KR20040040222A - Yeast hydrolysate comprising Angelica Gigantis Nakai having anti-stress, anti-fatigue, and neurotrophic activities - Google Patents

Abstract

PURPOSE: Provided is a method for manufacturing a yeast hydrolysate with anti-stress, anti-fatigue, and neurotrophic activities containing Angelica Gigantis Nakai, thereby increasing anti-stress, anti-fatigue, and neurotrophic activities thereof more. CONSTITUTION: A method for manufacturing a yeast hydrolysate with anti-stress, anti-fatigue, and neurotrophic activities comprises the steps of culturing yeast strains and adding the extract of Angelica Gigantis Nakai, to the cultured yeast strains, followed by autolysis at 35-70 deg.C.

Description

Yeast hydrolysate comprising Angelica Gigantis Nakai having anti-stress, anti-fatigue, and neurotrophic activities

The present invention relates to a method for producing yeast hydrolyzate (ADNF: Angellica Gigantis Nakai Drived Neurotrophic Factor) having improved antistress and anti-fatigue activity, and more particularly, to the hydrolysis of yeast during the production process of yeast hydrolyzate. By adding the Angelica extract at the time of aging or self-extinguishing, it provides a method for producing a yeast hydrolyzate including Angelica extract, which has an improved anti-stress and anti-fatigue effect.

In general, stress is an environmental, mental, or physical stimulus that affects a living body, and the living body responds to various stimuli to maintain homeostasis. In other words, stress acts on the central nervous system (Glavin et al., 1985), including the immune system (David et al., 1990), the digestive system, and the cardiovascular system (Selye, 1983). Although it causes general adaptation syndrome (Selye, 1958), it is a bioregulatory process for maintaining homeostasis against environmental changes.

The mechanism of stress and its effect on the body are not yet known, but stress response is mainly through the hypothalamus-pituitary-adrenal system (Selye, 1950; Chowers et al., 1967; Hodges et al., 1976; Laural). Et al., 1989) are intricately interrelated and affect each other. External stimulation affects the limbic system of the cerebrum, which stimulates the secretion of ACTH from the pituitary gland, which stimulates the adrenal cortex, facilitating the consumption of ascorbic acid and the biosynthesis of corticosteroids. 1936)

Recently, studies on drugs to suppress such stress have been actively conducted, among which urusdeoxycholine acid, which is widely used for liver disease, inhibits stress-induced ulcers and lipid peroxidation of gastric mucosa. Kawamura et al. 1989: Cho et al., 1995)

In modern society, science and technology are advanced and complicated, and social situation is volatile. People are exposed to various stresses. Especially in the internationalized society, complicated relationships are formed and various symptoms caused by mental stress have been reported.

Mental stress is said to have a big impact on the circulatory system and immune system. For example, stress has been reported to increase angiotensin II and the like, and excess sodium in the body due to sodium resorption causes blood pressure to increase (Osamu Shigehara et al .: Metabolism, 28, 2, 323, 1991). On the basis of this, the effects of the angiotensin converting enzyme inhibitors enalapril and alasepril, which are used as antihypertensive drugs, on the hypertension caused by stress have been studied (The American Journal of Cardiology; 68, 15, 1362 (1991), Internal Medicine; 32, 9, 691 (1993)). However, stress load not only causes an increase in blood pressure, but also influences various factors, and is thought to be a cause of not only hypertension but also peptic ulcer, ischemic heart disease, cerebrovascular disorder, and hyperlipidemia. Therefore, although stress is considered to be one of the causes of hypertension, on the contrary, it is not considered that antistress effect is obtained by simply suppressing blood pressure rise.

Currently, chemical synthesis agents such as mental stabilizers, anti-anxiety agents, and sleeping pills are used as agents for reducing and preventing mental and physical symptoms caused by stress. However, these drugs have problems of habituality and side effects, so it is not desirable to use these drugs routinely for the purpose of preventing mental and physical symptoms caused by stress. Therefore, there is a demand for an antistress agent that can be used in daily life and that does not include safety problems and exhibits an effect of reducing and preventing mental and physical symptoms caused by stress. For example, an antistress agent containing L-theanine contained in tea leaves (see Japanese Patent Application Laid-Open No. 6-100442), an antiseptic compound containing an imidazole compound such as anserine, valenin, n-methylhistidine, τ-methylhistidine, etc. A stress composition (Japanese Patent Laid-Open No. Hei 9-20660), and a stress improving food containing a composition of glutathione and an antioxidant (Japanese Patent Laid-Open No. Hei 8-275752) have been proposed. Also, FRAGRANCE JOURNAL: 1991-11, p44-49 has been reported on the effect of fragrance stress.

