KR100416210B1 - Peptide derived from yeast having activities as anti-stress, anti-fatigue and natural brain-neurotrophic factor and preparing process thereof - Google Patents

Abstract

PURPOSE: A functional peptide obtained by hydrolysis of yeast having an antistress and antifatigue function and a function as a neurotrophic factor is provided, which can be continuously taken everyday without causing any problem in safety and can relieve and prevent mental and physical symptoms caused by stress and fatigue. CONSTITUTION: This functional peptide is obtained by self-digesting yeast at 35 to 70deg.C, hydrolyzing protein with protease, centrifuging, filtering a supernatant using an ultrafiltration membrane and separating a yeast peptide having a molecular weight of less than 10,000. The yeast peptide contains additionally 15 to 25% by mole of glutamic acid and aspartic acid respectively.

Description

Peptide derived from yeast having activities as anti-stress, anti-fatigue and natural brain-neurotrophic factor and preparing process according to anti-stress, anti-fatigue and brain management factors

The present invention relates to a yeast-derived functional peptide having an antistress function, an anti-fatigue function and a natural brain renal quantitative factor, and a method for producing the same, and more specifically, to hydrolyze a yeast licensed as a protein-based health functional food material. And a functional peptide having an antistress function, an antifatigue function and a function as a brain neurotrophic agent obtained by purification.

Modern people live under various stresses, and the various emotional states caused by these stresses affect the health by causing changes in the autonomic nervous system, hormones and immune function in the body.

In general, under stress, hormones are secreted by the excitability of the sympathetic nervous system in the autonomic nervous system.In particular, the secretion of adrenaline is promoted (Corrodi H, Fuxe K, Hokfelt T. The effect of immobilization stress on the activity of central monoaminergic neuron, Life Sci 7: 108-112, 1968). The human body tries to maintain the balance of the entire autonomic nervous system by stimulating the parasympathetic nerve to secrete acetylcholine in order to prevent this imbalance, that is, to prevent this continuous sympathetic nerve excitement. However, if you continue to receive stress, your body becomes exhausted and can no longer effectively manage the stress energy accumulated in the sympathetic nervous system, which eventually leads to a breakdown of the autonomic nervous system and directly affects cellular and humoral immune mechanisms, resulting in a decrease in immune function. If a disease occurs and this condition persists, an organic disease occurs that eventually results in an organic disorder.

This causes many symptoms of illness, such as peptic ulcer, hypertension, cancer, diabetes, irritable colitis, heart disease, bronchial asthma, tension headache, arthritis, neurodermatitis. In addition, the symptoms of the whole body feel drowsy and tired easily, there is no endurance in life, you can't sleep deeply, you sweat a lot, you feel cold, you feel pain in your shoulders, your chest is stuffy, and you feel something hanging around your neck. , Dizziness, decreased libido in men and infertility in women.

Conventionally, diazepam, meprobamate, methylpentinol, and ethoxyxin as an anxiolytic agents to improve the symptoms of such stressful diseases; Chloropromazine, promethazine, and azaferon as neuroleptics; Bunitrol as a beta-adrenergic blocker; Tricyclic, tetracyclic compounds used alone or in combination with neuroleptics as antidepressants; Caffeine, amphetamine or derivatives thereof as psychostimulant; And it is known to use psychotropic drugs such as combinations of phenobarbital and codeine as sedatives and hypnotics (Poldinger, W., Schmidlin, PE, Wider, F., Index Psychopharmacorum, H. Huber, Bern). Drugs have the disadvantage of showing only predominant symptomatic drug treatment without causative pathology to stress, and these agents do not reduce chronic damage caused by stress, do not counteract stress through catabolism regeneration, It modifies the normal (antistress) response of body adaptation and causes a number of adverse reactions. In addition, anxiolytics and psychostimulants (with amphetamine and caffeine types) often need to be increased due to the effects of acquired resistance, and serious problems exist that make them dependent on psychoactive substances as an important adverse reaction.

Therefore, in order to solve the stress, that is, the imbalance of the autonomic nervous system, the solution of different angles is being sought, and the contents are as follows. In other words, stress is a medically harmful stimulus that breaks the body's equilibrium, and neurotransmitters are involved in the body's response to this stimulus. There is acetylcholine derived. Acetylcholine is a neurotransmitter that promotes relaxation and occurs in the parasympathetic nerve, the autonomic nervous system. In addition, there are about 50 neurotransmitters discovered so far.

These neurotransmitters must have a complementary target called a receptor to function properly. Even if there are many neurotransmitters, they cannot function without the receptor of the peptide structure that is complementarily bound to the neurotransmitter. none. Muscarinic receptors that bind to acetylcholine consist of peptides comprising aspartic acid and glutamic acid residues that play an important role in binding and the group of hydrophobic amino acids surrounding them (Gearien 1999). Therefore, when a neurotrophic factor containing such peptides is supplied to the body, nutrition is supplied to the nerve cells to induce the growth of the nerve cells and to solve various neurological diseases such as Parkinson's disease without side effects and sufficient nerves. Through promoting the production of receptors of the transport material and neurotransmitter, it is possible to solve the disease caused by stress by preventing the overload by absorbing and transmitting the nerve impulse (impulse) due to excessive stress (stimulation) well. Therefore, it is also important to consume a lot of neurotransmitters to strengthen the nervous system, but it is also important to consume a lot of peptides, ie neurotrophic factors that help the receptor production of neurotransmitters. In addition, even if the peptide acts as a receptor, the reaction proceeds according to the amount of the neurotransmitter body, but no further reaction proceeds, and thus side effects due to overdose are not generated.

