KR100388109B1 - Method for producing taurine enriched milk - Google Patents

Abstract

The present invention relates to a method for producing milk of an animal having a high taurine content, and more particularly, to prepare a precursor of taurine synthesis in the form of pellets or a matrix stabilized against potential degradation of ruminants, and to feed the substrate, The present invention relates to a method of producing milk enriched with natural taurine by in vivo metabolism by stimulating biosynthesis of taurine. The method of the present invention coats tau with methionine or methionine hydroxy analog, which is a precursor of taurine synthesis, and protects it from ruminant pH and ruminant enzymatic activity by coating with additives such as inerts and fillers. When directly supplied, it solved the problem that taurine was decomposed in the rumen or interfered with digestion metabolism after ruminant metabolism or ruminant, and the taurine fortified milk of the present invention contained taurine to a higher degree than ordinary milk, so that functional milk for infants or health It can be effectively used for the production of food.

Description

Method for producing taurine enriched milk {Method for producing taurine enriched milk}

The present invention relates to a method for producing milk of an animal having a high taurine content, and more particularly, to prepare a precursor of taurine synthesis in the form of pellets or a matrix stabilized against potential degradation of ruminants, and to feed the substrate, The present invention relates to a method of producing milk enriched with natural taurine by in vivo metabolism by stimulating biosynthesis of taurine.

Taurine (taurine; 2-amino-ethanesulfonic aicd) is one of the sulfuric acid amino acids biosynthesized in the body of mammals and exists freely in the intracellular fluid, especially skeletal muscle of the brain, eye, muscle, liver, etc. It is found in high concentrations in the free amino acid family of the nervous system, and has been used in food and beverages for a long time as a healthy material with various functions such as enhancing liver function, reducing blood cholesterol level and controlling blood pressure through detoxification and antioxidant activity. Taurine contains bile acids in the carnivorous body and is excreted through bile in the form of taurocholic acid. Taurine released into bile promotes the absorption of fat or fat-soluble vitamins in the intestine by interfacial action. It has the effect of reducing jaundice during acute hepatitis and lowering the level of triglyceride and cholesterol in serum of hyperlipidemic patients. In addition, it develops brain, photoreceptor activity, reproductive and normal growth development, antioxidant activity, It has been reported to have various functions such as neurotransmission (Jacobsen et al. , Physiol. Rev. 48: 424-511, 1968).

Due to this physiological activity, taurine is used as an additive to various health foods and medicines such as baby food, gum, beverages, and nutrient tonic drinks, but it is not frequently used as a drug, and therapeutic use is reported only for the treatment of heart disease. .

In mammalian tissues, taurine is biosynthesized from the sulfur-containing amino acids cysteine and methionine.In the human body, the activity of cysteine dioxygenase and cysteine decarboxylase is involved in the biosynthesis of taurine. This very low biosynthesis of taurine is rarely achieved and must be supplied from outside (Vinton et al ., Pediatr. Res . 21: 399-403, 1987).

On the other hand, taurine maintains a high concentration at almost constant levels in mammalian tissues, but the taurine content in milk shows a great difference depending on the type of animal. Taurine concentrations in the milk of various types of mammals were measured (Rassin et al ., Early Hum. Dev. 2 (1): 1-13, 1978; Sarwar et al., British) . Journal of Nutrition 79: 129-131, 1998), zebras and cats contain 595μmol / 100ml and 287μmol / 100ml taurine, respectively, the highest concentration of free amino acids in milk and 34μmol in breast milk. Contains 100 ml of taurine, the next highest concentration after glutamate. On the other hand, rats, rabbits and sheep are contained in 14-15μmol / 100ml of taurine, and cows are known to have 1μmol / 100ml of taurine, which is the lowest in mammalian milk. In particular, in the case of breast milk, taurine is the main amino acid, which accounts for about 13% of the total free amino acid, while milk, which is the main ingredient of the formula, has a relatively low taurine content.