In addition, anti-stress agents having an effect on the prevention and treatment of stress using choline acids have been developed (Korean Patent No. 95-6225). The antistress agent containing choline acids is widely used as a gallstone dissolving agent, and has a dipharyngeal effect, liver detoxification, hepatic blood flow improving action, fat absorption promoting action, and excretion of waste products through microbilitery. It is marketed as urusdeoxycholic acid, a gallstone dissolving agent, which is known to have an effect on improvement of function, systemic malaise due to liver dysfunction, indigestion, anorexia, physical fatigue, and it is effective in treating fatty liver. When administered orally, hepatofunctional improvement and hepatic fat loss have been reported (see Japanese Patent Application Laid-Open No. Hei 4-235918). Taurorsodeoxycholic acid, chenodeoxycholine sold as a gallstone dissolving agent Acid (Chenodeoxycholic acid) and dihydrocholic acid (Dehydrocholic acid) contains as an active ingredient.

However, the anti-stress composition using choline acids is effective in the general symptoms caused by stress, such as increase in adrenal weight caused by stress and prevention of spleen atrophy, suppression of ascorbic acid content change of adrenal gland and blood biochemical change. It has been demonstrated that there is no mention of the effects of suppressing or restoring memory loss.

In addition, various types of medicines have been developed (Korean Patent Application No. 96-8230), which contains ingredients such as ginseng, tofu, and thorny ginseng, which are known to have a beneficial effect on stress as a herbal or folk remedy. Nutrient tonic drinks are mainly marketed as a main ingredient and are also commercially available to drink the main ingredient of two worms, and various other ingredients such as honey and vitamins are also marketed as a nourishment tonic. However, Ginseng, B. g. (Brekhman, II, Pharmacological investigation of glycosides from Gingseng and Eleutherococcus, Lloydia, Vol. 32, No. 1, pp. 46-51, 1969), Ocimum sanctum Linn, Tinosporamalabarica Herbs and melatonin, such as (Sen, P., Maiti, PC and Ray, A. Mechanism of anti-stress activity of Linn, eugenol and in experimentalanimals, Indian J. Exp. Biol. , 30, 592-596, 1992) Although it has been reported that the biomaterial has an anti-stress action, only a few of the various changes caused by stress appear to be effective or ineffective, and simply use herbal extracts that have been used as herbal medicines in the Orient. In this respect, a large characteristic technology was not found, and the general purpose was merely to use nourishment tonic or known efficacy.

Accordingly, the present invention provides a novel neurotrophic substance having an excellent effect in the treatment of neurological diseases including autonomic nervous system diseases by having anti-stress, anti-anxiety, anti-fatigue, nerve stabilizing and sleep assisting activity without side effects. The purpose.

In order to achieve the above object, the present invention,

Culturing the yeast strain to the maximum growth phase; And adding the donkey extract to the cultured yeast and self-extinguishing at a temperature of 35 ° C. to 70 ° C .; comprising a, antistress and anti-fatigue function. To provide. If the temperature is less than 35 ℃ self-digestion is not well progressed, if more than 70 ℃ will proceed to kill the cells.

In this process, the fermentation of the Angelica extract naturally proceeds, it is possible to obtain a more excellent effect of Angelica fermentation.

At this time, at the same time as or after autodigestion, hydrolysis can be further performed by further adding a proteolytic enzyme.

In the above method, the self-digested yeast hydrolyzate may further include the step of centrifugation to separate only the supernatant. Preferably, the supernatant taken by centrifugation is filtered by using an ultrafiltration membrane for ultrafiltration. The method may further include taking only a peptide having a molecular weight of 10,000 or less.

In the above method, the Angelica extract can be obtained by heating and extracting Angelica with 40 to 100 ° C. of ethanol, followed by concentration under reduced pressure and drying, and the Angelica extract is preferably added in an amount of 0.1 to 30% by weight. . If the Angelica extract is less than 0.1%, the effect is insignificant, if more than 30% by weight can not obtain the effect of the desired yeast extract.

In the above method, preferably, the step of improving the yeast strain by selecting only the surviving strains by culturing while applying the high temperature, ultrasonic wave, vibration, pH change, etc. in the range that yeast does not die before cultivating the yeast to the maximum growth stage It may further include. This improved yeast strain has a greater ability to resist stress.

In the above method, it is preferable to induce the secretion of metabolites to withstand stress by applying appropriate physical or chemical stress such as high temperature, ultrasonic wave, vibration, pH change, etc. in the range that yeast does not die before the yeast self-extinguishment. It may further comprise a step. In response to this stress, yeast is able to more actively secrete antistress substances.

