KR100493851B1 - Hydrolyzed yeast containing high concentration of selenium, preparing method thereof, and functional additives for livestock containing the same - Google Patents

Abstract

The present invention comprises the steps of culturing Saccharomyces cerevisiae (Saccharomyces cerevisiae) in a medium to which selenium salt is added, and then centrifuging to dry the precipitated cells to prepare selenium-containing yeast; Hydrolyzing the dry yeast under conditions of 5.0-7.5 pH and 50-70 ° C., by adding 0.5-2% by weight of protease to dry yeast to 5,000 daltons or less of yeast peptides; 75 to 80 mg of selenium salt (containing 30 to 40 mg of selenium) was added to 100 to 140 g of the hydrolyzed yeast peptide to react at 40 to 60 ° C. for 20 to 30 hours to obtain a selenium-binding yeast peptide, which was dried. It relates to a high concentration selenium-containing selenium yeast hydrolyzate prepared through a step, a preparation method thereof and a functional additive for livestock containing the same. High concentration selenium-containing yeast hydrolyzate according to the present invention is excellent in the anti-stress effect has the advantage of improving the productivity of livestock when used as a feed additive.

Description

Hydrolyzed yeast containing high concentration of selenium, preparing method etc, and functional additives for livestock containing the same}

The present invention relates to a high concentration of selenium-containing yeast hydrolyzate, a preparation method thereof, and a functional additive containing the same. More specifically, it contains a high concentration of organic selenium and has a high bioavailability, and has a molecular weight of 5,000 dalton or less, thereby easily passing the cerebrovascular barrier. It relates to a high concentration of selenium-containing yeast hydrolyzate having an anti-stress effect by affecting the neural tissue, a preparation method thereof, and a functional additive for livestock containing the same.

Mass mortality, growth, weakened immunity, reduced meat quality, and fatty liver disease caused by stress on livestock are economically significant losses. In particular, the suppression of pork PSE (Soft and Exudative) generation is a big concern for the pig industry, but there are no clear solutions to date. There is a situation. The occurrence of PSE in pork is most closely related to stress, which is the decisive factor in price deciding during pork rating. For example, assuming a loss of about 10,000 won per head, in 2001, a loss of W21.7bn (Monthly Pig Swine 2001, Korea Pig Association statistics) will occur for a 15% PSE incidence in 2001 on a 14,500 thousand slaughter basis.

Causes of stress on pigs include rapid temperature changes, high humidity, overcrowding, piglet types and locations, vaccinations, toxic gases, lack of feed and water, malnutrition, ventilation, disease, mycotoxins, and insect repellents. . When a pig is stressed, it causes physiological imbalances.For example, a temporary increase in serum corticosteroid levels inhibits the immune response, which increases body temperature, which leads to a loss of much fluid, which leads to electrolyte loss resulting in dehydration and diarrhea. Show symptoms. Diarrhea leads to an increase in feed demand, which results in a faster feed passage, resulting in a slower growth of pigs due to over-release of essential nutrients.

In addition, encroachment, poor ventilation, temperature, noise, parasites, disease and antibiotic administration put great stress on the most physiologically sensitive herds, causing chicken livers to become physiologically fatigued, causing fatty liver disease. The liver is an organ that plays a pivotal role in fat metabolism. When stress is stressed, fatty acid synthesis overloads fatty acids, which causes fatty acid transport mechanisms to supply various tissues in the body. Let's do it. More than 70% of domestic laying hens are known as fats, which accumulate excessive fat in the abdomen and press the egg ducts, causing fatty liver disease and fatty liver bleeding syndrome, which have recently caused economic damage to the poultry industry. Esau overlooked this and suffered huge economic losses.

As described above, the stress on livestock is causing huge economic losses to the livestock industry, but it is very difficult to create a breeding environment that can effectively suppress the occurrence of stress. Therefore, there is a need to develop a functional additive having an antistress effect that can be added to feeds that livestock encounter daily. Currently, feed additives that relieve stress from livestock have been underdeveloped. Existing products, which have synthetic vitamins and mineral minerals as their main ingredients, have lower bioavailability and less mitigative effects such as PSE pork. In addition, the development of anti-stress feed additives is insignificant, such as mineral and yeast supplements. Various trace minerals such as iron, calcium, and zinc are important components of animal body tissues and major enzymes. They perform important physiological functions such as immune function to disease and reinforcement of stress, but the density of breeding environment and excessive breeding As a result, the stressed livestock loses its ability to absorb stress, resulting in a lack of mineral absorption and excretion of a large amount of trace minerals in vitro, which leads to an increase in deficiency, causing disease and improving productivity.