Herein, the brain-neurotrophic factor refers to a factor that acts as a nutrient for nerve tissues such as the brain and spinal cord to help the growth of nerve cells or glial cells. In the meantime, it has been found that brain neurons do not grow, but the fact that brain neurons maintain growth and differentiation by supplying specific nutrients to the brain has emphasized the importance of neurotrophic substances, thereby nourishing neurons. Neurotropins, which are supplied to facilitate the maintenance and regulation of neuronal growth and differentiation, were searched and extracted from the brains of fetuses and animals to reveal their peptide sequences. As such, neurotrophic factors consisting of peptides, which stimulate the growth or proliferation of nerve tissues to promote the survival and differentiation of nerves or glial cells, are an important factor that can solve neurological diseases and occur in neurodegenerative and neurodegenerative diseases. It has been shown to be useful for the treatment of dysfunctional symptoms such as Parkinson's disease or Alzheimer's disease (Varon and Bunge 1997. Ann. Rev. Neuroscience 1: 327: Thoenen and Edgar 1985 Science 229: 238).

Important neurotrophic or neurotrophic factors discovered so far include nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and glial cell-derived neurotrophic factor (GDNF) derived neurotrophic factor), neurotrophin-3 (NT-3) and neurotropin-4 / 5 (NT-4 / 5), which are also proteins and are composed of peptides. However, in order to search for and discover new neurotrophic factors, it is necessary to use human brain or fetal brain. Since the amount of the nutrients is very small, it is difficult to detect and should be removed after death. Is just a few. Rather, it is estimated that there are many more types of neurotrophs than those found so far.

In addition, until now, neurotrophic factors or neurotransmitters have been artificially manufactured in the form of synthetic peptides or recombinant peptides using a peptide synthesizer or a gene recombination method through PCR amplification. It is not easy to use, and most of them cannot reach the brain neurons by oral administration because they cannot cross the Brain Blood Barrier with macromolecules of 30,000 Daltons or more. There are drawbacks that must be made (Medical Report 1998, January and February).

Therefore, the yeast extract or yeast peptides are prepared from food grade yeast as described in the present invention without using complicated processes such as genetic recombination methods, and thus act on the nervous system in the human body even in a relatively small dose to prevent anti-stress, anti-anxiety and sleep. If it exhibits neurophysiological activity such as induction, and anti-fatigue effect through stress suppression, this is compared with the existing neurotrophic factors (neutropin, NT-3, BDNF, NGF, etc.) discovered by genetic recombination. It also has excellent commercial utility and can address safety issues that may be suspected of being genetically modified (GMO).

Yeast is recognized as GRAS grade which is harmless to the human body, and more than 50% contains high quality protein, a large amount of minerals and vitamin B group, so it is useful for liquor and bakery industry as well as various useful substances such as proteins, nucleic acids, enzymes, lipids. , Vitamins, minerals, etc. (Roman et al., Food Biotechnology, 6, 225, 1992) Yeast extract produced by hydrolysis with yeast autolysis enzymes and other proteolytic enzymes ) Has been used as a raw material for microbial fermentation media, seasonings and health foods (Bioindustry, 14, 53, 1997). However, the yeast extract or yeast-derived peptides produced by hydrolysis of yeast have not yet been used for specific purposes as anti-stress and brain neurotrophic agents (natural neurotrophic factors), and there have been no examples of experiment confirmation and analysis. It has not been used as an antistress agent, anti-anxiety agent, or sleep aid, nor has it been published.

The present inventors anaerobic or aerobic reaction according to the environment of the growth medium and stop the growth when the environment is not good, and cope with the external poor environment through heterozygosity, etc. It has a function that responds sensitively to the effects of the first time. All organisms have a warning response to stress when they first perceive stress, and a physiological arousal reaction that mobilizes body resources occurs. In response to stress, the efforts to cope with stress are effective, and the physiological changes are stabilized (Seyle, 1956). After growing the yeast to the maximum growth period, it is possible to perform normal growth activities within the extent that yeast is not killed. Without adequate physical or chemical stress in the absence of high temperature, ultrasonic or vibration, pH Yeast extract is added to the growth medium to produce and secrete a large amount of anti-stress substances (proteins, enzymes), and then yeast extract made by hydrolyzing yeast by autodigestion or proteolytic enzymes, and purified by ultrafiltration membranes. The present invention has been newly discovered that the derived peptide has a function as an agent for treating and preventing autonomic neurological diseases such as an antistress agent, an anxiety agent, a sleep aid, and a brain neuronutrient (natural neurotrophic factor).

Therefore, an object of the present invention is to provide a new yeast extract and functional peptide (peptide) having the function of anti-stress, anti-fatigue and natural brain renal quantitative factor obtained by hydrolysis of yeast.

In addition, the present invention aims to provide a complex natural neurotrophic factor comprising the yeast extract or yeast peptide as an active ingredient in order to solve the disease of the human body caused by stress and maintain the balance of the autonomic nervous system.

In addition, in the present invention, for the treatment of neurological diseases, including autonomic nervous system diseases caused by stress, the anti-stress, anti-anxiety, nerve stabilizing, anti-stable activity comprising the yeast extract or yeast peptide as an active ingredient We aim at providing stress agent

In addition, an object of the present invention is to provide a fatigue recovery agent comprising a yeast extract or a yeast peptide as an active ingredient having an anti-fatigue effect on exercise muscle fatigue, which is closely related to stress.

In addition, the present invention is to provide a functional drink containing a natural anti-stress material extracted from a natural-derived material that is proven to be safe and not a drug having serious side effects, so that it can be easily and safely ingested stress relief material There is.