The present invention further provides an antistress agent comprising a yeast hydrolyzate comprising the fermented Angelica extract prepared as described above as an active ingredient. At this time, the anti-stress agent may have activity of nerve stabilization and tension relaxation.

The present invention further provides a fatigue recovery formulation comprising the yeast hydrolyzate comprising the fermented Angelica extract, prepared by the above method, as an active ingredient.

The present invention further provides a neuronutrient comprising the yeast hydrolyzate comprising the fermented Angelica extract prepared according to the above method as an active ingredient.

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

The present invention provides a method for preparing a yeast extract having anti-stress and anti-fatigue activity, wherein the yeast extract is prepared by self-extinguishing the yeast, wherein the Angelica extract is added during the self-extinguishing process. In the present invention, the method for preparing the basic yeast extract used the method for preparing the yeast extract described in Patent Application No. 01-9946. According to the production method of the present invention, it is possible to obtain a yeast extract having a superior anti-stress and anti-fatigue effect than the yeast-derived peptide according to the patent application No. 01-9946.

The addition of Angelica extract during the hydrolysis of yeast was conceived to increase the bioavailability of the human body by decomposing and aging the active ingredient of Angelica easily to be digested and absorbed by the human body using the autosomal digestive enzyme of yeast. When aging ginseng to take the effect is excellent (red ginseng) and the fermentation of various herbal ingredients to prepare a liquor, the present invention was completed by considering the fact that its health promoting function can be excellent.

Yeast has a self-extinguishing function, and if it is in an environment where it is far out of conditions (temperature, pH, etc.) where it can grow or in starvation without nutrients, the cytoplasm of the yeast is hydrolyzed by the action of its own enzyme. At this time, the cell wall is also destroyed by the continuous enzymatic action, and the already decomposed various cytoplasm is eluted out of the cell through the broken cell wall to increase the bioavailability of the yeast. By dissolving the various proteins, minerals, vitamin B group, etc. contained in the yeast to act as a more efficient neurotrophic substance to obtain a yeast hydrolysate having an anti-stress effect, and extracts Angelica extract with a calming function When hydrolyzed with yeast's auto-digesting enzyme or protease, it is more anti-stress It is possible to make a formulation having excellent anti-fatigue effect.

In view of this, the present invention is to self-digest the yeast bacteria at high temperature of 35 ℃ to 70 ℃, and then auto-digested together, and further hydrolyzed by the addition of proteolytic enzyme, and the hydrolyzed The Angelica fermentation hydrolyzate containing the Angelica hydrolyzate and yeast hydrolyzate was prepared by centrifuging and separating only the supernatant.

In order to test whether the anti-stress capacity through the neurotrophic function of the Angelica fermentation yeast hydrolyzate of the present invention, as the test method below, Brekman and Darmov (Brekhman) and Dardymov) were applied to the floating stress experiments.

In general, various reactions of the body caused by stress are initiated through the detection of external stimuli of the central nervous system, and in response to these external stimuli, the corticosteroid secretion is changed and the liver, thymus, and thyroid gland are And changes in the weight of the spleen and the like, resulting in a decrease in immune cells, and changes in blood lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) enzymes under the influence of corticosteroids. (Christina, J., Hans, W. and Hans, M .: Haematological changes during acute mental stress, Br. J. Heanat 71: 564, 1971 / Conner, RL, Vernikos-Danelis, J. and Levine, S .: Stress, fighting and neuroendocrine function, Nature, 2: 564, 1971./Munck, A., Guyre, PM and Holbrook, NJ: Physiological functions of glucocorticoids in stress and their relation to pharmacological actions.Endocrine Reviews., 5:25, 1984.) The change was measured and found to have an antistress effect.

In addition, as an anti-stress test of the present invention, the anti-fatigue effect (anti-fatigue capacity) experiment was performed by measuring the swimming exercise ability of the mouse.

It is known that stress affects affective reaction, motor function, and autonomic function (Hwang, Ee-Wan, 心身 症, Seoul, Haenglim Publishing Co., pp.17-28, 33-49, pp.277, 1985), In general, excessive exercise (swimming) is regarded as stress, and the anti-fatigue effect through the measurement of swimming endurance capacity, which measures the ability to last long while being forced to swim excessively through a swimming pool measuring device. Experiments are a good way to measure antistress effects. As a result of the experiment, it was confirmed that the present invention Angelica fermentation yeast hydrolyzate has better anti-stress effect and anti-fatigue effect than general Angelica extract and yeast hydrolyzate.