Therefore, in order to produce and maintain fresh and high quality livestock products, hygiene management and stable specification management of livestock are required, while accumulation of residual substances, emergence of resistant bacteria, bacterial infection, and infection due to prolonged administration of antibiotics, sulfa drugs and pharmaceutical additives Increasing the immune response to side effects such as use, development of low-cost, high-quality anti-stress feed additives to solve urgent problems such as promoting fattening, increased fertility, and reduced mortality are required.

Therefore, in the present invention, when the inorganic selenium having an anti-stress effect is converted to organic selenium using yeast, and the yeast hydrolyzate treated with protease is ingested again with selenium and ingested to livestock, the above-mentioned problem is solved. It has been found that the present invention has been completed based on this.

Accordingly, it is an object of the present invention to provide a method for producing a high concentration of selenium-containing yeast hydrolyzate which is excellent in anti-stress effect of livestock and can improve livestock productivity.

Another object of the present invention is to provide a high concentration selenium-containing yeast hydrolyzate prepared by the above method.

Still another object of the present invention is to provide a functional additive for livestock containing the high concentration of selenium-containing yeast hydrolyzate.

Method for producing a high concentration of selenium-containing yeast hydrolyzate for the purpose of the present invention is Saccharomyces cerevisiae (Saccharomyces cerevisiae) incubated in a medium with the addition of selenium salt, followed by centrifugation to dry the precipitated cells to selenium-containing Preparing yeast; Hydrolyzing the dry yeast under conditions of 5.0-7.5 pH and 50-70 ° C., by adding 0.5-2% by weight of protease to dry yeast to 5,000 daltons or less of yeast peptides; 75 to 80 mg of selenium salt (containing 30 to 40 mg of selenium) was added to 100 to 140 g of the hydrolyzed yeast peptide to react at 40 to 60 ° C. for 20 to 30 hours to obtain a selenium-binding yeast peptide, which was dried. Steps.

The method may further comprise the step of removing off-flavors and already of the yeast hydrolyzate using activated carbon after the drying step.

High concentration selenium-containing yeast hydrolysates for other purposes of the present invention are prepared by the above method.

Functional additives for livestock for still another object of the present invention contains 0.01 to 0.1% by weight of the high concentration selenium-containing yeast hydrolyzate.

Hereinafter, the present invention will be described in more detail.

Selenium is a trace element distributed mainly in the liver, kidneys, heart and spleen in the human body. In the 1930's, animals fed on selenium-rich soils developed chronic food poisoning called alkaline disease, and research on selenium continued. The importance of this element in the human body was recognized in the late 1970s, and in the 1980s more information on the role of selenium led to the inclusion of selenium in the 10th revised nutritional recommendation in the United States. Selenium, which is also essential for humans, is an important component of glutathione peroxidase in red blood cells (an enzyme that breaks down hydrogen peroxide in the body and prevents damage to cells caused by hydrogen peroxide) and acts as an antioxidant like vitamin E. Compounds such as methionine and selenocysteine have been known to have antioxidant, radiation defense, and anticancer effects, but recently, selenium has been shown to have antidepressant effects, so that selenium acts on the hypothalamus-pituitary-nerve system. , Anti-anxiety, etc. (Biol Psychiatry 1991 Jun. 1; 29 (11) 1092-8: The impact of selenium supplementation on mood).

Organic selenium is 5 times more bioavailable than inorganic selenium, and yeast-derived low molecular weight peptides with molecular weight less than 5,000 daltons are formed to prevent the entry of macromolecules into the blood vessels of the brain to protect brain cells. It is expected that the anti-stress ripple effect of the selenium-binding yeast peptide will be very large because it can easily pass through the organs and only low molecular weight substances can enter the cerebrovascular vessels, affecting the neural tissue and thus having an excellent anti-stress effect. .

