KR101187512B1 - METHOD FOR PREPARING FEED ADDITIVE CONTAINING γ-AMINOBUTYRIC ACID - Google Patents

Abstract

PURPOSE: A producing method of a feed additive containing gamma-aminobutyric acid is provided to economically produce the feed additive by obtaining gamma-aminobutyric acid from L-glutamic acid using an enzyme reaction. CONSTITUTION: A producing method of a feed additive containing gamma-aminobutyric acid comprises the following steps: mixing absorbent excipient with 20-70%(v/w) of purified water to obtain a first mixture; mixing L-glutamic acid with the first mixture to obtain a second mixture; adding glutamic acid decarboxylase and pyridoxal phosphate into the second mixture with a pH of 4.5-5.5, and enzyme-reacting the mixture to change the L-glutamic acid into gamma-aminobutyric acid(GABA); and drying the obtained mixture of absorbent excipient and enzymatic reaction solution.

Description

Method for preparing feed additive containing gamma aminobutyric acid {Method for Preparing Feed Additive Containing γ-Aminobutyric Acid}

The present invention relates to feed additives for livestock, and more particularly, to a method for producing a feed additive containing gamma aminobutyric acid (GABA) economically and efficiently and inexpensively without complicated processes.

Γ-aminobutyric acid, abbreviated as GABA (GABA), is a nonprotein amino acid and is contained in grains such as human brain, vegetables, fruits, rice, and brown rice.

GABA is known to act as a neurotransmitter in higher animals, invertebrates and insects, including humans, and to act as a protective mechanism in plants.

In addition, as the effects of GABA on stress suppression, concentration enhancement, and blood pressure strengthening are known in the human body, it has been released as various food and animal and animal additives including health functional foods, and its range of application is gradually expanding.

In addition, it is known that there is an effect of suppressing disease occurrence by stress relief, improving productivity by promoting secretion of growth hormone, and improving the quality of livestock products including beef.

Specific examples of the effects of GABA in animals are to relieve stress during oral administration to animals such as pigs and chickens in groups to improve weight gain, and to reduce the high-temperature stress that occurs during summer to reduce egg quality. It is known to reduce the mortality by helping to alleviate transport stress through movement, and recently tried to produce branded livestock by feeding natural products such as mulberry leaves with high GABA content as livestock feed.

Methods for producing GABA in large quantities include synthetic methods, extraction methods, fermentation methods and enzyme methods. However, due to the specificity of the chemical process, the synthetic method is not suitable for animal feed, and the extraction method generates too much waste water and waste during the extraction process, and the manufacturing cost is too high. The fermentation method is too efficient to convert to GABA. It is low and in fact difficult to apply to mass production as an animal feed additive.

On the other hand, the enzyme method is a production method with a high possibility of being used as an animal feed additive, but as a factor to increase the production cost, the production process involves desalination, decolorization, filtration, concentration, crystallization process, and as a result, GABA Has been used only in high value-added fields such as raw materials for human health functional foods or medical fields, and it has been practically difficult to use as a feed additive for livestock.

In more detail, the production of GABA by the enzymatic method, -COOH bound to the α-carbon of L- glutamic acid is decarbonated by the catalysis of glutamic acid decarboxylase to γ-aminobutyric acid. Wherein the decarboxylase can be derived from a variety of microorganisms.

For example, glutamic acid dicarboxylase enzymes include E. coli, Lactobacillus plantarum, Lactobacillus brevis, Lactococcus lactis, Lactobacillus pentosus, Lactobacillus sakei, Listeria monocytogenes, Saccharornyces cerevisiae, Bacteroides fragilter bacterium, tuber Mycobacco nigra Can be obtained by separation and purification.

The higher the specific activity of glutamic acid dicarboxylase, the more preferable it is, and it is preferable that it is 500 u / g-cell or more.

The pH range in which the enzyme acts is about 3.5-6.0, which acts relatively broadly, but is most preferably within the range of 4.5-5.5. However, if the pH is below 3.5 or above 6.0, the enzyme reaction hardly occurs.

As a related art for GABA, Korean Patent No. 10-0857215 (registered on September 1, 2008), 'Method for producing high purity gamma aminobutyl acid using enzyme', and Korean Patent Publication No. 10-2009-0029723 (2009) 03. 23. published method of 'γ-aminobutyric acid or γ-aminobutyric acid-containing composition manufacturing method', and Republic of Korea Patent Publication No. 10-2012-0021399 (2012.03.09. Composition for addition, and a method for producing the same.