Example

The present invention will be described in more detail with reference to the following Examples, which should not be construed as limiting the invention, but may be modified by those skilled in the art within the scope of the invention.

Example 1

Preparation of Angelica Extract and Angelica Fermented Yeast Hydrolyzate

The Korean Angelica used in this Example was used as a quality commodity at Gyeongdong Market Herbal Medicines Wholesaler to chop the Angelica into fine powder, and extract it by heating it at about 70 ℃ three times for 2 hours with ethanol, and filtrate to concentrate the filtrate under reduced pressure. The soft extract was made and dried again in a desiccator. The yield was 44.26%, as 221.28 g per 500.0 g of Angelica.

Food grade yeast Saccharomyces cerevisiae strain (purchased by Genico Co., Ltd.) was incubated in YM medium at 24 ° C. and oxygen for 48 hours.

After centrifuging the culture broth as described above for 15 minutes at 15,000 rpm, the supernatant was removed, and the remaining precipitate, ie, the yeast was washed and diluted twice with 10 times (v / v) sterilized water, and then the yeast was heated at a high temperature of 50 ° C. In addition to self digestion using an autolysis enzyme that dissolves itself, the Angelica extract is added in about 10% by weight of yeast, and the protease (papaine 30.000) is promoted to promote the breakdown of the yeast protein into peptides. ) Was added to 1% and hydrolysis was performed for 48 hours at initial pH 4.0 to dissolve the protein as peptide. Thus dissolved was centrifuged at 15,000 rpm to separate only the supernatant in aqueous solution to obtain the Angelica fermentation yeast hydrolyzate of the present invention.

The lyophilized product of the yeast peptide containing the Angelica fermentation hydrolyzate thus obtained was called "Sample 1" and used in the following experimental example.

Example 2

1) Selection of improved strains of yeast

The Saccharomyces cerevisiae7904 strain, a food-grade Saccharomyces cerevisiae7904, sold by the Biotechnology Research Institute (KCTC), was incubated for 48 hours under stress conditions subject to vibration using a continuous drop of 45 kHz and a submersible pump. Yeast was screened on YM agar plates (3g yeast extract, 3g malt extract, 5g peptone, 10g glucose, 15g / L) to select the fastest growing and large colonies on the plate as an improved strain.

2) Culture

The selected strains were incubated for 48 hours in a fermenter at 24 ° C. and oxygen supply in YM medium, and the culture solution was centrifuged at 15,000 rpm for 15 minutes to remove the supernatant. After diluting 10 times with the buffer solution, the process of adding high temperature of 35 ℃ to 45 ℃ and 45kHz ultrasonic wave for 5 seconds at 30 minutes in a fermentation tank equipped with ultrasonic generator and vibrator was repeated for 8 hours. During the 8 hours incubation while giving a vibration using a drop to produce a lot of anti-stress substances secreted by the yeast due to stress.

3) Autodigestion and Hydrolysis

The culture cultured in the above 2) was subjected to self-extinguishing using a self-extinguishing enzyme in which yeast dissolves itself at a high temperature of 50 ° C. In addition, about 10% of the Angelica extract was added on the basis of autodigestion, 1% of protease (papaine 30.000) was added to promote the hydrolysis of the yeast protein into peptides, and hydrolysis was performed for 48 hours at an initial pH of 4.0. Protein was dissolved with peptide.

Hydrolysis rate (%) after degradation was determined as the protein concentration present in the supernatant divided by the total yeast protein concentration. The protein concentration was centrifuged to recover the supernatant by centrifuging the autodigested sample, and 500 µl of the diluted solution was added to the tube, the same amount of protein quantitative reagent was added, stirred, and reacted at 60 ° C. for 60 minutes. Measured at 562 nm. As a result, the hydrolysis rate was 57%.

4) Separation, Purification and Drying of Hydrolysates

As described in 3), the digested and hydrolyzed one was centrifuged at 15,000 rpm to separate and obtain only the supernatant solution in an aqueous state, which was used in the following Experimental Example as "Sample 2," and the precipitate as "Sample 3". It was also used in the following experimental example. The supernatant and the precipitate thus obtained were lyophilized to obtain a powdered Angelica fermented yeast extract and precipitate.

Example 3

The yeast extract was prepared in the same manner as in Example 2, but the obtained supernatant was separated and purified through an ultrafiltration membrane (MWCO = 10,000) through an ultrafiltration device, and then lyophilized to be purified to a molecular weight of 10,000 or less. Natural yeast peptides were prepared.