Selenium can be absorbed in the form of organic selenium and inorganic selenium. The organic selenium has a much higher absorption rate in the body than inorganic selenium. Inorganic selenium has a very poor absorption in the body and forms complexes with other minerals and insoluble forms. Therefore, it is necessary to increase the bioavailability by increasing the organic passivation rate of selenium, and side effects such as disorders of the digestive organs, jaundice, and hair loss may occur when selenium is excessively ingested. . In addition, in the livestock, various nervous system abnormalities such as mad cow disease are induced, but the only solution to this is through mineral supplementation. Therefore, we developed a material that can act as a neurotrophic ingredient, neurotransmitter, or neurotransmitter precursor. If it is solved by various types of neurological diseases and stress caused by feeding in the form of additives, the market as a functional feed additive can be considered infinite.

According to the present invention, the conventional organic selenium yeast is only used as a source of selenium, but only the effects of promoting sperm production and simple meat enhancement, etc., by converting it into the organic selenium chelated peptide form can obtain a strong anti-stress effect have. The chelate is covalently bonded with amino or oxygen residues to form a cyclic structure by combining with minerals, so the bioavailability is significantly higher than that of inorganic minerals, and it does not form an insoluble substance by combining with other nutrients in the intestine. Also can exhibit a high effect.

Selenium-binding yeast peptides produced by low molecular peptides of organic forms of selenium-binding yeast proteins can take advantage of the anti-stress and antidepressant effects of selenium and can be fed in the form of organic selenium rather than inorganic selenium. ) Can be maximized. In addition, it can easily pass through the cerebrovascular barrier by low molecular peptide, can act as a neurotrophic component and have an antistress function, and can act as a neurotrophic factor, a neurotransmitter precursor of yeast-derived peptides can have an antistress effect. In addition, even a small amount of 0.01-0.1% by weight of the blending ratio per feed has a suitable anti-stress effect.

According to one embodiment of the present invention, Saccharomyces cerevisiae is incubated for 24 hours as a strain for producing high concentration selenium-containing yeast hydrolyzate. The culture solution having the cultured solid content of 10-15% was inoculated in a medium sterilized at 121 ° C. for 15 minutes by adding urea, ammonium sulfate, molasses, phosphoric acid, potassium chloride, magnesium sulfate, calcium sulfate, zinc sulfate, and thiamine in distilled water. .

The inoculated strain was incubated for 17-18 hours in 25-35 ° C., 5.0-6.0 pH and aerobic conditions in a 1 liter culture flask, at which the selenium concentration was 1000-1200 ppm at the time when the growth of the yeast was stopped. Incubate by adding selenium salt if possible. If the concentration of selenium to be added is less than the above range, the organic passivation rate of selenium is high, but there is a problem of low economic feasibility, if it exceeds, there is a disadvantage that the organic passivation rate of selenium falls.

Subsequently, the culture solution is centrifuged at 15,000 rpm for 15 minutes to remove the supernatant, and the selenium-containing yeast, which is a precipitated cell, is washed with distilled water and dried to prepare a selenium-containing yeast.

In order to decompose the yeast into a yeast peptide containing 75% or more of a peptide having a molecular weight of 5000 dalton or less, by adding 0.5 to 2% by weight of protease to dry yeast under conditions of 5.0 to 7.5 pH and 50 to 70 ℃ Hydrolysis for 24 hours. As the protease for hydrolysis, papain-based protease is preferred.

The molecular weight of the hydrolyzed peptide is preferably 5,000 dalton or less, and if it exceeds 5000 dalton, the cerebrovascular barrier is not easy to pass and affects the neural tissues and thus cannot act as an antistress.

To convert the hydrolyzed yeast peptide into a high concentration of selenium-containing yeast peptide, 75 to 80 mg (containing 30 to 40 mg of selenium) of selenium salt was added to 100 to 140 g of the hydrolyzed yeast peptide, followed by 40 to 60 ° C. Reaction is carried out at a high temperature of 20 to 30 hours, and dried to prepare a high concentration selenium-containing yeast hydrolyzate. If the added amount of selenium salt is less than the above range, the organic passivation rate of selenium is high, but there is a problem of low economic efficiency, if it exceeds, there is a disadvantage that the organic passivation rate of selenium falls.

The reaction scheme where selenium binds to the hydrolyzed yeast peptide is shown in Scheme 1 below.