It is an object of the present invention to address the problems associated with the high manufacturing costs involved in applying GABA to feed additives.

An object of the present invention is to inexpensively produce GABA-containing feed additives by eliminating expensive processes such as desalting, decoloring, filtration, concentration and crystallization in producing γ-aminobutyric acid from L-glutamic acid by enzymatic reaction. I want to be able to.

According to the present invention, there is provided a method for producing a GABA-containing feed additive.

In a first embodiment of the GABA-containing feed additive manufacturing method according to the present invention, (A) mixing the adsorbent excipient 20-70% (v / w) of purified water compared to the adsorbent excipient to prepare a primary mixture (adsorbent Mixing of excipients and purified water); (B) mixing the L-glutamic acid with the primary mixture in a ratio of the volume of purified water to the weight of L-glutamic acid in a ratio of 100: 1 to 6: 1 to prepare a secondary mixture; (C) converting the L-glutamic acid to gammaaminobutyric acid (GABA) by mixing glutamic acid dicarboxylase and pyridoxal phosphate in the secondary mixture to cause an enzymatic reaction while maintaining the pH at 4.5-5.5. (Enzyme reaction step); And (D) drying the adsorbent excipient and the mixture of the enzyme reactants (drying step); It includes.

Preferably, the adsorbent excipient is at least one selected from powders of grain raw material, silica, zeolite, bentonite and vermiculite, and these powders may be used alone or in combination.

Preferably, the method for producing a GABA-containing feed additive according to the present invention further comprises the step of molding the dried mixture into pellets.

The adsorbent excipient is a component that adsorbs, holds, and gives shape to GABA. Preferably, the grain material may be used in the form of a powder having a particle size of approximately 1-3 mm, and an inorganic adsorbent excipient such as silica may be used in the form of a powder having a particle size of about 200 mesh.

As the grain raw material as the adsorbent excipient, powders of various feed grains and grain by-products such as degreasing rice bran, soybean meal, corn and the like can be used. When using a grain material having a high protein content or a high buffering capacity, the pH is adjusted to the above-mentioned range by mixing an acid as a pH adjuster as described below.

The purified water to be mixed as the reactive mass of the enzyme reaction is mixed at 20-70% (v / w), preferably 60% (v / w) relative to the adsorbent excipient.

When the content of the purified water is lower than the range in the mixing step (A) of the adsorbent excipient and purified water, the enzyme (glutamic acid dicarboxylase) and the coenzyme (pyridoxalphosphate) are absorbed between the particles of the adsorbent excipient and are insoluble. The contact with the L- glutamic acid is difficult, if higher than the above range has the disadvantage that the time and cost required for drying after the reaction increases.

The content of L-glutamic acid mixed in the primary mixture in L-glutamic acid mixing step (B) is preferably a ratio of the volume of purified water to the weight of L-glutamic acid in a ratio of 100: 1 to 6: 1, More preferably, the ratio is 20: 1 to 10: 1.

When the content of the L- glutamic acid is less than the ratio, a feed additive having a too low GABA content may be produced, and thus the economic value may be lowered. May be undesirably high.

In order for the enzymatic reaction of L-glutamic acid to proceed continuously in the enzyme reaction step (C), the pH should be maintained at 4.5-5.5, but if the pH is in the above range, it is not necessary, but if the pH is out of the above range, Acid may be adjusted as the pH adjusting agent to adjust the pH in the above range.

Although both inorganic and organic acids may be used as the pH adjusting agent, it is preferable to use organic acids in that the feed additive according to the present invention is intended to feed livestock.

Examples of the organic acid (pH regulator) include citric acid, tartaric acid, malic acid, lactic acid, acetic acid, butyric acid, propionic acid, fumaric acid, formic acid, itaconic acid, L-aspartic acid, and the like, and hydrochloric acid, sulfuric acid, phosphoric acid, and the like. have.

Preferably, the temperature of the enzymatic reaction is preferably maintained at 30-37 ° C, the enzymatic reaction is not lowered or extremely occurs outside the temperature range.

In order to maintain the temperature of the enzymatic reaction, the temperature may be maintained by heating from the primary mixture or from the secondary mixture.