1 is a diagram schematically showing a muscarinic acetylcholine receptor map,

2 is a graph showing the results of the anti-stress experiment (1) of the present invention,

3 is a graph showing the results of the anti-stress experiment (2) of the present invention,

4 is a view showing the endurance measuring device for the mouse (measuring pool exercise device) for measuring the anti-fatigue effect,

5 is a graph showing the results of the anti-fatigue test of the present invention,

Figure 6 is a photograph showing the results of clinical trials of the anti-stress effect on the human body of the present invention.

In order to achieve the above object, the present invention provides a yeast extract having a function as an anti-stress function, an anti-fatigue function and a neurotrophic factor, which is extracted from yeast.

In the yeast extract of the present invention, the yeast contains a large amount of high quality protein, minerals and vitamin B group, and the yeast is extracted using a conventional extraction method known in the art, such as autolytic enzyme or protease. can do. In addition, it is found that the existing yeast extracts on the market function as antistress agents, anti-fatigue agents and brain management medications according to the present invention.

In addition, the present invention provides a yeast extract having a function of anti-stress function, anti-fatigue function and neurotrophic factor, characterized in that the yeast is auto-digested at a temperature of 35 ℃ to 70 ℃, preferably 50 ℃ to 60 ℃. .

Yeast extract of the present invention can provide a yeast extract having an effective anti-stress activity only by self-extinguishing at a high temperature of 35 ℃ or more, which obtains the effect of stressing yeast bacteria by applying high temperature, It is thought to be due to the release of stress-resistant substances to resist stress at the same time as self-extinguishing. Furthermore, by selecting stressed strains that are resistant to stress by applying stress such as ultrasonic waves or vibrations, and applying the stresses to the strains again to produce more stress-resistant substances, an extract having better antistress activity is produced. can do.

In addition, the present invention is characterized in that the yeast extract having a function as an anti-stress function, anti-fatigue function and neurotrophic factor, characterized in that the protein is hydrolyzed by the protease after simultaneously or auto-digesting the yeast as described above To provide.

In addition, the present invention provides a yeast extract having a function as an anti-stress function, an anti-fatigue function and a neurotrophic factor, characterized in that only the supernatant is taken by centrifuging the hydrolyzed yeast as described above.

In addition, the present invention provides a yeast peptide having a function as an anti-stress function, an anti-fatigue function and a neurotrophic factor, characterized in that the supernatant centrifuged as described above was filtered using an ultrafiltration ultrafiltration membrane to obtain a molecular weight of 10,000 or less. to provide.

In the yeast peptide of the present invention, the molecular weight cut-off to 10,000 is generally referred to as a peptide having a molecular weight of 10,000 or less because the peptide is involved in a bioregulatory function showing various physiological activities as a functional food material (2000 new technology Trend Research Report, Functional Foods, Patent Office p41), especially for the protection of cerebrovascular vessels, is intended to facilitate the passage of the brain blood barrier in the human body, which prevents the passage of cerebrovascular vessels. .

The present invention also provides a yeast peptide having a function of anti-stress function, anti-fatigue function and neurotrophic factor, characterized in that 15 to 25 mol% of glutamic acid and aspartic acid are contained.

Yeast peptides of the present invention are rich in glutamic acid and aspartic acid, which are amino acids capable of forming a muscarinic receptor to which acetylcholine binds, and thus have a great effect on stress diseases caused by imbalance of the autonomic nervous system.

In addition, the present invention comprises the steps of culturing the yeast strain up to the growth stage; Hydrolyzing the cultured yeast at a temperature of 35 ° C. to 70 ° C. while simultaneously adding more proteolytic enzymes to hydrolyze the yeast protein; And it provides a method for producing a yeast extract having a function as an anti-stress function, anti-fatigue function and neurotrophic factor comprising the step of separating and separating only the supernatant by centrifuging the hydrolyzed.

In the method for preparing the yeast extract of the present invention, the yeast strain is preferably selected by culturing only the surviving strain by applying high temperature, ultrasonic wave, vibration, pH change, etc. in the range that yeast is not killed before culturing the yeast to the maximum growth stage. It further comprises the step of improving.

In addition, the production method of the yeast extract of the present invention, metabolism to withstand stress by applying a suitable physical or chemical stress such as high temperature, ultrasonic wave, vibration, pH change in the range that yeast does not die before the yeast self-extinguishing It further comprises the step of inducing secretion of the substance.

This is because when all organisms perceive stress for the first time, a warning response is generated against stimulation, and a physiological arousal reaction that mobilizes physical resources occurs.When stress continues, the organism enters a resistance phase and its efforts to cope with stress are effective. On the basis of its stabilization (Seyle, 1956), when a yeast is also stressed, it secretes various enzymes to counter the stress. The secreted enzyme is a low-molecular-weight protein, which is a neurobiological agent that is beneficial to the stressed human body. The idea is to work.

In the production method of the yeast extract of the present invention, the physical or chemical stress applied to the yeast to induce the production and secretion of the anti-stress substances of the yeast, 35 ℃ to 45 ℃ temperature higher than the optimum growth temperature within the range that yeast is not killed Stimulation such as high temperature, ultrasonic wave or vibration, and pH change.

