livestock can not easily consume the internal material of the yeast, and has reduced the efficiency of feed additives such as probiotics or dry yeast.

In order to solve this problem, Korean Patent Laid-Open Publication No. 1995-009943 (published on July 15, 1994) discloses a variety of nutrients, enzymes, and growth factors, including protein, that yeasts can use to efficiently consume livestock. A method for producing yeast that is in the process of digestion is disclosed.

However, the method of preparing the feed additive increases the time required to destroy the cell wall according to the enzyme, there is a problem that the content of nitrogen and crude protein contained in the cell content is increased because water is used as a pH regulator.

Accordingly, a first object of the present invention is a method for producing a functional feed additive capable of smoothly extracting the cell contents of yeast from the yeast cell wall, which can help the nutrition of livestock and the growth of intestinal microorganisms as a good source of vitamins. To provide.

In addition, a second object of the present invention is to provide a feed additive prepared by the method for producing a functional feed additive described above.

In order to achieve the first object of the present invention described above, according to an embodiment of the present invention, the step of adding a protease to the inactive dry yeast to form a mixture; Separating the yeast into cell walls and cell contents by pressurizing the inactive dry yeast to which the protease is added at 50 to 100 MPa at a temperature of 40 to 60 ° C .; Centrifuging the yeast separated into the cell wall and the cell contents to extract the cell contents; And it provides a method for producing a functional feed additive comprising the step of lyophilizing the cell contents.

In addition, in order to achieve the second object of the present invention, according to an embodiment of the present invention provides a feed additive prepared by the method for producing a functional feed additive described above.

The feed additive according to the present invention acts to enable the livestock to efficiently consume various nutrients, enzymes, and growth factors, including abundant proteins of yeast, thereby improving the quality of the livestock, improving the breeding rate, and improving the productivity of the livestock industry. It can contribute greatly.

In addition, the feed additive according to the present invention can increase the immunoglobulin present in the blood of livestock, and can reduce the content of nitrogen and crude protein contained in yeast. In addition, the feed additive according to the present invention can be used as a substitute for antibiotics, which may help to reduce the amount of antibiotics in meat.

Hereinafter, functional feed additives (hereinafter referred to as "feed additives") and feed additives prepared by using the ultra-high pressure extraction method according to the preferred embodiments of the present invention will be described in detail.

According to an embodiment of the present invention, a method of preparing a feed additive includes adding protease to an inactive dried yeast, and pressurizing the inactive dry yeast at a constant temperature to a high temperature to a cell wall and cell contents. Separating, extracting the cell contents by centrifuging the inactive dry yeast separated into the cell wall and the cell contents, and lyophilizing the cell contents.

Hereinafter, each step will be described in more detail with reference to the accompanying drawings.

1 is a flow chart showing a method of manufacturing a feed additive according to an embodiment of the present invention.

Referring to Figure 1, the first step (S10) is the step of adding a protease to inactive dry yeast.

The addition amount of the protease varies depending on the type of protease, but is not particularly limited, but it is preferable to add 300 to 600 parts by weight based on 100 parts by weight of inactive dry yeast.

When the protease is less than 300 parts by weight based on 100 parts by weight of the inactive dry yeast, the time taken to separate the cell wall and the cell contents of the inactive dry yeast is increased, 600 parts by weight When used in excess, the residual amount of protease increases after hydrolysis of the cell wall.

As the inactive dry yeast, it is preferable to use an inactive yeast culture having a protein content of 35 to 43% or a molasses yeast having a protein content of 35 to 43%, and specifically Saccharomyces cerevisiae It is good to use

The protease may be any protease, as long as it can hydrolyze the cell wall of the yeast, but preferably a protease including cellulase, lysozyme or a mixture thereof and a buffer is used. Good to do. In other words, the protease may include buffer and cellulase, buffer and lysozyme, or buffer and cellulase and lysozyme. At this time, the buffer solution using a STET buffer containing sucrose (Sucrose), Tris (Tris-HCL), ethylenediamine tetraacetic acid (EDTA), and Triton X-100 (product name of Sigma) desirable. Here, the buffer solution serves to adjust the pH so that enzymes such as cellulase and lysozyme can work smoothly. At this time, the buffer solution is preferably used to adjust the pH of the enzyme to about 7.0 to 8.0.

In addition, the buffer solution is preferably 350 to 450 parts by weight, preferably about 400 parts by weight based on 100 parts by weight of inert dry yeast. And, it is preferable to use 0.05 to 0.1 parts by weight of the cellulase and lysozyme, respectively, based on 100 parts by weight of the inactive dry yeast.