Because of the low taurine content in milk, there are no clinical reports of taurine deficiency in adults who normally eat food, but premature infants and infants who do not have taurine supplemented formula are more likely to receive plasma than infants who consume breast milk. It has been reported that taurine levels have been reduced (Rassin et al ., Pediatrics 71: 179-186, 1983; Rigo et al. , Biol. Neonates 32: 73-76, 1997). Infants also have a low reuptake capacity of taurine due to immaturity of the renal tubules (Zelikovic et al. , J. Pediatr. 116: 301-306, 1990), while taurine demand in the body increases due to the rapid growth of the body. As a result, the need for taurine is emerging for healthy growth and development. In particular, infants have a weaker ability to digest and absorb fat because of their weaker synthesis, secretion and reuse of bile acids in the body than adults. Therefore, there is insufficient emulsification of fats necessary for rapid breakdown of ingested fats by pancreatic lipase. In addition, since bile acid-stimulating lipases are contained in breast milk, it is possible to efficiently break down fats jointly with pancreatic lipases even in the absence of bile acids, but infant formulas do not contain bile acid-stimulating lipases. Therefore, in order to overcome the problem of formula milk, in the United States, taurine, which is effective in promoting the absorption of fat or fat-soluble vitamins by surfactant activity, is added to formula milk at the level of breast milk (Picone et al ., Nutr. Today July / Aug: 16-20, 1987), taurine is additionally fortified in the formula of infant formula currently available in Korea.

The production method of taurine which is used for various purposes includes natural taurine extraction method and chemical synthesis method. The former method of extracting natural taurine is the method of extracting taurine from molluscs such as squid, octopus and shellfish in which a large amount of taurine exists. Products produced by natural taurine extraction method can be used for food and medical purposes, but all Very small amount of taurine, such as 1% compared to taurine products, mainly using chemical synthesis method.

There are various methods of the latter chemical synthesis method, which may contain other impurities in the production of synthetic taurine, and the impurities cannot be completely removed in the process of refining them, and thus the remaining impurities The use is limited and in particular, inexpensive synthetic taurine cannot be used for food.

Therefore, the necessity of natural taurine, which can be used for a variety of purposes such as food and medicine, is increasing, and a method of producing taurine-containing products by recovering the waste liquid of squid stripping liquid and cooked liquid generated in the squid processing process or (Korean Patent Publication No. 00 -58466) Research has been made on the method of producing dry eggs by pulverizing the shell part discarded except for the decorative part of the crab, and then feeding it to the feed (Korean Patent Registration No. 96- 16856).

However, there is a problem in the method of producing a product such as food containing a large amount of taurine by directly adding taurine to the product or by directly supplying a feed containing abundant taurine as in the above method. First, the problem of adding taurine directly to the product is that the amount of taurine supplied by the natural taurine extraction method is very small, as described above, so that productivity and economic efficiency may be reduced.

In addition, the problem of the method of directly supplying a feed rich in taurine is that when applied to a ruminant animal, taurine is decomposed by microorganisms present in the rumen to produce a product containing the desired taurine. It is difficult. In general, the nutrient digestion process of ruminant ruminants consists of the first stage fermentation process by microorganisms in the rumen and the two stage digestion process in the lower digestive organs. It is composed of the sum of the nutrients of the microorganisms and the nutrients that have passed into the small intestine without being broken down in the first place. On the other hand, the rumen microorganisms have strong protease and deminase activity to rapidly break down soluble proteins and amino acids in the liquid rumen, so that the precursor amino acids of taurine or taurine synthesis in the feed There is a risk of decomposition within the potential. Therefore, much research has been conducted on methods of reducing the degree of degradation of proteins and amino acids in the rumen: selective manipulation of heat treatment, chemical treatment, use of amino acid analogs, encapsulation, esophageal groove closure, ruminant metabolic pathway balance Methods are known (Chalupa et al ., J. Dairy Science 58 (8): 1198-1218, Chen et al ., Analytica Chimica Acta 296: 249-253, 1994).

Another problem with the method of adding taurine directly to the feed is that if excess taurine is present in vivo, it may interfere with ruminant metabolism and post- ruminant digestion absorption.