Method for producing a yeast extract of the present invention, more preferably strain improvement step of selecting only the surviving strain cultured by applying ultrasonic and vibration to the yeast strain; Inoculating the selected strains on YM medium and incubating the cells at a logarithmic growth phase at 22 ° C. to 25 ° C., followed by centrifugation, diluting the cells with 1% pepton buffer; Applying a high temperature of 35 ° C. to 45 ° C. and a stress of ultrasonic wave and vibration for 8 hours that is higher than the growth optimum temperature so that the yeast generates and secretes metabolites to withstand the stress; Hydrolyzing the yeast protein by further adding proteolytic enzymes while auto-digesting at a temperature of 50 ° C. to 60 ° C., and separating the supernatant by centrifuging the hydrolyzed ones. do.

In addition, the present invention is characterized in that the anti-stress function, anti-fatigue, comprising the step of filtering only the supernatant taken by centrifugation in the method of preparing the yeast extract using ultrafiltration ultrafiltration membrane to take only a peptide having a molecular weight of 10,000 or less. It provides a method for producing a yeast peptide having a function and a function as a neurotrophic factor.

In the production method of the yeast peptide of the present invention, the supernatant taken by centrifugation is filtered using an ultrafiltration membrane for cut-off 10,000 ultrafiltration to take only a low molecular weight peptide of 10,000 or less to reduce the efficacy of the extract. It removes and facilitates the absorption of the human body, so that only low-molecules with high bioavailability can be separated and purified, and in particular, it can easily pass through the blood-brain barrier.

In the yeast extract or yeast peptide production method of the present invention, it may further comprise the step of drying the yeast extract or yeast peptides prepared by the above method to a powder state.

In addition, the present invention provides an anti-stress preparation comprising the yeast extract or yeast peptide prepared by the above-described preparation method as an active ingredient.

The antistress agent of the present invention is characterized by having an activity such as nerve stabilization, tension relaxation, anxiety removal or sleep assistance.

The present invention also provides an anti-fatigue agent (fatigue recovery agent) comprising a yeast extract or yeast peptide prepared by the above-described preparation method as an active ingredient.

The present invention also provides a neurotrophic agent (new management quantitative factor) comprising the yeast extract or yeast peptide prepared by the above-mentioned manufacturing method as an active ingredient.

In addition, the present invention provides an anti-stress, anti-fatigue and neurotrophic functional beverage comprising the yeast extract or yeast peptide prepared by the above-described preparation method.

The functional beverage of the present invention stresses the proven yeast bacteria to secrete anti-stress substances, and by preparing a beverage from yeast extracts or yeast peptides made to contain such anti-stress substances, it is safe and easy to take, but also neurostable It is possible to provide an anti-stress beverage having an effect such as relaxation, tension and sleep support.

The functional beverage of the present invention is preferably characterized in that it comprises 0.1 to 10% by weight of yeast extract or yeast peptide, 10 to 25% by weight of ordinary beverage additives such as sweeteners and acidulants, and the remaining weight% of water. Typical beverage additives in the functional beverage of the present invention may include liquid fructose, sugar, maltodextrin, glucose, citric acid, nicotinic acid amide, pantothenic acid, sodium benzoate, flavoring agent, flavoring agent, and the like. Further juices may be added to prepare the beverage.

In addition, the present invention comprises the steps of culturing the yeast strain up to the growth stage; Hydrolyzing the cultured yeast at a temperature of 35 ° C. to 70 ° C. while simultaneously adding more proteolytic enzymes to hydrolyze the yeast protein; Centrifuging the hydrolyzed one to separate and separate only the supernatant; Putting the supernatant into water with activated carbon, stirring and sterilizing under pressure; And it provides a method for producing an anti-stress, anti-fatigue and neurotrophic functional beverage comprising the step of purifying the sterilized under reduced pressure to colorless.

In order to make the yeast extract containing the antistress material into a beverage preparation, the yeast-specific odor must be removed by a purification process. As a purification method for this purpose, the prepared yeast extract is mixed with water and activated carbon, stirred, and filtered by autoclaving. This is mixed with activated carbon for the purpose of decolorization and deodorization by using the property that amino acid is hardly adsorbed to active carbon (Song Hyung-ik, new mesenchymal, modern fermentation engineering, 1998, 293).

The filtered beverage concentrate is diluted to produce an antistress beverage. The beverage is prepared according to the conventional method for preparing a beverage, and is not particularly limited as long as it includes a yeast extract prepared according to the method of the present invention.

The method for producing a functional beverage of the present invention preferably comprises the steps of improving the yeast strain by selecting only the surviving strains by culturing while applying high temperature, ultrasonic wave, vibration, pH change, etc., in a range that does not kill yeast, and yeast is killed. It may further comprise the step of inducing the secretion of metabolites to withstand the stress by applying a suitable physical or chemical stress such as high temperature, ultrasonic wave, vibration, pH change in the range that does not.

In the production method of the functional beverage of the present invention, the self-extinguishing temperature is preferably 50 ° C. to 60 ° C., and autoclaving is preferably performed at a pressure of about 1.5 at a pressure of about 1.5 and at a temperature of 100 ° C. to 125 ° C. for 10 to 15 minutes.

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

In order to normalize the changes in the various organs caused by the imbalance of the autonomic nervous system, that is, the continuous sympathetic nerve excitement due to stress, the excitatory transmission of the parasympathetic nerve should be smoothed so that the change on each organ is normal. To this end, acetylcholine, a neurotransmitter at synapses, must be secreted well, and acetylcholine receptors must be present in order for acetylcholine to be transferred from the synapse to the next nerve cell. Acetylcholine binds to muscarinic receptors as shown in FIG. Therefore, in order for acetylcholine to be well-transmitted at synapses, there must be many muscarinic receptors at synapses, and many muscarinic receptors must be made. The muscarinic receptor is composed of seven segmented proteins, and the quaternary ammonium salt of acetylcholine is important for muscarinic activity in order to act as a double receptor. It is thought to be due to acidic amino acid residues. Possible aspartic acids are aspartic acid and glutamic acid (see Kang Kun-il, 1992, Introduction to Medicinal Chemistry 10: 131).