Example 4

Amino acid analysis

10 g of the Angelica fermented yeast extract obtained in Example 2 was dehydrated with cold acetone, and then dried on a filter paper in a 60 ° C. dry oven, and then 5 mg of the sample was added to a hard test tube, and 5 ml of 6 N HCl was degassed and sealed. Hydrolyze at 110 ° C for 24 hours, wash with three small portions of distilled water, dry at 50 ° C with concentrator to remove hydrochloric acid, dissolve buffer, and exchange column Li 10cm No.338051 with amino acid automatic analyzer (Deckman system 6300, USA) It was analyzed using. As a result of analyzing the amino acid composition, the amino acid composition of the yeast extract of the present invention was as shown in Table 1 below.

amino acid Molar ratio (%) Glutamic acid 14.5 Leucine 5.8 Lysine 4.8 Aspartic acid 5.9 Alanine 4.1 Histidine 4.3 Isoleucine 3.2 Phenylalanine 4.9 Valine 3.4 Tyrosine 4.6 Threonine 3.0 Glycine 3.0 Serine 2.8 Arginine 2.3 Proline 3.5 Tryptophan 2.8 Methionine 1.4 Cysteine 0.9

As shown in Table 1, the yeast peptide added to the Angelica extract prepared in the present invention contains 18 different amino acids, in particular, the content of aspartic acid and glutamic acid is very high, and patent application 01-9946 It can be seen that the content of phenylalanine, tyrosine, proline, and tryptophan increased compared to the yeast extract described in the above. It appears to be due to coumarin, decusin, and decosinol among the essential oils of Angelica extract.

As can be seen from the above, various amino acids and peptides can be usefully used as anti-stress substances that can act as neurotrophic substances and control the imbalance of autonomic nerves generated during excessive stress.

Hereinafter, the anti-stress activity, anti-fatigue activity and the like of the yeast peptides prepared by the Angelica extract fermentation of the present invention were tested.

Experimental Example 1

Antistress Activity Test (1)

As a test animal for antistress activity test, a 5 week old male Sprague-Dawley rat (purchased from Biolink Co., Ltd.) was used. At 12 hours until 7 o'clock, the room temperature was adjusted to 18-23 ° C. and used in this experiment. Feed was used as a solid feed (purchased from CheilJedang Animal Co., Ltd.), and tap water was used, and feed and water supply were not limited. Three samples were used. As a method for preparing a general yeast extract, a sample obtained by adding about 10% of a Angelica extract based on autodigestion during hydrolysis at a high temperature of 50 ° C. When the hydrolyzate extract was added to about 10% based on the self-extinguishing solution after applying ultrasonic waves and vibrations, the supernatant centrifuged, that is, the supernatant of Example 2 was used as sample 2, and the precipitate precipitated after centrifugation in Example 3 Was used as sample 3, and the samples 1,2 and 3 were freeze-dried to powder and then experimented. And to see the anti-stress activity of Angelica extract only, experiments were made by making saline samples of Angelica extract 0.05, 10.0, 20.0%.

To each male Sprague-Dawley rat, each sample (1 g / kg body weight) dissolved in distilled water was orally administered once a day for 8 consecutive days. Three rats were used for each sample. Six days after the administration of the sample, the rats were placed in a cylindrical fixing frame having a width of 5 cm and a length of 12 cm, and then fixed on a ramp of 45 ° C. to induce stress for 48 hours.

3 hours after the final sample administration, rats were lightly anesthetized with ether, thymus, spleen, kidney and thyroid gland were extracted and weighed. Compared. The results are shown in Tables 2 to 4 below.

Table 2 shows the results obtained by feeding the yeast extract according to Patent Application No. 01-9946.

division Spleen (mg / 100g, B.W.) Kidney (mg / 100g, B.W.) Thymus (mg / 100g, B.W.) Thyroid gland (mg / 100g, B.W.) Normal 279.28 ± 10.24 876.62 ± 19.79 313.89 ± 21.69 6.96 ± 0.27 Stress control 216.13 ± 6.77 * 1043.83 ± 55.31 221.14 ± 21.53 5.36 ± 0.47 * Sample 1 (No Angelica) 234.11 ± 5.31 b 945.18 ± 9.95 a 236.51 ± 29.29 7.06 ± 0.59 a Sample 2 (No Angelica) 243.82 ± 16.28 a 922.26 ± 11.93 a 251.43 ± 3.83 5.71 ± 0.34 Sample 3 (No Angelica) 226.43 ± 1.84 b 1020.18 ± 44.04 211.41 ± 10.30 5.55 ± 0.69

Table 3 is the result of measuring the yeast extract according to the present invention.