In order to solve the problems of the conventional taurine supply and to manufacture milk with increased taurine content, the present inventors induce the taurine synthesis in vivo by preparing and feeding a precursor of taurine synthesis in a form having resistance to rumen breakdown. It was confirmed that taurine could produce fortified milk. Precursors of taurine synthesis are sulfur-containing amino acids such as cysteine and methionine, all of which are essential for animal protein metabolism and in particular, are evaluated as the first limiting amino acid for milk protein synthesis. The amount of cysteine and taurine is very low in ruminant tissue, blood, urine and milk compared to the unit animal. There are genetic and physiological causes, but ruminants are all herbivores and the plant proteins they consume are taurine and taurine. Due to the lack of a source of supplementation, supplementation of methionine against rumen breakdown that can be absorbed in the small intestine rather than breakdown in the rumen can be expected to increase the taurine content with changes in milk protein composition due to substrate stimulation. . In other words, protection from rumen bypass and rumen degradation is important for the small intestine absorption of the amino acids involved in taurine synthesis and secretion through milk, and chemical treatment and encapsulation methods are expected to be effective. It became.

Although many studies have been conducted on pellets or methods for producing pellets containing active ingredients such as amino acids protected against ruminant degradation of ruminants, or animal feed containing the pellets, these are all ruminants through the supply of the active ingredients. Focusing on the improvement of breeding methods (Korean Patent Publication No. 95-27379, Korean Patent Registration No. 143766, US Patent No. 5,279,832, US Patent No. 4,181,708, US Patent No. 5,776,483) There has been no report on milk production with increased taurine content.

Therefore, in the present invention, by supplying a pellet or matrix containing amino acids protected against dislocation degradation of ruminants by using a metabolic pathway in vivo to feed the taurine-enriched milk through the induction of taurine synthesis by substrate stimulation To provide a way to produce.

An object of the present invention is to provide a production method for increasing the content of taurine deficient in milk such as milk. Specifically, an object of the present invention is to overcome the fear of degradation in the rumen, which is a problem of the existing natural taurine direct supply and the interference of ruminant metabolism or digestion metabolism after rumination, and supplies taurine precursors resistant to rumen degradation It is thereby to provide a method of producing taurine fortified milk.

It is also an object of the present invention to provide milk having a high taurine content by the above method. The natural taurine fortified milk of the present invention is expected to be effective as a functional milk that is beneficial to infants with high taurine demand due to enhanced taurine lacking in milk compared to breast milk.

1 shows the biosynthetic pathway of taurine produced from precursors such as methionine,

Figure 2 is a measure of the degree of protection in the simulated buffer to investigate the degree of protection against pH in the rumen after supplementation of protective amino acids, the white bar is the effluent rate of RP (rumen protected) DL-methionine, the black bar is RP Shows the outflow rate of MHA (methionine hydroxy analog),

Figure 3 is a measure of the degree of protection in imitation buffer containing rumen contents to investigate the degree of protection against digestion of ruminant microorganisms after the protection amino acid supplement, the white bar is the effluent rate of rumen protected (DL) methionine , Black bars indicate the outflow rate of RP MHA (methionine hydroxy analog),

4 is a chromatogram of tau DL content of crude milk samples of the specification cow after feeding RP DL-methionine (a) and tauine content (b) after feeding RP MHA with an amino acid-only analyzer,

Figure 5 is the result of measuring the change in the concentration of taurine in milk after feeding RP DL-methionine (a) and the result of measuring the change in the concentration of taurine in milk after feeding RP MHA (b).

In order to achieve the above object, the present invention provides a precursor of taurine synthesis that is resistant to ruminant gastric degradation and feeds it to the animal along with the rich feed to induce taurine biosynthesis through substrate stimulation to obtain milk of high taurine animal content. Provide a way to produce.

The present invention also provides a milk with a high content of taurine by the above method.

In the present invention, "mature milk" refers to a normal milk having a milk composition and a flow rate of about 300 days, which are secreted after colostrum in which cows secrete 4-7 days after delivery, and "protection." Amino acid "refers to a coating of taurine biosynthetic precursor amino acids or derivatives thereof with additives such as insoluble substances and fillers so as to be protected from degradation due to the enzymatic action of ruminants. In a preferred embodiment of the present invention, "RP DL-Met" (rumen protected DL-methionine) or "RP MHA" (rumen protected methionine hydroxy analog) was used as an example of a specific protective amino acid. In addition, "ruminant degradation" means the degradation of soluble proteins and amino acids due to the pH and ruminant proteolytic and deamino activity of the rumen.