Na ₂ SeO ₃ → 2Na ⁺ + SeO ₃ ^2- → SeO ₃ ^2- + 3H ₂ → Se ⁺⁴ + 3H ₂ O

Se ⁺⁴ + 4 [amino acid (peptide)] ^- → Se-4 [amino acid (peptide)]

According to the above scheme, selenium-containing hydrolyzate to which selenium is bound is prepared.

On the other hand, it can optionally remove off-flavor and odor of the hydrolyzate prepared above. Off-flavor and already has the disadvantage of degrading the quality of the product, it is desirable to remove it, purification treatment using activated carbon is common.

High concentration selenium-containing yeast hydrolyzate prepared according to the present invention may be included in the feed for livestock in an amount of 0.01 to 0.1% by weight, if the addition amount is less than the above range there is a disadvantage that the desired anti-stress effect can not be obtained, Since side effects such as disorders of the digestive organs, jaundice, and hair loss may occur, an appropriate feed amount of selenium is preferably 0.01 to 0.1% by weight as a functional additive for livestock.

As described above, the highly concentrated selenium-containing yeast hydrolyzate prepared according to the present invention is not inorganic selenium, but an organic selenium having a high absorption rate in the body, and thus has a molecular weight of 5000 dalton or less, so that the selenium is easily passed through the cerebrovascular barrier. By affecting the tissue, it is possible to obtain the anti-stress effect of selenium, and when included in the animal feed, there is an advantage to improve the productivity by reducing the stress of the livestock. In addition, the method for producing high concentration selenium-containing yeast hydrolyzate is easy, which is very advantageous and economical for mass production.

Hereinafter, the present invention will be described in more detail with reference to the following Examples, but the scope of the present invention is not limited to the following Examples.

Example 1

Preparation of high concentration selenium-containing yeast hydrolyzate

Saccharomyces cerevisiae strain (KCTC 7904) was incubated for 24 hours, and 10 ml of the culture solution having a solid content of 20% was added to 1 liter of distilled water, 3.08 g of urea, 0.97 g of ammonium sulfate, 88.2 g of phosphoric acid, 0.73 g of phosphoric acid, 277 mg of potassium chloride, 214 mg of magnesium sulfate, 143 mg of calcium sulfate, 7.96 mg of zinc sulfate, and 1.07 mg of thiamin were added thereto and inoculated in a medium sterilized at 121 ° C. for 15 minutes. The inoculated strains were incubated in nine 1 liter flasks for 17.5 hours at 30 ° C., 5.0-6.0 pH and aerobic conditions, respectively, to prepare nine samples. After 8 hours of incubation, selenium concentration was 74 as shown in Table 2 below. Selenium sodium (sodium selenite) was added to incubate at ˜4,791 ppm. The nine cultures were centrifuged at 15,000 rpm for 15 minutes to remove the supernatant, and the selenium-containing yeast, which was precipitated cells, was washed with distilled water and dried to prepare yeast-containing yeast.

In order to analyze the yield of the selenium-containing yeast, after centrifugation (5,000 g, 15 minutes), selenium-containing yeast was recovered and dried to determine the weight. To measure selenium content in yeast, 0.1 g of dry weight was added to a 50 ml vial and diluted by acid decomposition (consisting of 30% nitric acid, 20% hydrogen peroxide, and 50% distilled water). After heating for 2 hours to obtain a clear solution, the solution was introduced into ICP-AES (Inductively Coupled Plasma-Atomic Emission Spectrometry) to measure the concentration of selenium.

In order to measure the organic form of selenium, 0.1 gram of MBRT (Methylene Blue Reduction Time) method, which is a method of Ali Demirci (1999) measurement, dried sample was reduced solution (20% (W / V) solution; 0.2 N To 7 ml of a solution containing 20% (w / v) 1-thioglycerol in a phosphate buffer solution, pH 5.5), the solution was stirred vigorously at 200 rpm for 3 minutes in a sealed state, and a drop of 2% methylene blue solution was dropped. By measuring the disappearance of the blue color, the method of measuring the content by analyzing the presence characteristics of the mineral in the organic form was used, and the results are shown in Table 1 below. The yield, selenium concentration and MBRT measured for the selenium-containing yeast are shown in Table 2 below.