In the drying step (D) may be applied hot air or vacuum drying method, and may be dried at a temperature of 20-200 ℃, preferably 40-90 ℃. If the drying temperature is lower than 40 ℃ takes too long drying time and higher than 90 ℃ has a disadvantage in that the sensory preference of the product is lowered because the color is changed due to degeneration of the carbohydrate included in the grain material.

On the other hand, the conventional enzymatic method is a method of preparing by stirring the reaction of purified water and L- glutamic acid and the reaction of the enzyme into a closed reactor in general, in order to remove a large amount of carbon dioxide bubbles generated in the enzymatic reaction The addition of surfactant (defoamer) was inevitable. At this time, if the bubble is not removed immediately, the reaction solution with the carbon dioxide gas was easily leaked out to the outside.

However, in the method of the first embodiment according to the present invention, since the enzymatic reaction proceeds in a mixed state with the adsorbent excipient, the loss of the reaction solution due to the generation of bubbles does not occur substantially, and thus it is not necessary to apply the antifoaming agent. Thus, the use of expensive surfactants is not necessary.

In addition, if the drying in the drying step (D) in a manner of continuously injecting the wind with a constant temperature with stirring, a space is formed between the particles of the mixture can be easily dried, thereby reducing the drying cost.

The method of the first embodiment according to the present invention is desalination, which is a complicated and costly process that was entailed in the prior art by enzymatic reaction proceeding in the presence of the adsorbent and the resulting GABA adsorbed directly to the adsorbent, Decolorization, filtration, concentration and crystallization processes can be eliminated, which makes it possible to produce GABA-containing feed additives at a significantly lower cost than conventional methods.

According to a second embodiment of the GABA-containing feed additive manufacturing method according to the present invention, (A1) preparing a mixed solution by mixing 1-40% (w / v) L-glutamic acid with purified water compared to the purified water (purified water and Mixing L-glutamic acid); (B1) converting the L-glutamic acid to gamma aminobutyric acid (GABA) by maintaining the pH at 4.5-5.5, causing an enzyme reaction by mixing glutamic acid dicarboxylase and pyridoxal phosphate in the mixed solution ( Enzyme reaction step); (C1) mixing 20-100% (w / w) of the enzyme reaction solution with the adsorbent excipient (adsorbent excipient mixing step); And (D1) drying the adsorbent excipient and the mixture of the enzyme reaction solution (drying step); It includes.

While the production method of the first embodiment according to the present invention is a method of carrying out an enzymatic reaction in the presence of the adsorbent excipient, the production method of the second embodiment according to the present invention uses the adsorbent excipient in the enzyme reaction solution in which the enzymatic reaction is completed. It is a way of mixing.

In a second embodiment, the mixing of purified water and L-glutamic acid in the mixing step (A1) of purified water and L-glutamic acid is preferably carried out in a liquid phase reactor. The total amount of the related components such as the purified water is preferably not more than 60-70% of the liquid phase reactor capacity (volume). If the input amount is larger than the above range, it is difficult to remove the carbonic acid bubbles generated in the enzymatic reaction step, which may cause the reaction solution to flow out.

The mixed content of L-glutamic acid is 1-40% (w / v) compared to purified water, preferably 30-35% (w / v).

When the content of the L- glutamic acid is less than the above range, a feed additive having a low GABA content may be prepared, and thus the economic value may be lowered. When the content of the L-glutamic acid exceeds the above range, the content of L-glutamic acid remaining without being converted to GABA is exceeded. This may be undesirably high.

In a second embodiment, the enzyme reaction step (B1) is a carbonic acid bubble is generated by the enzymatic reaction by mixing the glutamic acid dicarboxylase and the pyridoxal phosphate in a mixed solution of the purified water and the L- glutamic acid, such carbonic acid The remainder is substantially the same as the enzyme reaction step (B) of the first embodiment, except that an antifoaming agent can be preferably administered to remove the bubbles.

The adsorbent excipient mixing step (C1) is carried out in a stirrer, the mixing of the enzyme reaction solution and the adsorbent excipient obtained in the enzyme reaction step (B1), the enzyme reaction solution (GABA containing solution) compared to the adsorbent excipient 20 It can be mixed at -100% (w / w), preferably at 40-60% (w / w). If the mixing ratio of the adsorbent excipient and the enzyme reaction solution is lower than the above range, the GABA content is too low, and the feed additive is too low, and if it is too high, the drying efficiency in the drying step (D1) drops drastically.