Therefore, among the peptide preparations that can be used as a raw material for forming the muscarinic receptors of acetylcholine, the peptide preparations having an amino acid composition rich in the acidic amino acid content of aspartic acid and glutamic acid facilitate the synthesis of muscarinic receptors in the human body. By influencing the efficient delivery of acetylcholine in the nerves, it is possible to fundamentally prevent and cure stress and mental illnesses by balancing the autonomic nervous system imbalance with the constant excitability of the sympathetic nerves caused by stress.

The present inventors have found that the yeast extract is rich in amino acids capable of forming muscarinic receptors as described above, and thus, it can be concluded that the yeast extract has a great effect on stress diseases caused by the imbalance of the nervous system. there was.

The yeast extract of the present invention contains 18 different amino acids as shown in Example 4 below, and particularly, aspartic acid and glutamic acid have a very high content. Both of these amino acids are amino acids with a negatively charged R group. These results include a lot of negatively charged amino acids that are essential for the muscarinic receptor, a neurotransmitter of acetylcholine, a neurotransmitter in the parasympathetic nervous system, which can be used as a good raw material when acetylcholine receptors are synthesized in the body. Indicates that it can. Thus, the yeast extract of the present invention may have antistress function by helping to smooth the neurotransmission of parasympathetic nerves to balance the sympathetic nerves that are excited during stress.

Food grade yeast is recognized as GRAS grade which is harmless to human body and contains more than 50% of high-quality protein, a large amount of minerals and vitamin B group, so that the peptide which can be a raw material of neurotransmitter or its receptors Not only is it abundant, but in particular, the B vitamins and minerals in yeast act not only for normal energy metabolism, but also for activating the brain and nerve cells, so that vitamin B ₆ (pyridoxin) is used for serotonin, dopamine, norepinephrine, GABA, and taurine. It is an indispensable nutrient for synthesizing the same neurotransmitter.If it is insufficient, it is anxious, unfocused, memory is low, and vitamin B ₁ (thiamin) deficiency causes memory problems such as memory disorder, numbness, dementia, and B2 is the brain. It can act as a coenzyme necessary for growth of neurologically important neurotrophic factor.

Yeast is also a good source of selenium, which is an essential source of antioxidant enzymes in the production of antioxidant enzymes, which are important for brain nerve cell growth. Yeast is also a good source of selenium. And become anxious. Yeast, in particular, is a form of mineral enriched yeast that forms organic chelating bonds with minerals such as chromium, including selenium, and also acts as a supplementary source of organic minerals that are vulnerable to human consumption (US Patent 4.530.846). No. 1985.7.23 Selenium yeast production method) in view of this fact. Yeast acts as an excellent brain neurotrophic substance in the form of yeast extracts or yeast peptides, and is a functional food material and natural peptide material that has excellent effects on anti-stress, anti-anxiety, nerve stability, relieving sleep disorders caused by stress, and fatigue recovery. It can be said.

In addition, since the various secretory substances (exoenzyme) produced in the present invention are secreted through the cell wall of the yeast is to be produced a large amount of low molecular weight protein so that the cell wall of the yeast acts as a natural membrane filter to pass through the cell wall Eating it can easily cross the blood-brain barrier in the human body, which can act as a brain neurotrophic substance.

In addition, the yeast has a self-extinguishing function, and when the yeast is in an environment where it is far out of conditions (temperature, pH, etc.) or hunger without nutrients, the cytoplasm of the yeast is hydrolyzed by the action of its own enzyme. In this case, the cell wall is also destroyed by continuous enzymatic reaction, and the already decomposed various cytoplasm is eluted out of the cell through the broken cell wall to increase the bioavailability of the yeast. Through the production of large amounts of low-molecular weight antistress substances, the self-digestion of the yeast dissolves various proteins, minerals, and vitamin B groups contained in the yeast and acts as a more efficient neurotrophic substance, thus providing anti-stress effects. Eggplants can obtain yeast peptides.

In the production method of the yeast extract of the present invention, by adding a step to induce the yeast to produce a material against it by applying stress such as high temperature, ultrasonic waves and vibration to the yeast is to kill the useful components of the yeast cell Yeast loses the selective permeability of cell membrane by applying higher temperature than temperature, and various kinds of taste substances (glutamic acid, etc.) through self-extinguishing step that the cell membrane is lost by using enzymes such as protease, lipase, invertase, maltase, zymase etc. Amino acid, 5′-AMP, and other nucleic acid degradation products, etc.) (see Song Hyung-ik and Shin, Shin-Yeop, Hyundai Fermentation Engineering, 1998. 5: 189), and at the same time, the addition of papain, a proteolytic enzyme, for more effective antistress activity, To obtain anti-fatigue activity and neurotrophic agents. Generally, temperature affects the growth and survival of yeast. Note is one of the most important parameters. Most microorganisms are mesophilic, and yeast has a growth temperature limit of 20-46 ° C. A mild heat shock of 37 ° C for Saccharomyces cerevisiae, a mesophilic yeast, is 48- It also induces resistance to 55 ℃, and these heat-resistant cells induce heat shock even when the temperature rises only 5 ℃ above the optimum growth temperature (35 ℃), and has a molecular weight of 104, 90, 70, 60kDa with trehalose per peat. Essential heat shock proteins are synthesized. (See Michelle L. Deegenaars and Kenneth Watson, 1998. Environmental Microbiology. August.) This is a good example of how yeast synthesizes the antistress proteins needed to fight heat stress. With this in mind, the present invention adds a manufacturing step to induce the yeast to produce a material that counters the stress by applying high temperature, ultrasonic and vibration stress to the yeast, while the yeast bacteria auto-digest at a high temperature of 35 ℃ to 70 ℃, The yeast extract and yeast peptides were prepared, further comprising the step of hydrolyzing the yeast protein by further adding a degrading enzyme, and centrifuging the hydrolyzed one to separate the supernatant.