                MBRT mineral concentration 0 0.5-7 15-20 20 or more ppm More than 10ppm 10 ppm Extremely low 0

Sample number Dose of sodium selenium (mg) Incubation time (h) Dry yeast yield (g) Total selenium content (ppm) MBRT (minutes) One 0 17.5 11.2 74 20> 2 98.55 17.5 10.9 1074 20 3 219 17.5 10.2 2376 3 4 328.5 17.5 10.0 2514 3 5 438 17.5 9.8 2603 3 6 547.5 17.5 9.6 2712 2 7 657 17.5 9.2 3606 2 8 876 17.5 7.8 4393 One 9 1095 17.5 7.2 4791 One

As shown in Table 2, when the selenium sodium of 98.55mg level was added, the yield of yeast, selenium content in dried yeast, and MBRT showed the best results. Here, yeast production was only 2.7% different from sample 1 containing no selenium sodium, the selenium concentration in yeast was 1074ppm, and MBRT was more than 20 minutes, indicating that most of the selenium is present in yeast in organic form. there was. As the concentration of inorganic selenium increased, the selenium content of yeast increased, but in most cases, MBRT appeared within 1 to 3 minutes, indicating that the selenium was present in inorganic form.

In order to decompose the yeast of Sample No. 2, which was excellent in the above test results, into peptides, autodigestion was performed at 6.0 pH and 55 ° C., and a collupurin MG (DSM Co., Ltd.) having a unit of 30,000 IU was applied to the dry yeast. 1% was added and hydrolyzed for 24 hours. 76.65 mg of selenium salt having a selenium content of 35 mg relative to the dry yeast weight ratio was added to the hydrolyzed yeast peptide, and then reacted at a high temperature of 50 ° C. for 24 hours, and dried to prepare a high concentration selenium-containing yeast hydrolyzate. The hydrolyzed peptide was measured for molecular weight distribution using UF (Ultrafiltration, Ultra Filtration.) Method. In addition, the yield, selenium concentration and MBRT of the hydrolyzate were measured in the same manner as above and shown in Table 3.

TABLE 3

Sample number Selenium Sodium Input Reaction time Yield Total Selenium Concentration MBRT 2 76.65mg 24 hours 11.12 g 4,199 ppm More than 15min

Comparative Example 1 and Examples 2 to 4

The high concentration selenium-containing yeast hydrolyzate prepared according to the present invention was treated with five treatment groups, namely, control group 1 (25 days crumble salary → 8 days crumble salary), Comparative Example 1 (25 days crumble salary → 8 days pellet salary), Example 2 (25 days crumble feed → 8 days pellet + selenium-containing yeast hydrolyzate 0.05% feed), Example 3 (25 days crumble feed → 8 days pellet + selenium-containing yeast hydrolyzate feed 0.10%) and Example It was fed to broiler chickens by dividing it into 4 (25 days of crumble feeding → 8 days of pellet + 0.2% of yeast hydrolyzate containing selenium). Stress was measured. Evaluation of this was determined by weight gain, and the results are shown in Table 4 below.

division Weight gain before feed change Weight gain after feed change Control group 1 49.49 g / day 49.18 g / day Comparative Example 1 48.79 g / day 47.58 g / day Example 2 48.45 g / day 50.61 g / day Example 3 48.75 g / day 51.21 g / day Example 4 48.63 g / day 50.46 g / day

As can be seen in Table 4, the weight is reduced when stress is applied, but the weight gain was significantly increased upon administration of the selenium-containing yeast hydrolyzate of the present invention.

In addition, when the selenium-containing yeast hydrolyzate of the present invention is added at 0.2% or more, it can be seen that the weight gain rate is reduced.

Comparative Example 2 and Example 5

For 24 pigs 5 days prior to release, 12 feeds were fed with 0.05% selenium-containing yeast hydrolyzate to the feed (Example 5) and 12 feeds were not added (Comparative Example 2). As a result, the average live weight of the example was measured at 121 kg, and the comparative example was measured at 112 kg. In addition, the PSE of the examples was significantly reduced (see Table 5).