Hereinafter, the present invention will be described in detail.

The method of producing milk with a high taurine content of the present invention,

1) preparing a precursor of taurine biosynthesis in the form of a protective amino acid that is resistant to rumen degradation;

2) raising livestock by limiting the feed containing the protective amino acid; And

3) harvesting the crude oil from the livestock.

In the above, a precursor of taurine biosynthesis is preferably a methionine or methionine analogue, and the methionine analogue is N-acetyl-DL-methionine,

Benzoyl-L-methionine, N-phthalyl-DL-methionine (N-phthalyl

N-propionyl-DL-methionine, N-carbobenzoxy-DL-methionine, Nt-butyloxycarbonyl L-methionine-dicyclohexyl ammonium salt (Nt-butyloxycarbonyl-L-methionine-dicylohexyl ammonium salt) Nt-butyloxycarbonyl-L-methionine-p-nitrophenyl-DL-methionine (Nt-butyloxycarbonyl-L-methionine- p-nitrophenyl-DL-methionine)) (W. Chalupa, J. Dairy Sci . 58 (8): 1198-1218, 1974). As a preferred methionine analog, 2-hydroxy-4-methylthio butanoic acid (C ₅ H ₁₀ O ₃ S) may be used, wherein the amine group of the methionine is a hydroxyl group. It has a substituted chemical structure and is metabolized to methanethiol and dimethylsulfide, which is known to be similar in metabolism of methionine and ruminant microorganisms. MHA has the advantage of low solubility in ruminant fluidized bed, somewhat resistant to ruminant microorganisms compared to DL-methionine, low cost and water soluble, and easy application in manufacturing process. However, the pH of about 1.0 has a strong acidity, so the neutralization by calcium hydroxide is required, so a large amount of calcium hydroxide is used compared to the production of RP DL-methionine, which has a disadvantage of decreasing the content of the central substance.

The source of amino acids absorbed by the small intestine of ruminants is by protein and synthetic endogenous secretion by the rumen bypasses, and in the case of endogenous secretion directly affects amino acids. It can't be crazy, but the other two cases can be artificially adjusted.

In the method of producing milk with a high content of taurine, a method of making amino acid, which is a taurine precursor, has resistance to rumen degradation, may include spray drying and fluidized bed drying by incorporating additives such as insoluble materials and fillers into methionine. Fludized bed drying, extrusion, pelletizing and the like can be used.

Methionine, which is supplied as a precursor of taurine synthesis in milk, of the present invention, is digested and absorbed in the small intestine by passing through the rumen, and then reaches the mammary tissue in the rumen of pH 5.3 for 10 to 30 hours in order to reach the mammary tissue. It must be stable to pH and pH while eluting within 4 hours at pH 2.9 to meet the conditions for absorption in the small intestine.

In a preferred embodiment of the present invention, a pelletization process was used as a manufacturing method to have a protective action against rumen degradation. Cured beef tallow (hydrogenated beef tallow) can be used in the pelletization process, and the hardened beef tallow is inexpensive as a main ingredient that makes particles insoluble, and has a property of solidifying at 60 ° C. or less. Can be achieved. In addition, the pellet may include calcium hydroxide (Ca (OH) ₂ ), which increases the pH of the core material and provides firmness when combined with bee tallow, which greatly affects the solubility in the rumen. have. In addition, the pellet may contain an emulsifier and an antifoaming agent. In a preferred embodiment of the present invention, a polyglycerol fatty acid ester emulsifier is used as the emulsifier, and the emulsifier has an effect of emulsifying a tallow and gelatin solution. In addition, the antifoaming agent has an antifoaming and emulsifying effect when preparing a gelatin solution. In addition, the pellet may include vitamin E, which is a gelatin and an antioxidant that acts as a binder of tallow and methionine.