The cellulase and lysozyme is 0.05 When used in parts by weight, it is difficult to hydrolyze the cell wall, and when used in excess of 0.1 parts by weight, the manufacturing cost is excessively increased, resulting in poor economic efficiency.

Specifically, it is preferable to use a buffer solution containing 8% sucrose, 5% Triton X-100, 50 mM EDTA, 50 mM Tris pH 8.0 as the STET buffer according to the present invention. More specifically, 1 liter of STET buffer solution contains sucrose 80.0g, Triton X-100 50ml, EDTA-Na 16.8g, Tris 6.0g, 4 liter STET buffer solution is sucrose 320.0g, Triton X 200 ml -100, 67.2 g EDTA-Na, 31.5 g Tris.

The second step (S20) is a step of separating the yeast into the cell wall and the cell contents by pressing the inert dry yeast to which the proteolytic enzyme is added at a high temperature through the first step (S10).

The constant temperature is preferably selected in the range of 40 to 60 ℃, the high pressure is preferably selected in the range of 50 to 100 MPa. At this time, since the time required for the inactive dry yeast to separate into the cell wall and the cell contents by the protease under high pressure may vary depending on the type of the yeast, the time for performing this step is not particularly limited, but preferably 22 It is good to carry out for 26 to 26 hours.

 In this step, by applying a high pressure to the mixture of inert dry yeast and protease, it promotes the action of the protease to shorten the time it takes to separate the inert dry yeast into the cell wall and the cell contents, Inhibit the growth of microorganisms present.

More specifically, the STET buffer used as a protease separates the cell contents from the cell wall and the cell membrane by hydrolyzing the cell wall and the cell membrane of the inactive dry yeast.

The third step (S30) is a step of extracting the cell contents by centrifuging the yeast separated into the cell wall and the cell contents through the second step (S20).

In this step, the yeast is centrifuged at about 7,000 to 12,000 rpm for about 10 minutes through a centrifugal separator to separate the supernatant formed from the cell contents.

Between the second step (S20) and the third step (S30) may further comprise the step (S25) of adding a wash solution to the yeast separated into the cell wall and the cell contents.

This step is a step of washing the residual material of the protease remaining in the yeast, and any material may be used as the washing liquid as long as it is a material for washing the residual material of the protease used in the hydrolysis reaction of the yeast. However, it is preferable to use ethanol.

The washing solution may be used 400 to 600 parts by weight based on 100 parts by weight of inert dry yeast, preferably in a ratio of about 1: 1 with inert dry yeast and protease.

The wash solution is separated from the cell contents through a third step (S30).

The fourth step (S40) is a step of lyophilizing the cell contents separated through the third step (S30).

This step is to freeze the cell contents at a temperature of -30 to -60 ℃, and to depressurize to 0.5 to 5 × 10 ^-3 torr to remove the water contained in the cell contents.

Feed additive according to an embodiment of the present invention is produced through the above-described manufacturing method.

The feed additive does not use alcohol and phenol in the process of separating cell contents from yeast, and may replace antibiotics used as an additive in feed. In addition, the feed composition can increase the digestibility of nutrients to increase the productivity of the livestock, can be stored for a long time at room temperature, can improve the immunity of the livestock.

Hereinafter, the present invention will be described in detail with reference to the preferred embodiments. However, the following Example is for illustrating this invention concretely, It does not limit only to this.

≪ Example 1 >

1. Under normal pressure of 1 atm, Saccharomyces cerevisiae 1 kg, STET buffer solution 4 kg, Lysozyme [LYSOZYME, MPBIO, Japan] 1 g was mixed to form a mixture.

2. As shown in the photo of Figure 2, the mixture was pressurized at 50 MPa at 100 MPa for 24 hours using an ultrahigh pressure machine [TFS-50L, Dima Puretech, Korea] to the cell wall and the cell contents Separated.

3. As shown in FIG. 3, 5 kg of ethanol was added to the mixture, and then the lower layer was extracted by centrifuging the mixture at 10,000 rpm for 10 minutes using a centrifuge and then removing the supernatant.

4. As shown in the photo of Figure 4, the supernatant was removed to extract the lower layer was cooled to -50 ℃, to obtain a feed additive through a freeze-drying process to reduce the pressure to 0.5 torr.