Example 5 Comparative Example 2 Conductor Number Meat color Conductor Number Meat color One 3 13 3 2 4 14 2 3 3 15 2 4 4 16 2 5 4 17 2 6 3 18 2 7 4 19 2 8 3 20 2 9 3 21 2 10 3 22 2 11 3 23 2 12 4 24 3  1: light color-PSE meat (abnormal meat) 2: slightly light color-PSE meat (abnormal meat) 3: normal color-normal meat 4: slightly dark-normal meat 5: dark-normal meat 6: very dark -DFD meat (abnormal meat)

Comparative Example 3 and Example 6

Restraint stress experiments according to Brekhman and Dardymov method

After selenium-containing yeast hydrolyzate of the present invention was orally administered for 5 consecutive days, the rats were fixed to 45% of ramps at 3 days, and after 48 hours of stress induction, the adrenal glands, thymus, spleen and thyroid gland were extracted and their weights. Example 6, which was measured, and Comparative Example 3, which did not receive the selenium-containing yeast hydrolyzate, and the control group that were not stressed were compared. As a result, Comparative Example 3 showed a weight change in which the spleen was reduced and the kidney and the thymus were enlarged. Example 6 showed the weight close to the control group that was not stressed even when stress was applied (see FIG. 1). In addition, as a result of measuring the blood stress-related enzyme concentration, such as LDH in the serum of the rat, Example 6 showed a blood enzyme concentration close to the normal group even when stress is applied (see Fig. 2).

As described above, the high concentration of selenium-containing yeast hydrolyzate prepared according to the present invention is a substance in which selenium is converted into an organic state and has a low molecular weight of 5,000 dalton or less, and thus has high bioavailability of selenium and easily passes through the cerebrovascular barrier. By affecting the nervous system is excellent anti-stress effect.

In addition, as can be seen through the above embodiment, the high concentration of selenium-containing yeast hydrolyzate of the present invention improved the feed efficiency of broiler, and in the case of pigs, the PSE of the piglet was reduced to improve meat quality. In addition, it is expected that in cattle, it may be possible to alleviate feed stress and calf movement stress, to improve eggshell egg laying, and to reduce weaning stress of pigs. In addition, it can be developed as a fish feed, and anti-stress preparations for dogs are expected to reduce the incidence of disease caused by stress. Therefore, when the high concentration of selenium-containing yeast hydrolyzate of the present invention is added to livestock feed and ingested to livestock, it shows an excellent stress suppressing effect compared to the existing feed, thereby improving the meat quality, breeding rate and growth rate of livestock industry, thereby improving productivity of the livestock industry. Will contribute greatly. In addition, by using this as a natural substitute for antibiotics can help reduce the accumulation of antibiotics in the meat of the meat, it is expected to play a major role in the prevention of mad cow disease by developing as a neurological strengthening agent of ruminants as a neuroactive substance and neuronutrients. In addition, by incorporating into the food industry, it will be possible to develop high-purity selenium anticancer food.

1 is a graph showing the results of restraint stress experiments (weight measurement of adrenal gland, thymus, spleen and thyroid gland, etc.) according to Brekhman and Dardymov method according to Example 6 and Comparative Example 3 of the present invention to be.

2 is a graph showing the results of restraint stress experiments (measurement of blood enzyme concentration) according to the Brekhman and Dardymov method according to Example 6 and Comparative Example 3 of the present invention.

Claims (5)

Saccharomyces cerevisiae (Saccharomyces cerevisiae) incubated in a medium to which selenium salt was added, and then centrifuged to dry the precipitated cells to prepare a selenium-containing yeast; Hydrolyzing the dry yeast under conditions of 5.0-7.5 pH and 50-70 ° C., by adding 0.5-2% by weight of protease to dry yeast to 5,000 daltons or less of yeast peptides; And 75 to 80 mg of selenium salt (containing 30 to 40 mg of selenium) was added to 100 to 140 g of the hydrolyzed yeast peptide to react at 40 to 60 ° C. for 20 to 30 hours to obtain a selenium-binding yeast peptide, which was dried. Method for producing a high concentration selenium-containing yeast hydrolyzate, comprising the step of. The method according to claim 1, wherein the method further comprises the step of removing off-flavor and yeast of the yeast hydrolyzate using activated carbon after the drying step. The method of claim 1, wherein the selenium-binding yeast peptide contains 75% or more of a peptide having a molecular weight of 5,000 dalton or less. High concentration selenium-containing yeast hydrolyzate prepared by the method of any one of claims 1 to 3. A functional additive for livestock, comprising the high concentration of selenium-containing yeast hydrolyzate of claim 4 in an amount of 0.01 to 0.1% by weight.