The drying step (D1) of the second embodiment can be carried out by hot air drying or vacuum drying as in the first embodiment.

The method of the second embodiment is a method in which the adsorbent excipients are mixed so that GABA is adsorbed to the adsorbent excipients after the enzymatic reaction is completed. However, without the process of desalting, bleaching, filtration, concentration, crystallization, GABA-containing feed additives according to the present invention can be prepared.

The method for preparing a GABA-containing feed additive according to the present invention includes a process of converting L-glutamic acid into GABA by adsorption of glutamic acid dicarboxylase and adsorbing GABA to an adsorbent excipient into one series. As a method to be carried out in the process, first compared to the conventional method of producing GABA from L-glutamic acid as a finished product using glutamic acid dicarboxylase as a catalyst, and then processing GABA again as a feed additive in a separate process. By eliminating complex and costly processes such as desalting, bleaching, filtration, concentration and crystallization associated with conventional methods, the associated manufacturing equipment can be significantly simplified, resulting in GABA-containing feed compared to conventional methods. It is possible to supply additives at a significantly lower cost, which is also useful for many useful effects on plants and animals. It is possible to realistically make use of GABA as a feed that could not be obtained due to its high price, and consequently, through the wide application of GABA to feed, it is necessary to raise a large number of livestock in a small space with feed for high productivity. It is expected to be useful for mitigating fatal losses caused by disease infections in poor breeding environments.

In addition, the manufacturing method of the GABA-containing feed additive according to the present invention, since the desalination, decolorization, filtration, concentration, crystallization process is not applied, there is an effect that can reduce the generation of waste water to prevent environmental pollution and reduce the related costs. .

Figure 1 is a photographic view of the GABA-containing feed additive according to the present invention (A in Figure 1 powder form, Figure 1B is a pellet form),
Figure 2 is a photographic view of the enzyme reaction solution in Example 2.

Hereinafter, a method for preparing a gamma aminobutyric acid-containing feed additive according to the present invention through specific examples will be described in more detail. The following embodiments are merely illustrative of the present invention and are not intended to limit the scope of the anti-invention.

Example 1 . GABA  Preparation of Feed Additives Containing

Example 1 was manufactured using a stirrer equipped with a hot air supply device for supplying hot air at a desired temperature.

First, 100 kg of degreasing rice bran, which is an example of a grain material as an adsorbent excipient, and 60 L of purified water, which is a reactive substance, are added to the stirrer, and 10 kg of L-glutamic acid is uniformly mixed with the introduced purified water and the adsorbent excipient (primary mixture). It injected | threw-in and fully stirred (mixed) (secondary mixture). At this time, it was heated to maintain the temperature of the secondary mixture at 37 ℃.

In the secondary mixture, enzymatic reaction was initiated by adding and stirring pyridoxalphosphate 0.02 mM as a coenzyme and 500 g of glutamic acid dicarboxylase as a decarboxylase, respectively.

After the enzymatic reaction was started, stirring was continued for about 15 hours to complete the enzymatic reaction, and the mixture of the adsorbent excipient and the enzyme reactant was dried by hot air drying. At this time, the drying conditions were the temperature 70 ℃, stirring speed 30rpm, drying time 32 hours. The feed additive in the form of a powder (see FIG. 1) according to the present invention after the completion of drying was measured at 8.3% by weight of water and 9.04% by weight of GABA.

Example 2 . GABA  Preparation of Feed Additives Containing

A 100 L capacity liquid phase reactor was used as the reactor for the enzymatic reaction, and 60 L of purified water and 18 kg of L-glutamic acid as reactor materials were added to the liquid phase reactor and stirred (mixed), and the temperature of the mixed solution was maintained at 37 ° C.

The enzymatic reaction was initiated by adding and stirring 0.02 mM of pyridoxalphosphate as a coenzyme and 900 g of glutamic acid dicarboxylase as a decarboxylase to the mixed solution of the liquid phase reactor.

Tween 80 (Junsei, Japan), a surfactant, was diluted with water at a rate of 10% (w / v) and intermittently added to remove carbonic acid bubbles generated in the enzymatic reaction.

78 kg of the enzymatic reaction solution (see Fig. 2) obtained as a result of the enzymatic reaction was transferred to a stirrer as in Example 1 with a hot air supply device, and then 100 kg of degreasing rice, an example of a grain material as an adsorbent excipient, was added and stirred ( Mixed).