As an example of a preferred protective amino acid for producing taurine fortified milk, the protective methionine is formulated with about 60 to 70% by weight of methionine, about 20 to 25% by weight of tallow, about 5 to 10% by weight of starch, and 2 to 4% by weight of gelatin. It is preferable to produce a sphere having a diameter of about 3 to 6 mm, a specific gravity of about 1.0 to 1.05, which is a particle size stable to physical and mechanical impacts. That is, by having the composition and form as described above it can be made easy to transport and preservation and application with other feed, it is possible to minimize the destruction of particles by chewing, increase the amount of taurine precursor absorbed in the small intestine. The composition may additionally be prepared by adding calcium hydroxide, surfactants, antioxidants and antifoaming agents, in addition to adding propylene glycol for uniform mixing of methionine and matrix material, or sugar palmitic acid to adjust the solidification rate of tallow A larger effect can be obtained by adding an ester or the like.

The method for producing the pelletized protected amino acid is preferably emulsifying hydrogen cured tallow and gelatin solution at a temperature above the melting point, mixing well with methionine, solidifying and mixing for about 1 hour to obtain uniform crystal grains, and adding starch. And then molded in the form of pellets and dried at 50 ~ 60 ℃ for more than 25 hours includes the process of making the moisture content less than 5%. If methionine hydroxy analogs are used instead of methionine, it is preferred to add a process of neutralizing with calcium hydroxide to pH 5.0 or higher first. Dextrin may be used instead of starch or gelatin.

In addition to the pelletizing method as described above, spray drying, extrusion molding, fluidized bed drying, etc. may be used. In the case of spray drying, the encapsulation method for rumen protection is the best method, but because methionine is insoluble, O / W emulsification is difficult and even if emulsification has a problem that the content of the central material is less than 20%. In addition, in the extrusion method, the melting point of the Uji used as an insoluble material may cause cracks in the particles in the heating step, which may cause fracture of the particles, and also have weaknesses in bonding with methionine during extrusion. On the other hand, in the case of fluidized bed drying, the particle size of methionine is different from the coating material, so that it is difficult to obtain uniform particles, and the processing time is long and the processing cost is high. Therefore, it can be seen that it is most preferable to use the above-mentioned pelletization method in order to promote the binding of the coating material such as fatty acid and the coating material such as filler and methionine which is the core material and to obtain uniform particles.

In raising livestock by feeding the feed containing the precursor of taurine synthesis, it is preferable to use a method of maintaining a constant intake by implementing a limited feeding method of feeding the additives with the rich feed only when milking. Preferably, the taurine precursor prepared to resist rumen degradation may be fed to 0.1 to 1.0% of the total daily feed amount of the feed.

Taurine fortified milk that can be produced by the method of the present invention is applicable to all of the milk of various mammals such as goat milk, goat oil, sheep milk, but when applied to the milk (milk) used for the preparation of infants with a large consumption of infants The effect is even greater.

In order to increase the content of taurine in milk, it is important that the protective amino acids prepared by the method have biologically easily absorbed properties while being resistant to ruminant microorganisms and chemical degradation of the rumen. Therefore, the degree of protection was investigated in the buffer solution containing rumen contents in order to know the degree of degradation and leakage of the enzyme produced by the ruminant microorganism of the matrix prepared by the above method. The degree of protection is the same as measuring the outflow of active ingredient in the simulated buffer solution minus the degree of outflow observed in vitro from 100 and the degree of outflow is expressed as the percentage ratio of active ingredient outflow during the test. As a result of the test, RP-Met was protected about 60-64% in the rumen while RP-MHA was more than 70%, and after investigating the efflux effect after the passage of the rumen, the fourth of RP-Met and RP-MHA was examined. The outflow rate in gastric and intestinal gastric fluid was about 90%. Thus, the protective amino acid supplement of the present invention can lead to an increase in induction of taurine synthesis in vivo.

In addition, the taurine content in milk was increased due to the increase in taurine synthesis in the body by the supplementation of the protective amino acids RP-Met and RP-MHA.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example 1 Preparation of DL-Methionine Protected Against Rumen Degradation

As taurine precursors were prepared protective amino acids that were effectively broken down in the small intestine without degradation in the rumen. In the preparation of protective amino acids, the coating material is a bio-applicable material that has no effect on metabolism and nutrition. In addition, fatty acid was formulated to resist the enzymatic action of rumen microorganisms, and it was spherical in particle size φ 3-6 mm and specific gravity 1.0 ~ 1.05 that is stable to physical and mechanical impact. It was prepared to facilitate the application of and to minimize particle destruction by chewing. The composition of the protective methionine matrix is shown in Table 1 below.