division Spleen (mg / 100g, B.W.) Kidney (mg / 100g, B.W.) Thymus (mg / 100g, B.W.) Thyroid gland (mg / 100g, B.W.) Normal 288.78 ± 7.52 863.54 ± 15.33 311.25 ± 20.02 7.35 ± 0.19 Stress control 208.55 ± 8.95 * 1088.59 ± 60.82 * 219.56 ± 19.65 5.22 ± 0.25 * Sample 1 245.26 ± 4.95 a 955.32 ± 9.95 a 240.34 ± 25.65 6.50 ± 0.64 a Sample 2 266.87 ± 8.75 a 927.81 ± 12.84 a 263.11 ± 2.95 b 6.88 ± 0.47 b Sample 3 239.74 ± 2.35 b 985.30 ± 23.69 b 231.86 ± 19.65 5.96 ± 0.25

Table 4 shows the results obtained by feeding only Angelica.

division Spleen (mg / 100g, B.W.) Kidney (mg / 100g, B.W.) Thymus (mg / 100g, B.W.) Thyroid gland (mg / 100g, B.W.) Normal 263.52 ± 5.89 850.69 ± 18.23 319.68 ± 16.33 7.09 ± 0.26 Stress control 211.85 ± 3.65 * 1052.36 ± 45.62 228.37 ± 14.77 5.39 ± 0.31 * Angelica 0.5% 226.44 ± 5.81 998.65 ± 8.32 231.62 ± 18.56 5.86 ± 0.59 Angelica 10.0% 229.36 ± 5.29 987.54 ± 10.66 236.55 ± 3.29 6.02 ± 0.64 Angelica 20.0% 230.95 ± 6.12 985.33 ± 18.65 2.41.36 ± 16.22 6.13 ± 0.32

Significantly different from normal: * p <0.05, ** P <0.01

Significantly different from stress control group: a P <0.1, b P <0.05

As a result, as shown in Tables 2 to 4, when stress (stress control) is applied, organs closely related to stress hormone production and the immune system cause significant fluctuations compared to the normal group, such as the spleen, thymus, thyroid gland, etc. The weight decreased markedly and an increase in the weight of the kidney was observed. At this time, when the samples 1 and 2 were administered, it was observed that the reduction of the spleen, thymus, and thyroid gland due to stress was significantly suppressed, and it was observed that the hypertrophy of the kidney caused by stress was also suppressed. Therefore, when hydrolyzing yeast in a general way, the addition of Angelica extract and the addition of Angelica extract by ultrasound, vibration, etc., all showed some anti-stress effects. This could be attributed to the effects of hydrolysis, such as high temperature or the dissolution of raw yeast, with stresses such as ultrasound or vibration.

As a result of this experiment, the anti-stress activity of the prior patent application No. 01-9946 was stronger than the anti-stress activity, and the experiments fed only Angelica extract did not show any significant difference from the stress group. In self-extinguishing, adding Angelica extract may make antistress substances desirable.

Antistress Activity Test (2)

In addition, when stress is applied, changes in enzymes in the blood such as LDH, as well as the reduction of hypertrophy and immune cells of various immune organs such as the spleen, are known, and the increase of blood corticosteroids caused by stress affects lipid metabolism. Increases the level of cholesterol. From these findings, we measured the activity and total cholesterol of LDH, glutamate oxaloactate transaminase (GOT), glutamate pyruvate transaminase (GPT), and ALP in serum. The results are shown in Tables 5 to 7 below.

Table 5 below shows the results of the prior patent application 01-9946.

serum GOT GPT Total cholesterol LDH ALP Normal 124.18 ± 2.64 40.62 ± 1.99 81.03 ± 6.08 538.07 ± 66.39 48.97 ± 8.05 Stress control 203.6 ± 59.70 ** 58.14 ± 3.51 154.32 ± 9.73 ** 1371.07 ± 195.50 * 12.57 ± 1.29 ** Sample 1 (don't add it) 182.24 ± 21.73 43.10 ± 5.09 125.0 ± 81.51 a 718.40 ± 59.18 a 15.15 ± 1.08 Sample 2 (don't add it) 141.63 ± 3.79 a 31.50 ± 2.07 100.59 ± 1.56 b 562.40 ± 58.02 a 17.10 ± 0.90 a Sample 3 (don't add it) 212.69 ± 10.61 55.37 ± 3.37 129.04 ± 20.87 891.07 ± 21.36 19.10 ± 1.00 a

Table 6 below is the result measured after feeding the yeast extract according to the present invention.