<Example 2>

As shown in Figures 2 to 4, the feed additives were prepared under the same conditions as in Example 1, but instead of 1 g of lysozyme [LYSOZYME, MPBIO, Japan] 1g of cellulase [Cellulase, Duchefa, USA] 1 g was used.

<Example 3>

As shown in Figure 2 to 4, to prepare a feed additive under the same conditions as in Example 1, 1g of lysozyme [LYSOZYME, MPBIO, Japan] instead of 1g of lysozyme [LYSOZYME, MPBIO, Japan] of Example 1 and cellulase [ Cellulase, Duchefa, USA] 1g was used.

&Lt; Comparative Example 1 &

As shown in Figures 2 to 4, the feed additives were prepared under the same conditions as in Example 1, but 4 kg of water was used instead of 4 kg of the STET buffer solution of Example 1.

Comparative Example 2

2 to 4, feed additives were prepared under the same conditions as in Example 2, but 4 kg of water was used instead of 4 kg of the STET buffer solution of Example 2.

&Lt; Comparative Example 3 &

As shown in Figures 2 to 4, the feed additives were prepared under the same conditions as in Example 3, but 4 kg of water was used instead of 4 kg of the STET buffer solution of Example 3.

Experimental Example 1 Analysis of Protein Content of Feed Additives

The feed additives were obtained as shown in Table 1 as a result of quality analysis. At this time, the amount of analyte was used 0.4g feed additive once, and repeated experiments were performed twice for each sample.

[Table 1]

As can be seen in Table 1, the feed additive of the present invention has a nitrogen content of 3 to 4%, in contrast to Comparative Examples 1 to 3, wherein the content of nitrogen is 4 to 6% and the content of crude protein is 28 to 33%. And a crude protein content of 24 to 26% was observed.

As such, the feed additive according to the present invention can reduce the nitrogen content, which is an environmental pollutant discharged through the manure of livestock by lowering the content of nitrogen and crude protein. Given that the protein content of the existing inactive dry yeast is 35 to 43%, the feed additive can reduce the protein content by ingesting only pure nucleic acids to the livestock.

Experimental Example 2 Specification Test of Feed Additives for Weaning Pigs

72 two-way hybrids (Landrace × Yorkshire × Duroc) were reported. The body weight at the start of the test was 8.47 × 1.49 kg and the specification test was carried out for 21 days.

The design of the test was 1) basic diet (CON; basal diet), 2) feed diet with 0.025% processed yeast (H25; basal diet + 0.025% processed yeast) and 3) feed diet with 0.050% processed yeast (H50; basal diet + 0.050% processed yeast) for 3 treatments, 4 replicates per treatment and 6 batches per replicate. These specification tests were conducted at Deokyoung Farm, Jeonbuk.

The basic feed was free-vegetable in the form of corn-soybean-based powder formulated according to NRC (1998) requirements, and water was adjusted to be freely eaten using an automatic water dispenser.

1. Daily weight gain, daily feed intake and feed efficiency

The body weight and feed intake of weaning pigs were measured for 3 weeks per week to calculate the daily gain, feed intake and feed efficiency.

The effect of dietary yeast supplementation on productivity in weaning piglets is shown in Table 2. The values of SE ² in Table 2 were analyzed by variance analysis using 'General Linear Model Procedure' of SAS (SAS Korea, 1996) software, which is a statistical analysis program, and was calculated by comparing 398 ^a and 371 ^b .

Referring to Table 2 shown below, the feed efficiency of week 1 was found to be the same as the basic feed and H25 feed, but the H50 feed was higher than the other feed.

At 2 weeks, the H25 diet was lower than the basic diet, but the H50 diet was higher than the basic diet.

Feed efficiency at 3 weeks was improved by more than 5% in H25 and H50 diets.

Feed efficiency was higher in the H25 and H50 diets than in the basal diets. In other words, even if the feed additive according to the present invention is added in a small amount of the feed content of 0.025% and 0.050% it was confirmed that the feed efficiency is improved.

[Table 2] Effects of Processing Yeast on Growth Capacity of Weaning Pigs

2. Nutrient digestibility

In order to measure the nutrient digestibility, test feed supplemented with 0.2% of chromium oxide as a test indicator was fed for 7 days before the end of the test, and manure was collected by the anal massage method. The collected manure was dried in a dryer at 60 ° C. for 72 hours, and then used for analysis after crushing with a milling mill. Chromium (Cr) mixed with feed ingredients and markers was analyzed by the method presented in the Association of Official Analytical Chemists Journal (1995).