Constituents of the DL-Methionine Matrix Protected Against Rumen Gastric Degradation Ingredient % DL-Methionine 65.2000 Uji 24.2000 Starch 7.0000 gelatin 2.7000 Calcium hydroxide 0.4194 Twin 80 0.3600 Vitamin E 0.1170 Antifoam 0.0036 sum 100.0000

First, emulsify the hydrogenated beef and gelatin solution at a temperature above the melting point, mix well with methionine, solidify, and then mix with a mixer (KMX-3R Mixer, KOECO Industrial Co. Ltd.,) for at least 1 hour to obtain uniform crystal grains. Next, starch is added to form granular machine (rotary granulator 3HP, Youngjin machine Co. Ltd.) and dried at 50 ~ 60 ℃ for more than 24 hours to protect the moisture content of rumen of matrix embedded type with less than 5% moisture content. The prepared DL-methionine was used for the specification test, and the DL-methionine content of the prepared particles was 63% as determined by the iodine titration method of Example 3-1).

Example 2 Preparation of MHA Protected Against Rumen Degradation

The composition of the protective methionine hydroxyl analog matrix is Table 2 is as follows.

Ingredient % MHA 55.492 Uji 27.732 Calcium hydroxide 8.158 Twin 80 8.158 dextrin 0.460 sum 100.000

As a methionine hydroxy analog, Alimet Novus International, Inc., USA was purchased, neutralized with calcium hydroxide, the pH was adjusted to 5.0 or higher, and then mixed with Uji and Dextrin for at least 1 hour (KMX-3R). Mixer, KOECO Industrial Co. Ltd.,), followed by drying for more than 24 hours to bring the moisture content to less than 5%. The MHA content of the prepared RP MHA matrix was more than 57% as determined by iodine titration method, which is 12% water content of MHA seems to be the result of the moisture of the MHA itself removed through the drying process.

Example 3 Protection Confirmation Experiments on Ruminant Gastric Decomposition

3-1) Iodine (I ₂ ) DL-methionine titration by

Dissolve 0.2-0.6 g of the sample in distilled water to make 100 ml, add 50 ml to a revolving flask, add 50 ml of distilled water, ₂ g of potassium phosphate (KH ₂ PO ₄ ), and 2 g of KI to dissolve, and add 50 ml of 0.1NI ₂ solution. After leaving for 30 minutes, 2 to 3 drops of starch solution were added as an indicator, and titrated with 0.1 N sodium hydroxide (NaS ₂ O ₃ ㆍ 5H ₂ O). 1 ml of 0.11 I _{2 corresponds} to 7.461 mg methionine (C ₅ H ₁₁ O ₂ NS), which was used for protection against ruminant degradation and runoff assessment. In the case of MHA, the purity of MHA was adjusted to 88% in the same manner as described above.

3-2) Evaluation of Protective Effect and Runoff Activity against Rumen Protection

3-2-1) pH Stability

In order to measure the outflow rate in the first stomach, 5 g of the sample was put in 100 ml of the first stomach buffer solution, incubated with stirring at 37 ° C. for 16 hours, and the supernatant was taken to measure the methionine content by iodine titration.

In order to measure the outflow rate in the fourth phase, after measuring the outflow rate in the first phase, the remaining solid part was separated, and 5 g of the sample was added to 100 ml of buffer solution, followed by incubation with stirring at 37 ° C. for 2 hours, followed by iodine titration. In order to measure the outflow rate in the gastric juice, the remaining solid part was separated after the fourth outflow rate experiment, and 5 g of the sample was added to 100 ml of the small intestine gastric juice solution and reacted with stirring at 37 ° C. for 2 hours. The methionine content was measured by the following iodine titration method. As a result, the degree of leakage of RP DL-methionine and RP MHA in the fourth and small intestinal gastric fluid was about 90% (see FIG. 3).