serum GOT GPT Total cholesterol LDH ALP Normal 127.56 ± 3.41 41.98 ± 6.28 80.15 ± 5.82 543.96 ± 56.84 50.89 ± 7.52 Stress control 215.33 ± 48.22 ** 60.25 ± 8.96 152.36 ± 9.52 ** 1400.69 ± 158.36 * 13.84 ± 2.85 ** Sample 1 159.36 ± 26.35 a 40.13 ± 4.54 102.36 ± 65.21 a 607.23 ± 25.96 b 29.32 ± 2.13 Sample 2 136.59 ± 5.97 b 33.26 ± 3.85 a 90.33 ± 1.36 b 560.39 ± 39.41 b 39.01 ± 1.65 a Sample 3 179.63 ± 15.37 a 48.32 ± 4.25 110.73 ± 15.84 a 700.05 ± 19.64 a 25.33 ± 1.50

Table 7 below shows the results obtained after feeding only Angelica extract.

serum GOT GPT Total cholesterol LDH ALP Normal 123.85 ± 1.96 41.32 ± 2.00 80.69 ± 3.15 535.99 ± 54.39 49.25 ± 6.52 Stress control 207.52 ± 41.32 ** 60.49 ± 2.59 155.29 ± 8.64 ** 1402.39 ± 120.60 * 16.48 ± 3.26 ** Angelica 0.5% 190.98 ± 18.46 50.79 ± 4.23 140.69 ± 48.32 1107.35 ± 59.18 18.21 ± 1.55 Angelica 10.0% 178.63 ± 2.84 48.21 ± 7.12 131.63 ± 2.84 897.78 ± 19.65 21.36 ± 0.44 Angelica 20.0% 180.65 ± 9.69 47.92 ± 8.29 133.54 ± 14.36 921.32 ± 23.58 20.31 ± 1.88

Significantly different from normal: * P <0.05, ** P <0.01

Significantly different from stress control group: a P <0.05, b P <0.01

As shown in Tables 5 to 7, LDH and total cholesterol were increased by the stress imposition, and a significant synergistic inhibitory effect was observed due to the administration. Serum transaminase activity was increased when stress was applied and significantly decreased by administration of samples 1 and 2, especially in GOT. In addition, serum ALP activity was significantly decreased when stress was applied, and significant reduction inhibitory effect was obtained by administration of Samples 1, 2 and 3. It was found that Samples 1 and 2 had excellent antistress activity, and in particular, Sample 2 had significant antistress activity.

From the above experimental results, the anti-stress activity of the prior patent application No. 01-9946 was shown to be stronger than the anti-stress activity. The addition of Angelica extract in self-extinguishing can make desirable antistress substances.

Experimental Example 2

Anti-fatigue effect test

In order to measure the antistress activity of the yeast extract made by fermenting the present invention, anti-fatigue capacity was measured by measuring swimming endurance capacity which is closely related to antistress factors. The endurance measurement was performed using a mouse endurance measuring device (a water pool exercise device, Matsumoto et al., 1996) as shown in FIG. 1, which has high data reproducibility and reliability. As a laboratory mouse, a male mouse weighing about 28 g was 7 weeks old. (Male ICR mice: purchased at the Korea International Laboratory Animal Center) for a week, lighting time is 12 hours from 7 am to 7 pm, room temperature is 22-24oC, humidity 50% (33 × 23 × 12cm, 3 animals / room) were used for this experiment. Feed was used as solid feed (purchased from CheilJedang), and tap water was used, and feed and water were not limited. In this experiment, the control mice were orally administered a solid feed (1 g / kg), and the experimental mice were the same yeast extract and donkey extract (0.5%, 10.0) prepared in Example 1, obtained by hydrolysis at a high temperature of 50 ° C. %, 20.0%), and the yeast hydrolyzate of the prior patent application 10-2001-0009966 as a sample orally administered (1g / kg) was measured for three times of swimming time for a total of nine days. Six mice of the control group and the test group were used. To measure swimming endurance, the acrylic plastic pool (90 × 45 × 45 cm) of FIG. 4 was filled with 35 cm deep water and maintained at 34 ° C. The pump was maintained at a regulated voltage of 8 L / m with a water flow meter (type F45500, Blue white Co, Westminster, Calif., USA) and maintained at the proper conditions.