In order to test whether the yeast extract and yeast peptide of the present invention has an anti-stress capacity through the neurotrophic function, the test method is as in the following experiments Breckman 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.) Using these factors in mind, immobilized stressed rats were used to determine the changes in organ weight and blood biochemical factors affected by stress. The change was measured and found to have an antistress effect.

In addition, as an anti-stress test for the yeast extract and yeast peptide of the present invention, an anti-fatigue effect test was performed by measuring 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, the yeast extract and yeast peptide of the present invention were confirmed to have excellent anti-stress effect and anti-fatigue effect.

In addition, since the yeast extract and yeast peptides according to the present invention have an antistress function, in general, when the stimulus affecting the living body becomes a certain intensity or more, it is injured, irrespective of the type of stimulus, tension headache, migraine, high blood pressure , Indigestion, fatigue, headaches, and if chronically caused nonspecific systemic adaptation syndrome (general adaptative syndrome) such as causing various neurosis, gastric damage (Hwang, Wan-wan, 心身 症, Seoul, Haenglim Publishing Co., pp. 17-28, 33-49, pp. 277, 1985) Through the suppression of stress, the yeast extract and yeast peptide of the present invention can be used for the use of neuro stabilizers, relaxants, etc. It is highly expected to be used as a sleeping aid for those who cannot sleep. In addition, it can be usefully used as a dietary supplement and special nutritional food as a functional food material having a function as a brain and neurotrophic substance containing such a use.

Therefore, in light of this, yeast peptides and yeast extracts seem to play a role in autonomic nerve regulation, such as to relax tension and induce sleep, and the present invention yeast peptides and yeast extracts in normalizing nerve function, It is a very effective neuronutrient that has little side effects even when ingested in large amounts.

Solve various stress diseases through yeast extract and yeast peptides that have anti-stress effects and functions as natural neurotrophic agents without the side effects of conventional neuropathy and stress disorder treatments (neural stabilizers, anti-anxiety agents, sleeping pills, etc.) The recovery of autonomic nervous system function can be used as a nerve stabilizer, anti-anxiety, sleep aid, and fatigue recovery.

Yeast extract and yeast peptides as an antistress and neurotrophic supply function of the present invention can be used in a variety of medicines, animal drugs, functional foods, etc. as a substitute for a lot of side effects psychotropic agents. In order to be used as a therapeutic agent in this way it can be prepared by a known method in the pharmaceutical field, and mixed with itself or a pharmaceutically acceptable carrier, excipient, diluent or the like powder, granules, tablets, capsules, or injections It can be prepared and used in the formulation of. They can also be administered orally or parenterally.

Dosage to administer the yeast extract and yeast peptides according to the present invention as an active ingredient may be appropriately selected according to the age, sex, condition, symptoms of the disease of the patient, preferably 500mg per day, adult insomnia In the case of honey, 250mg powder capsule is administered twice a day.

Example

Although the present invention will be described in more detail with reference to the following examples, the examples should not be construed as limiting the present invention, and conventional variations of those skilled in the art are possible within the scope of the present invention.

Example 1

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

The culture broth cultured as above was centrifuged at 15,000 rpm for 15 minutes to remove the supernatant, and the remaining precipitate, that is, 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, 1% of protease (papaine 30.000) was added to facilitate the breakdown of yeast proteins into peptides. Time hydrolysis was performed to dissolve the protein into peptides.

Thus dissolved was centrifuged at 15,000 rpm to separate only the supernatant in the aqueous solution to obtain the yeast extract of the present invention. The thus obtained was lyophilized to be referred to as "sample 1" and used in the following experimental example.

Example 2

1) Selection of improved strains of yeast

Food-grade Saccharomyces cerevisiae 7904 strains from KCTC were incubated for 48 hours under vibrational stress conditions using a 45 kHz continuous ultrasound and a dropping width through an underwater pump. Surviving yeast was screened on YM agar plates (3 g yeast extract, 3 g malt extract, 5 g peptone, 10 g glucose, 15 g / L agar) to select the fastest growing and large colonies on the plate as improved strains.

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, in order to promote the hydrolysis of the yeast protein into peptides, 1% of a protease (papaine 30.000) was added and hydrolysis was performed for 48 hours at an initial pH of 4.0 to dissolve the protein into peptides.

The percent hydrolysis 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 then diluted 500 µl of the diluted solution into 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 55%.

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 in an aqueous state, which was used in the following Experimental Example as "Sample 2," and the precipitate as "Sample 3". Also used in the following experimental example. The supernatant and the precipitate thus obtained were lyophilized to form a powdered 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 yeast extract obtained in Example 2 was dehydrated with cold acetone, dried on a filter paper in a 60 ° C. dry oven, dried, and 5 mg of a sample was added to a hard test tube, 5 ml of 6 N HCl was added, and then degassed and sealed at 110 ° C. Hydrolyzed for 24 hours in a small amount of water and washed with a small amount of distilled water, and then dried at 50 ° C. in a concentrator to remove hydrochloric acid, and dissolved buffer to perform column Li 10 cm No. 338051 ion exchange with an automatic amino acid analyzer (Deckman system 6300, USA). 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 14.2 Leucine 6.9 Lysine 5.2 Aspartic acid 5.0 Alanine 4.7 Histidine 4.3 Isoleucine 3.9 Phenylalanine 3.7 Valine 3.4 Tyrosine 3.5 Threonine 3.0 Glycine 3.0 Serine 2.8 Arginine 2.3 Proline 2.0 Tryptophan 1.4 Methionine 1.5 Cysteine 0.8

As shown in Table 1, the yeast peptide prepared in the present invention contains 18 different amino acids, and particularly, the content of aspartic acid and glutamic acid is very high.

