KR20160019571A - Method for producing fermented Sparassis crispa extracts with enhanced free amino acid content - Google Patents

Abstract

The present invention relates to a method for preparing a fermented Sparassis crispa extract having increased free amino acid content, compared to a Sparassis crispa extract before fermentation, wherein Aspergillus oryzae suitable for fermentation of Sparassis crispa and capable of increasing a useful ingredient of Sparassis crispa for a short time at low costs is used. According to the preparation method of the present invention, prepared is a fermented Sparassis crispa extract having an 1.1 to 1.5-fold increase in the entire free amino acid content, in particular, having 2 to 5-fold increases in cysteine and phenylalanine contents, compared to a Sparassis crispa extract before fermentation. Furthermore, the fermented Sparassis crispa extract having increased free amino acid content is easy to absorb in vivo in a human body and prevents or alleviates diseases such as bronchitis, depression, obesity, blood cholesterol, hypertension, etc. to enhance health, thereby enabling a user to be provided with a healthy food including the fermented Sparassis crispa extract.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for producing a fermented extract of Rosemary mushroom having increased free amino acid content,

More particularly, the present invention relates to a method for producing an extract of Aspergillus oryzae with an increased free amino acid content, and more particularly, to a process for producing Aspergillus oryzae by inoculating a mushroom powder of Aspergillus oryzae and fermenting the same to increase the content of cysteine and phenylalanine The present invention relates to a method for producing a fermented extract of P. falciparum.

Along with an increase in average life expectancy, food and food consumption has been moving in the direction of safety and health since 2000. As a result, organic agricultural products, non-polluting foods, functional foods, and natural foods are continuously gaining popularity. An example of such a fermented food is a fermented food prepared by adding a sugar or a microorganism. Fermented foods are fermented by decomposition of polymeric materials into low molecular weight substances to increase digestion and absorption rate, new flavor is generated by the creation of palatability, as well as increased physiologically active ingredients, in addition to drugs for the maintenance and treatment of health in general, Research on the effect of physiological activity on ingestible substances has been extensively conducted, and functional foods and ancillary medicines have been actively developed (Heo SI et al. , 2010).

Sparassis crispa is an edible mushroom belonging to the genus Rosaceae. It is rich in dietary fiber, especially beta-1, 3-linked beta-1, 6-linked beta-1, It is known that the content of glucan (β-glucan) is more than 10 times as much as that of the edible mushroom or mushroom. The above-mentioned beta-glucan promotes the production of interleukin and interferon which inhibits proliferation and recurrence of cancer cells and activates immune cell function, and thus provides antitumor, antitubercity, antidiabetic, antioxidant, blood cholesterol lowering, It is known to have various physiological activities such as anti-inflammation.

These mushrooms can be consumed directly as food, but they are limited to ingesting unprocessed foodstuffs in order to take a pharmacological effect. In addition, when raw juice is directly consumed, the amount of water absorbed is small and impurities are large. Thus, methods for extracting useful components and taking them for a long time have been studied. However, it is necessary to develop new type of product that can show the functional effect on ingestion because it is difficult to extract the useful ingredient of the mushroom and it is difficult to absorb in the body only by hot water extraction.

Korean Patent Laid-Open Publication No. 2011-0082758 discloses a health supplement product using an extract of Zygomy mushroom which is obtained by fermenting Zygomaster mushroom with a microbial solid fermentation, Discloses a method for producing fermented food of lactic acid bacteria mixed with mushroom and brown rice, and Korean Patent Publication No. 2010-0112054 discloses a method for producing a fermented product containing vegetable extract and dried mushroom as main components. However, the microorganisms used for fermentation in the above-mentioned cited documents are expensive because they use mixed microorganisms, and the lactic acid bacteria have a disadvantage that they can not form spores, so that stability against heat, acid, oxygen and the like is lowered. The fermented mushroom extract has a problem in that the content of useful components is insufficient.

Accordingly, the present inventors have found that a fermented microorganism capable of enhancing the content of useful components of Zanthoxylum mushroom in a short period of time can be selected and inoculated into Zanthoxylum mushroom to obtain a fermented extract of Zanthoxylum mushroom having an increased free amino acid content such as cysteine and phenylalanine As a result of efforts to develop, the present invention has been completed.

It is an object of the present invention to provide a method for producing a fermented extract of P. falciparum with an increased content of free amino acid using Aspergillus oryzae .