3-2-2) Stability against ruminant microorganisms

In order to investigate the stability and outflow rate of the matrix against the enzymatic degradation of ruminant microorganisms, the contents of the rumen of Holsteine cow were extracted immediately after slaughter, and added to the simulated buffer solution by 12%, followed by culturing CO ₂ in the same manner as above. The protective effect and efflux of RP DL-methionine and RP MHA matrix were investigated. The degree of protection was expressed as the ratio of the amount of amino acid exempted from the amount of amino acid effluent to the total amount of the protected amino acid supplemented. As a result of the investigation of the degree of protection in the buffer solution containing the rumen contents, the RP DL- About 60-64% of the methionine was protected in the rumen, while more than 70% of the RP MHA was protected (see FIGS. 2 and 3). These results appear to be based on the amount of Uji used in the preparation of protective amino acids for the sake of robustness and solubility. The composition of the simulated buffer solution is shown in Table 3.

1st buffer solution 1ℓ 4th buffer solution 1ℓ Duodenum 1ℓ Na ₂ O ₄ 2.5 g 3.2 N KCl 50 ml NaHCO ₃ 9.8 g K ₂ HPO ₄ 6.7 g 0.2 NHCl 10 ml KCl 0.57 g 0.47 g Na ₂ HPO ₄ Na ₂ SO ₄ 7H ₂ 0 0.12 g 0.05 g of bovine bile powder 0.05 g of lipase (EC 3.1.1.3) pH 6.4 pH 2.0 pH 8.2

Example 4 Specification Test by Feeding Protective Amino Acids Against Rumen Degradation

4-1) Disclosure Livestock and Test Period

A six-week specification test was conducted with 10 or 12 holstein cows for the RP-Met or RP-MHA matrix feeding test. Each cow was divided into two groups, each with a control diet and 60 g / day of RP-Met matrix or RP-MHA matrix, with a limit of 15 kg / day of rich feed. Crude oil samples for each subject were collected before and after the supplementation.

4-2) Effect of Protective Amino Acid Supplementation on Milk Taurine Concentration

To analyze the taurine content of crude oil samples (MAMehaia, Milchwissenschaft , 47 (6), 1992, 351-353) reported by Mehaia et al. The sample was used as a taurine analysis method.

500 μl of a milk sample was taken in a 1.5 ml micro eppendorf tube, and 125 μl of a 20% sulfosalicylic acid solution was added thereto, followed by vortexing, and left at 4 ° C. for 60 minutes. The protein was removed by sedimentation by centrifugation at 14,000 × g for 10 minutes, the supernatant was taken, transferred to a clean tube, and filtered using a 0.2 μm filter (PVDF Aerodisc 13, Gelman Sciences) immediately before injection into the amino acid analyzer. Amino acid concentrations in milk samples were determined using an amino acid-only analyzer (Biochrom 20, Pharmacia LKB Biotech, Cambridge, England) based on an ion-exchange column (Moore et. Al., Academic Press, New York Vol 6: 819-831 1963). Measured. 40 μl of the pretreated sample was injected into a lithium high performance column (90 × 4.6 mm Pharmacia LKB Biotech) of an amino acid analyzer and a lithium citrate buffer in which the pH and ion concentration were gradually increased in the mobile phase. buffer) was used at a flow rate of 20 mL / hour (0.2M pH 2.80; 0.30M pH 3.00; 0.50M pH 3.15; 0.90M pH 3.50; and 1.65M pH 3.55).

Each amino acid separated through the column was reacted with ninhydrin to form a purple color, and then absorbance was measured at 440 nm and 570 nm, respectively. The resolution time of taurine was about 10.1-10.5 minutes (see Fig. 4). The amino acid concentration of each sample was obtained by injecting 20 μl of 0.5 mM standard amino acid solution (Sigma # A-6407 A-1585). The area of (peak) was calculated by comparing the area of the peak obtained from each sample. As shown in FIGS. 4a and 4b, the strong acidic phosphoserine first exited the column, followed by taurine, phosphoethanolamine, and urea, and took a total of 188 minutes to separate. It took a total of 237 minutes for the column to be washed, refurbished and the next sample injected. Taurine was separated into a single peak that was completely separated from the phosphoethanolamine and phosphoserine peaks, and showed an unknown peak before the taurine peak. The error coefficient of the repeated experiment was within 5%, and norleucine was added to the sample as an internal standard to compensate for the loss generated during the pretreatment and analysis of the sample.