The mixture of the enzyme reaction solution and the adsorbent excipient was dried by hot air, and the drying conditions were a temperature of 70 ℃, stirring speed 30rpm, drying time 42 hours.

The feed additive in powder form according to the present invention (see FIG. 1), which was dried and finished, was measured to have a water content of 9.2% by weight and a GABA content of 25.12% by weight.

Example 3 . GABA  Preparation of Feed Additives Containing

Into a 100L capacity liquid phase reactor as in Example 2, 60 kg of purified water and 18 kg of L-glutamic acid were added and stirred (mixed) as a reactor, and the temperature of the mixed solution was maintained at 37 ° C.

The enzymatic reaction was initiated by adding and stirring 0.02 mM of pyridoxalphosphate as a coenzyme and 900 g of glutamic acid dicarboxylase as a decarboxylase to the mixed solution of the liquid phase reactor.

In addition, by diluting the surfactant tween 80 with water 10% (w / v) intermittently to remove the carbonic acid bubbles caused by the enzyme reaction.

The enzyme reaction solution was transferred to the same stirrer as in Example 1 with a hot air supply device, and then dried by hot air drying while adding and stirring a powder of zeolite (200 mesh) as an adsorbent excipient.

At this time, the enzyme reaction solution and zeolite (adsorbent excipient) were mixed at a ratio of 1: 1 by weight. Drying conditions were the drying temperature of 70 degreeC, the stirring speed of 30 rpm, and the drying time of 38 hours.

The feed additive in powder form according to the present invention (see FIG. 1) completed after drying was determined to have a water content of 8.4% by weight and a GABA content of 22.15% by weight.

Example 4. Preparation of high concentration GABA-containing feed additive (pellet type)

Prepare a feed additive in the same manner as in Example 2, but transfer the enzyme reaction solution 50kg of enzyme reaction is completed to the same stirrer as in Example 1 equipped with a hot air supply device, and then corn as an example of a grain material as an adsorbent excipient 100 kg of powder was added and mixed (stirred). Moisture content at this time was measured as 14.8% by weight.

A mixture of the adsorbent excipient (corn powder) and the enzyme reaction solution was dried with hot air to prepare a feed additive in powder form according to the present invention. At this time, the drying conditions, the temperature was 70 ℃, stirring speed 30rpm, drying time was 28 hours.

The dried feed additive of the present invention was formed into pellets having a diameter of 6 mm by using a circular pellet molding apparatus to complete the pellet-type feed additive according to the present invention (see FIG. 1), and according to the final invention after pellet molding Feed additives were measured as 12.04% by weight of water and 17% by weight of GABA.

When the moisture content of the grain material as an adsorbent excipient is about 14-20% by weight, the pellet feed additive may be prepared without adding additional moisture or using a binder for pellet molding.

Claims (4)

Preparing a primary mixture by mixing 20-70% (v / w) of purified water with respect to the adsorbent excipient;
Preparing a secondary mixture by mixing the L-glutamic acid with the primary mixture in a ratio of the volume of purified water to the weight of L-glutamic acid in a ratio of 100: 1 to 6: 1;
converting the L-glutamic acid to gammaaminobutyric acid (GABA) by enzymatic reaction by mixing glutamic acid dicarboxylase and pyridoxalphosphate in the secondary mixture while maintaining the pH at 4.5-5.5; And
Drying the adsorbent excipient and the enzyme reactant;
Characterized in that it comprises a GABA-containing feed additive. Preparing a mixed solution by mixing 1-40% (w / v) L-glutamic acid with purified water compared to the purified water;
maintaining the pH at 4.5-5.5, converting the L-glutamic acid to gamma aminobutyric acid (GABA) by mixing glutamic acid dicarboxylase and pyridoxal phosphate in the mixed solution to cause an enzymatic reaction;
Mixing 20-100% (w / w) of the enzyme reaction solution with the adsorbent excipient; And
Drying the adsorbent excipient and the enzyme reaction solution;
Characterized in that it comprises a GABA-containing feed additive. The method according to claim 1 or 2,
The adsorbent excipient is characterized in that at least one selected from powders of grain raw material, silica, zeolite, bentonite and vermiculite, GABA-containing feed additive manufacturing method. The method according to claim 1 or 2,
Method for producing a GABA-containing feed additive, characterized in that it further comprises the step of molding the dried mixture into a pellet shape.

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