The effect of dietary yeast supplementation on nutrient digestibility in weaning pigs is shown in Table 3. In Table 3, a significant difference of the mean value between the samples was examined using 'Ducan's multiple range test'. [Duncan, D. B. (1955) Multiple range and multiple F tests. Biometrics. 11, pp 1-14.]

Referring to Table 3 shown below, there was no significant difference in the treatment section in the dry matter content and nitrogen content (P> 0.05).

[Table 3] Effect of Feeding Processed Yeast on Nutrient Digestibility of Weaning Pigs

division(%) CON H25 H50 Dry matter
(Dry matter) 77.12 78.76 78.83 nitrogen
(Nitrogen) 76.44 78.27 80.42

(3) blood properties

Blood samples were randomly selected for each treatment, and at the end of each treatment, 2 ml of blood was collected using a K ₃ EDTA tube (Becton Dickinson Vacutainer Systems, USA) from the jugular vein (Jugular), and the turbidimeter [nephelometry method] Dade Berings, Germany] was used to analyze immunoglobulin G (IgG).

The effect of dietary yeast supplementation on IgG in blood in weaning pigs is shown in Table 3.

Referring to Table 4 shown below, it can be seen that the feed diet to which the feed additive of the present invention is added has higher IgG in blood than the basic feed diet to which the feed additive is not added. That is, it was confirmed that providing the feed to which the feed additive of the present invention is added to the livestock can improve the immunity of the livestock.

[Table 4] Effect of Feeding Processed Yeast on IgG in Blood of Weaning Pigs

CON H25 H50 IgG (mg / dL) 240.4 258.1 262.4

While the above has been described with reference to a preferred embodiment of the present invention, those skilled in the art will be able to variously modify and change the present invention without departing from the spirit and scope of the invention as set forth in the claims below. It will be appreciated.

1 is a flow chart showing a method of manufacturing a functional feed additive according to an embodiment of the present invention.

Figure 2 is a diagram showing a cell wall hydrolyzed through the Examples and Comparative Examples of the present invention.

Figure 3 is a diagram showing the cell contents extracted from yeast through Examples and Comparative Examples of the present invention.

Figure 4 is a diagram showing the cell contents lyophilized through the examples and comparative examples of the present invention.

Claims (14)

(Iii) adding proteolytic enzymes to the inactive dry yeast to form a mixture; (Ii) separating the yeast into cell walls and cell contents by pressurizing the inactive dry yeast to which the protease is added at 50 to 100 MPa at a temperature of 40 to 60 ° C .; (Iii) centrifuging the yeast separated into the cell wall and the cell contents to extract the cell contents; And (Iii) a method of producing a functional feed additive comprising lyophilizing the cell contents. The method of claim 1, The method for producing a functional feed additive, characterized in that it further comprises the step of adding a wash solution between the step (ii) and step (iii). The method of claim 2, The washing solution is a method of producing a functional feed additive, characterized in that ethanol. According to claim 1, wherein the protease is Method for producing a functional feed additive, characterized in that it comprises a cellulase and a buffer solution. According to claim 1, wherein the protease is Method for producing a functional feed additive comprising a lysozyme and a buffer solution. According to claim 1, wherein the protease is A method of producing a functional feed additive, comprising cellulase, lysozyme and a buffer solution. The method according to any one of claims 4 to 6, The buffer solution is a method for producing a functional feed additive, characterized in that the STET buffer solution. The method according to any one of claims 4 to 6, The buffer solution is a method for producing a functional feed additive, characterized in that for adding 300 to 500 parts by weight based on 100 parts by weight of inert dry yeast. The method according to any one of claims 4 to 6, The cellulase is a method for producing a functional feed additive, characterized in that the addition of 0.05 to 0.1 parts by weight based on 100 parts by weight of the inactive dry yeast. The method according to any one of claims 5 to 6, The lysozyme is a method for producing a functional feed additive, characterized in that the addition of 0.05 to 0.1 parts by weight based on 100 parts by weight of the inactive dry yeast. The method of claim 7, wherein The STET buffer solution is characterized in that 8% Sucrose, 5% Triton X-100, 50mM EDTA, and 50mM Tris pH 8.0, the method of producing a functional feed additive. The method of claim 1, wherein the inactive dry yeast is Method for producing a functional feed additive, characterized in that the inert yeast culture with a protein content of 35 to 43% by weight. The method of claim 1, wherein the inactive dry yeast is Method for producing a functional feed additive, characterized in that the molasses yeast protein content of 35 to 43% by weight. delete

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