In the RP-Met feeding test, the taurine concentration of the control group was 34.78 ± 8.91 μmol / l in the pre-feeding sample and 33.38 ± 6.32 μmol / l after 42 days, and 36.53 before the feeding in the RP DL-Met matrix feeding group. ± 2.83 μmol / l, 44.77 ± 5.61 μmol / l after feeding increased by 23% compared to the control (Fig. 5a).

In the RP-MHA feeding test, the taurine concentration of the control group was 49.78 ± 6.19 μmol / ℓ in the pre-feeding sample, and 49.78 ± 6.19, 42.49 ± 4.38, 42.78 ± 4.99 μmol / ℓ in the sample collected at two-week intervals after the feeding. After 6 weeks, it was 38.64 ± 1.63 μmol / ℓ. In the RP MHA matrix fed group, 46.95 ± 6.19μmol / ℓ before feeding and 51.79 ± 6.10, 63.32 ± 8.85 μ before sampling. Mol / l, 6 weeks after feeding, 59.19 ± 4.12 μmol / l significantly increased by 53% compared to the control (Fig. 5b).

An analysis of the yield and composition of crude oil samples did not show any significant increase or decrease in either the RP-Met Feed Test or the RP-MHA Feed Test. From the above results, when the precursor of taurine synthesis is added to the feed in the form of pellets or matrix resistant to the breakdown in the rumen, there is no change in the composition of milk protein or the production of crude oil, but the taurine content in milk is increased due to the increase of taurine biosynthesis in vivo. It was confirmed.

The method of producing milk with a high taurine content provided by the present invention prepares taurine precursors resistant to rumen degradation and supplies them with rich feeds to induce taurine synthesis in vivo, thereby artificially adding natural taurine or synthetic taurine. It can solve the problems of ruminant degradation and metabolic inhibition that occur during the production, and can produce taurine fortified milk with higher taurine content than ordinary milk, so it can be effectively used for the preparation of functional foods for infants requiring taurine supply. .

Claims (11)

1) preparing a precursor of taurine biosynthesis in the form of a protective amino acid that is resistant to rumen degradation; 2) feeding the feed containing the protective amino acid to raise livestock; And 3) A method of producing milk of a high taurine animal, comprising the step of harvesting crude oil from the livestock. The method of claim 1, wherein the precursor of taurine biosynthesis is methionine or methionine hydroxy analog. 3. The method of claim 2, wherein the methionine hydroxy analog is 2-hydroxy-4-methylthio butanoic acid. . 2. The protective amino acid according to claim 1, wherein the protective amino acid is resistant to rumen degradation and contains 60 to 70% by weight of taurine biosynthetic precursor methionine or methionine hydroxy analog as a core, and 20 to 25% by weight of insoluble fatty acid, starch, A method for producing milk of a high taurine content, characterized by pelletized from 2 to 10% by weight of dextrin or gelatin and from about 0.1 to 10% by weight of calcium hydroxide as a coating material. 5. The method of claim 4, wherein the milk of a high taurine content is characterized in that it comprises at least one component selected from the group comprising surfactants, antioxidants, antifoaming agents. 5. The method of claim 4, wherein the fatty acid is bee tallow. 7. The method of claim 6, wherein the tallow is hydrogen cured tallow. 2. The feeding of step 2 according to claim 1, wherein the feeding of the step 2 allows the protection amino acid pellets to be mixed with the rich feed and to consume the taurine precursor in an amount of about 0.1 to 1% of the daily feed, so that feeding is limited to livestock only during milking. Characterized in that, the production of milk of animal having a high taurine content. The method of claim 1, wherein the milk of the animal is milk obtained from cows, horses, goats, sheep or goats. Milk of taurine fortified animals produced by the method of any one of claims 1 to 10. A food or beverage composition comprising the taurine fortified milk of claim 10.