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.

Example 5

Manufacture of beverages

1) Filtration and Purification

The supernatant obtained in Example 2 was mixed with the same amount of activated carbon (5% of the weight of the supernatant), and then stirred in an amount of water of about 20 times. This stirred product was put in a autoclave sterilizer for 15 minutes at 121 ℃ and filtered under reduced pressure twice. Decolorization Deodorized colorless purified yeast extract was obtained.

2) Preparation of Beverage Composition

The yeast extract prepared in 1) was diluted in 70 parts by weight of water, and 7 parts by weight of liquid fructose, 4 parts by weight of glucose, and 5.9 parts by weight of plum juice concentrate were added thereto to prepare a beverage composition.

3) additive effect

When manufacturing anti-stress beverages from yeast extracts, the taste of raw materials is low, so sweetening and acidifying agents have been added to improve their palatability. No, 3: normal, 5: very good), the sensory test was carried out and the results are shown in Table 2 below.

Subsidy color incense flavor General preference Remarks Sweetener Liquid fructose 3% 4.2 3.8 2.5 3.5 ※ 5% of plum juice concentrate 5% of liquid fructose 4.2 3.9 3.0 3.7 Liquid Fructose 7% 4.4 4.1 3.5 4.0 Liquid Fructose 9% 4.2 4.1 3.5 3.9 Glucose 2% 4.0 3.9 2.5 3.5 Glucose 4% 4.0 3.9 3.5 3.8 Glucose 6% 4.0 3.9 3.2 3.7 Acid Plum Juice Concentrate 4% 3.5 3.4 3.9 3.6 ※ 8% liquid fructose combination Plum Juice Concentrate 6% 4.0 4.0 4.3 4.1 Plum Juice Concentrate 8% 4.1 4.2 4.0 4.1

As described above, the single effect was examined by varying the blending ratio according to the type of additives, and based on preliminary experiments, when the appropriate concentration of plum juice was mixed at 5%, and the liquid fructose was gradually added, the overall acceptability was 7%. When glucose was added gradually, 4% was the best and there was no difference in color and aroma, but there was a change in taste preference. Also, when 8% of liquid fructose was added and then gradually added plum juice, 6% and 8% showed the same degree of synthesis and the aroma was large. In other words, the addition of liquid fructose or glucose increased the taste preference, and when plum fruit juice was added, the freshness was improved by the increase of flavor and acid taste. Therefore, from the results of sensory evaluation, the ratio of liquid fructose and glucose as sweetener was 7% and 4%, and the percentage of plum concentrate as acidic agent was in the range of 6-8%.

Experimental Example

Hereinafter, the antistress activity, antifatigue activity and the like of the yeast extract of the present invention were tested.

Experimental Example 1

Antistress activity test (1) (Kim Kyung-mi, Ph.D., Korea University)

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 ^o 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 hydrolyzed at a high temperature of 50 ° C., that is, a sample prepared in Example 1, centrifuged after hydrolysis after applying a sample 1, ultrasonic waves and vibration. 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 samples 1, 2 and 3 were lyophilized to powder and then tested.

To each male Sprague-Dawley rat, each sample (1 g / kg body weight) dissolved in distilled water was orally administered once daily 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 Table 3 below.

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 1 234.11 ± 5.31 ^b 945.18 ± 9.95 ^a 236.51 ± 29.29 7.06 ± 0.59 ^a Sample 2 243.82 ± 16.28 ^a 922.26 ± 11.93 ^a 251.43 ± 3.83 5.71 ± 0.34 Sample 3 226.43 ± 1.84 ^b 1020.18 ± 44.04 211.41 ± 10.30 5.55 ± 0.69

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

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

As a result, as shown in Table 2, when the stress (stress control) is applied, the organs that are closely related to the stress hormone production and the immune system cause significant fluctuations compared to the normal group, so that the weight of the spleen, thymus, thyroid, etc. This markedly decreased 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, although the hydrolysis of yeast by the general method and the induction of anti-stress substance secretion by ultrasonic wave and vibration were slightly different, all of them were excellent in anti-stress effect. However, the dissolution process of live yeast is thought to have the same effect as applying stress such as ultrasound or vibration.

Antistress activity test (2) (Kyung Mi Kim, Korea University)

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 Table 4 below.

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 1 182.24 ± 21.73 43.10 ± 5.09 125.0 ± 81.51 ^a 718.40 ± 59.18 ^a 15.15 ± 1.08 Sample 2 141.63 ± 3.79 ^a 31.50 ± 2.07 100.59 ± 1.56 ^b 562.40 ± 58.02 ^a 17.10 ± 0.90 ^a Sample 3 212.69 ± 10.61 55.37 ± 3.37 129.04 ± 20.87 891.07 ± 21.36 19.10 ± 1.00 ^a

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

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

As shown in Table 3 above, LDH and total cholesterol were increased by the application of stress, 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.

From the above experimental results, it can be seen that Samples 1 and 2 have excellent antistress activity, and in particular, Sample 2 has significant antistress activity.