In order to minimize the deviation of the physical activity and experimental data of the mouse when measuring swimming performance, all the performance measurement time was performed from 1 pm to 5 pm, and the limit of swimming exercise time was until the mouse could not swim anymore. This was confirmed by the naked eye that the mouse does not rise to the surface of the water until 7 seconds have elapsed in the water, and set the swimming exercise time up to this point after the start of swimming. (Matsumoto et al., 1996) The results are shown in Table 8 below.

sample Swimming time by breeding period (Min) 0 days 3 days 6 days 9th Control 75 68 69 76 Yeast hydrolyzate of patent application 01-9946 74 98 116 104 Sample 1 75 105 122 120 Angelica 0.5% 76 79 86 83 Angelica 10.0% 75 78 89 84 Angelica 20.0% 74 80 79 81

As shown in Table 8, the prior patent application 01-9946 yeast hydrolyzate and the present invention Angelica fermented yeast hydrolyzed yeast extract and sugar extracts orally administered as a sample, compared to the control group prior patent application 01- It can be confirmed that the swimming time of the present invention Angelica fermentation hydrolysis is improved than the yeast hydrolyzate, and the sample of only the Angelica extract did not show a significant increase in swimming time. It was found that the anti-fatigue effect and anti-stress activity effect on the extract was superior to the yeast hydrolyzate of prior patent application No. 01-9946, and the effect of only Angelica extract was found to be inadequate.

The present invention reveals a novel use of yeast extract and Angelica extract as an antistress and neurotrophic agent, which is used only as a normal food raw material, and thus can be utilized as a medicine and a functional food as an alternative to a conventional antipsychotic agent having many side effects.

In addition, natural neuronutrients can be easily prepared by adding yeast peptides, which are natural herbs when hydrolyzing food grade yeast, instead of neurotrophic factor (neutropin), which has been hardly obtained through conventional recombinant strain expression. In addition, it was manufactured to cross the cerebrovascular barrier by oral administration. As a new use of yeast extract and donkey extract, anti-stress effect and anti-fatigue effect were developed. Thus, yeast extract and donkey extract are used as functional foods. In addition, it can be used as a raw material for pharmaceuticals.

Claims (12)

Culturing the yeast strain to the maximum growth phase; And Method of producing a yeast hydrolyzate comprising a fermented Angelica extract, comprising: fermented Angelica extract with anti-stress, anti-fatigue function comprising the step of adding the donkey extract to the cultured yeast at a temperature of 35 ℃ to 70 ℃. The method according to claim 1, further comprising the step of performing hydrolysis by adding proteolytic enzymes simultaneously with or after autodigestion, wherein the fermented Angelica extract has antistress and anti-fatigue functions. Process for producing yeast hydrolyzate comprising. [Claim 2] The yeast comprising the fermented Angelica extract, which has the anti-stress function and the anti-fatigue function, further comprising the step of centrifuging the digested yeast hydrolyzate to separate only the supernatant. Process for producing hydrolyzate. The method according to claim 3, wherein the supernatant taken by centrifugation is filtered using an ultrafiltration ultrafiltration membrane to take only a peptide having a molecular weight of 10,000 or less. Process for producing hydrolyzate. The fermented Angelica extract according to claim 1, wherein the Angelica extract is obtained by heating and extracting Angelica with ethanol at 40 to 100 ° C, followed by drying under reduced pressure. Manufacturing method of yeast hydrolyzate containing. 6. The method of claim 5, wherein the Angelica extract is added in an amount of about 0.1 to 30% by weight, wherein the yeast hydrolyzate comprising fermented Angelica extract has antistress function and anti fatigue function. The method according to claim 1, further comprising the step of improving the yeast strain by selecting only the surviving strains by culturing with application of high temperature, ultrasonic wave, vibration, pH change, etc. in the range in which yeast is not killed before culturing the yeast to the maximum growth stage. A method for producing a yeast hydrolyzate comprising fermented Angelica extract, which has an antistress function and an antifatigue function. The method of claim 1, further comprising inducing the secretion of metabolites to withstand stress by applying appropriate physical or chemical stresses such as high temperature, ultrasonic waves, vibrations, pH changes, etc., in the range that yeast does not die before self-extinguishing the yeast. A method for producing a yeast hydrolyzate comprising the fermented Angelica extract, which has an anti-stress function and an anti-fatigue function. An antistress preparation comprising the yeast hydrolyzate comprising the fermented Angelica extract, prepared according to any one of claims 1 to 7, as an active ingredient. The agent according to claim 8, wherein the antistress agent has an activity of nerve stabilization and tension relaxation. A fatigue recovery preparation comprising the yeast hydrolyzate comprising the fermented Angelica extract, prepared according to any one of claims 1 to 7, as an active ingredient. A neurotrophic agent prepared according to any one of claims 1 to 7, comprising a yeast hydrolyzate comprising the fermented Angelica extract as an active ingredient.