Another object of the present invention is to provide a fermented mushroom fermented extract prepared by the above production method, wherein the content of free amino acid is increased to facilitate absorption in the human body, improve health by eliminating obesity, lowering cholesterol in blood and preventing hypertension And a health food containing the fermented extract of Mushroom mushroom.

The present invention relates to a method for producing a mushroom fermented extract having an increased free amino acid content by inoculating mushroom powder into a mushroom powder and then fermenting the mushroom powder.

Fermentation is accomplished by decomposing organic matter using enzymes possessed by microorganisms to produce useful substances or converting them into components that are easily absorbed into human cells. The fermenting microorganism has a property of fermenting food and can be safely used. Examples thereof include lactic acid bacteria, acetic acid bacteria, Bacillus subtilis , butyric acid bacteria, hiochi bacteria, fungus and yeast.

However, since these fermenting microorganisms produce different substances depending on the source of fermentation and, in some cases, produce harmful substances to human body, it is important to select useful fermenting microorganisms according to fermentation source and purpose of fermentation.

Accordingly, the present inventors have found that it is possible to produce a useful substance in a short period of time while being suitable for fermentation of Zygomycetes mushroom among various fermenting microorganisms described above, and to increase an anti-obesity effect, an increase in blood cholesterol lowering effect, And fermentation microorganisms which may result in the increase of microorganisms.

As a result, in the present invention, Aspergillus oryzae was found as a fermenting microorganism.

In one specific embodiment, Aspergillus duck claim (Aspergillus oryzae) in the case of vaccination to the fermentation was prepared blossoms mushroom fermentation extract, but the amount of the total free amino acids as compared to non-fermented blossoms mushroom extract increased, Pseudomonas coarse louver (Monascus The content of total flavonoids and total free amino acids was decreased in the fermented extract of P. japonicus prepared by fermentation with inoculated ruber .

In the present invention, the free amino acid may be at least one selected from the group consisting of cysteine, serine, glycine, alanine, tyrosine, and phenylalanine, Is cysteine or phenylalanine. In particular, the phenylalanine is an essential amino acid and is not synthesized in the human body and is an amino acid that should be ingested as a food. Therefore, the extract of Zygomy mushroom fermented with increased phenylalanine content can be more usefully used.

The cysteine is one of the alpha amino acids and is an essential amino acid, but it is an essential amino acid for infants, the elderly, people with characteristic metabolic diseases, or persons suffering from malabsorption syndrome. These cysteines, along with aspartic acid and citrulline, are necessary to detoxify toxins produced by alcohol ingestion and smoking, to maintain collagen production and skin elasticity, and to prevent damage from radiation. In addition, it is known that it has a therapeutic effect on bronchitis, emphysema, and tuberculosis because it has the ability to prevent the generation of coloring matter such as black pigment of the skin, burns body fat, and removes mucus from the bronchial tube.

The serine is an amino acid biosynthesized from glycine. It helps the production of immunoglobulins and antibodies, maintains a healthy immune system, and helps muscle growth and fat metabolism. It is also known to be used as a raw material for a natural wetting agent, which is a skin protecting agent in clinical use, and is used for treatment necessary for improving immune function.

The glycine is an amino acid that assists in the synthesis of other aminic acid. It is essential for the biosynthesis of collagen. It acts as an antacid to help treat gastric hyperplasia and removes free radicals from the liver. It is also known to be used for the treatment of progressive muscular stiffness, epileptic seizures, depression, hypoglycemia, prostate disorders, gastric hyperplasia, hepatic diseases, muscle and cartilage degeneration in the clinic.

Alanine promotes hepatic metabolism, detoxifies harmful substances, helps alcoholic enzymes act actively, and plays a role in quickly eliminating alcohol by diuretic action. In addition, it is known that it not only regulates the energy of glucose, but also regulates blood sugar and also acts on the enlarged prostate.

Tyrosine is one of the alpha amino acids present in the thyroid gland. Tyrosine can help the brain function to relieve depression, increase the amount of neurohormones, and treat Parkinson's disease. Tyrosine is also known to be effective in the treatment of hypotension and hypothermia as well as helping obesity by reducing appetite and reducing body fat.