Experimental Example 2

Anti-fatigue effect test (Kyung Mi Kim, Korea University)

In order to measure the antistress activity of the yeast extract of the present invention, anti-fatigue capacity was measured by measuring swimming endurance capacity which is closely related to the antistress factor. For the endurance of swimming, the endurance measuring device for the mouse (a water pool exercise device, Matsumoto et al., 1996) as shown in Fig. 4, which has high data reproducibility and reliability, was used. (Male ICR mice: purchased at Korea International Laboratory Animal Center) for a week, lighting time is 12 hours from 7 am to 7 pm, room temperature is 22-24 ^o C, humidity 50% (33 × 23 × 12cm, 3 heads / room) was used for this experiment. Feed was used as solid feed (purchased from CheilJedang), and water was used as tap water, and feed and water were not limited. In the control mice were orally administered a solid feed (1 g / kg), experimental mice were orally administered with the same yeast extract as prepared in Example 1, obtained by hydrolysis at a high temperature of 50 ℃ (1 g / kg) was measured three times 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 visually confirmed that the mouse is not up to the water surface until the seven seconds after falling into 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 Figure 5 below.

As shown in Figure 5, in the case of the experimental group orally administered with the yeast extract of the present invention as a sample, it can be seen that the swimming time is significantly improved compared to the control group. As a result, it was confirmed that the yeast extract of the present invention has an anti-fatigue effect and an anti-stress activity.

Experimental Example 3

Clinical Trial (Hwang Won-Joon, Neuropsychiatrist)

In order to clinically test the antistress effect on the human body of the present invention, a baseline dose is administered to a group of neuropsychiatric patients such as 1. insomnia, 2. anxiety, and 3. headache. After 3 days of brain mapping (February 6, 2001), each patient received 500 mg of the yeast extract of the present invention for 3 days a day, and then brain function test (2 2001) March 9). The results are shown in FIGS. 6A and 6B.

As shown in FIGS. 6A and 6B, the baseline test showed an asymmetric increase in almost all areas of theta. This means that mental activity is increased and mental tension is present. However, the test result after 3 days of dosing showed a symmetrical distribution in the mid-parietal region of theta. This is a state in which the state of psychological stability is achieved in a state that is seen in a relaxed state, a sleeping state or an elevation state.

The present invention reveals a novel use of yeast 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 psychotropic agent with many side effects.

In addition, the yeast peptides can be hydrolyzed by hydrolyzing food grade yeast instead of neurotrophic factor (neutrophin), which has been hardly obtained through the expression of conventional recombinant strains. It is manufactured to penetrate the cerebrovascular barrier, and as a new use of yeast extract, it has developed anti-stress effect, anti-fatigue effect, and use as a natural neurotrophic agent. Was done.

Claims (18)

Self-extinguishing the yeast at a temperature of 35 ° C. to 70 ° C .; Simultaneously or autodigesting the yeast, followed by hydrolysis of the protein with proteases; Centrifuging the hydrolyzed yeast to take only the supernatant; A yeast peptide having a molecular weight of 10,000 Da or less obtained by a process comprising the step of filtering the supernatant centrifuged using an ultrafiltration ultrafiltration membrane. The yeast peptide according to claim 1, wherein 15 to 25 mol% of glutamic acid and aspartic acid are contained. 2. The yeast peptide of claim 1, wherein the yeast peptide crosses the cerebrovascular barrier. Process for preparing yeast extract comprising the following steps: 1) selecting only the surviving strains by culturing while applying high temperature, ultrasonic wave, vibration, pH change, etc. in the range that yeast does not die; 2) culturing the selected yeast strain to the maximum growth stage; 3) self-extinguishing the cultured yeast at a temperature of 35 ℃ to 70 ℃; 4) simultaneously or after said autodigestion, further adding a proteolytic enzyme to hydrolyze the yeast protein; And, 5) centrifuging the hydrolyzed to separate only the supernatant. 5. The method of claim 4, further comprising inducing secretion of metabolites to withstand stress by applying appropriate physical or chemical stresses such as high temperature, ultrasonic waves, vibrations, pH changes, etc., to which yeast does not die before self-extinguishing the yeast. Method for producing a yeast extract, characterized in that it comprises a. A method for producing a yeast peptide comprising filtering a yeast extract obtained by the method of claim 4 or 5 using an ultrafiltration ultrafiltration membrane to take only a peptide having a molecular weight of 10,000 or less. A functional beverage comprising the step of sterilizing the yeast extract obtained by the method of claim 4 or 5 together with activated carbon in water, stirring and sterilizing the sterilized product under reduced pressure and colorlessly purifying it. Method of preparation. An antistress agent comprising the yeast peptide or the yeast extract according to any one of claims 1 to 3 or 4 or 5 as an active ingredient. 10. The formulation of claim 8, wherein the antistress agent has the activity of nerve stabilization, tension relaxation, anxiety removal or sleep assistance. An anti-fatigue formulation comprising the yeast peptide or the yeast extract according to any one of claims 1 to 3 or 4 or 5 as an active ingredient. A neuronutrient comprising the yeast peptide or the yeast extract according to any one of claims 1 to 3 or 4 or 5 as an active ingredient. An antistress, anti-fatigue and neurotrophic functional beverage comprising the yeast peptide or yeast extract according to any one of claims 1 to 3 or 4 or 5. 13. The functional beverage according to claim 12, comprising 0.1 to 10% by weight of yeast extract or yeast peptide, 10 to 25% by weight of conventional beverage additives such as sweeteners and acidulants, and the remaining weight% of water. delete delete delete delete delete