The phenylalanine, one of the essential amino acids, passes through the blood brain barrier and acts directly on the chemicals in the brain. It also has excellent antidepressant action, delaying the breakdown of physiologically active analgesic substances, prolong analgesic efficacy, and control inflammation. In addition, it is known that it enhances memory and learning ability, inhibits appetite, and is also effective in the treatment of diseases such as arthritis, migraine and Parkinson's disease.

In the present invention, the increase in the content of free amino acids means that the total amino acid content is increased by 1.1 to 1.5 times as compared with the extract before fermentation. In particular, the content of cysteine and phenylalanine is 2 to 5 times, 4-fold and phenylalanine increased 3 to 5 times.

In one specific embodiment, the fermented extract of Aspergillus oryzae ( Aspergillus oryzae ) was inoculated with 2% of the extract of Aspergillus oryzae and fermented by extracting and concentrating 2% of the extract of Aspergillus oryzae and the free amino acid Was measured. As a result, the total free amino acid content of the fermented extract of P. japonica increased about 1.2 times as compared with that of the unfermented P. japonicus extract. Specifically, the contents of cysteine, serine, glycine, threonine, alanine, tyrosine and phenylalanine in the free amino acids were increased , Especially cysteine and phenylalanine, were increased about 3.7 times and 4.8 times, respectively, as compared to the non - fermented.

In the present invention, the mushroom powder is a powdery material obtained by washing, drying and crushing the mushroom. The drying is not limited as long as it is a method of removing water from the mushroom. For example, natural drying, hot air drying, vacuum drying and the like can be used. There is a problem that if the moisture on the surface of the mushroom is not removed, the mushroom of the mushroom does not recede or break up when it is cut. The pulverization is preferably performed at 50 to 150 mesh, preferably 70 to 120 mesh, and the number of pulverization is preferably 1 to 3 cycles.

In the present invention, it is preferable that the Aspergillus oryzae is inoculated in an amount of 0.5 to 5% by weight, preferably 1 to 3% by weight based on the total weight of the mushroom powder. If the content of Aspergillus oryzae is less than 1% by weight, the fermentation may not occur uniformly and the quality of the mushroom may be deteriorated. If the content of Aspergillus oryzae is less than 5% The fermentation efficiency is not increased as compared with the case where the amount is less than the above amount, which is uneconomical.

In the present invention, Aspergillus oryzae can be added to the strain itself or a culture thereof. As a specific example, the culture medium of Aspergillus oryzae refers to a culture solution in which the number of bacteria is increased by inoculating each strain into a subculture medium. As an example, 10 ml of high pressure sterilized PDB medium (200 g / L of potato starch, 20 g / L of dextrose) was inoculated with an Aspergillus oryzae strain and incubated at 25-40 ° C for 30-48 hours at 200 rpm And may be obtained by stirring.

In the present invention, the fermentation is carried out at a temperature of 25 to 40 占 폚, preferably 28 to 35 占 폚, for 5 to 15 days, preferably 7 to 12 占 폚, at a temperature of 25 to 40 占 폚 for Aspergillus oryzae- It can be done for days. If the fermentation temperature is less than 25 ° C or the fermentation time is less than 5 days, the useful ingredient may remain in the mushroom. If the fermentation temperature exceeds 40 ° C or the fermentation time exceeds 15 days, to be.

In the present invention, the fermentation extract of P. japonicus can be obtained by using an extraction method and an extraction solvent known in the art, preferably a hydrothermal, an anhydrous or a functional lower alcohol having 1 to 4 carbon atoms , Ethanol, propanol, butanol, etc.), acetone, ethyl acetate, chloroform, and 1,3-butylene glycol may be used singly or in combination of two or more. More preferred are anhydrides or lower alcohols having 1 to 4 carbon atoms And still more preferably methanol or ethanol. The extraction method is not particularly limited, and examples thereof include cold extraction, ultrasonic extraction, and reflux cooling extraction.

In addition, the above-mentioned fermentation extract of P. japonicus can also be obtained by a conventional purification process in addition to the above extraction method. For example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatographies (made for separation by size, charge, hydrophobicity or affinity) An extract of P. falciparum fermented by the obtained fraction can also be obtained.

In the present invention, the Fusarium oxysporum fermentation extract may be any extract, fraction and purified product obtained in each step of filtration, extraction, fractionation or purification, a diluted solution thereof, a concentrate or a dried product thereof.

On the other hand, the health food containing the fermented extract of Zygomy mushroom produced according to the present invention can improve health by improving obesity, lowering of cholesterol in blood, prevention of hypertension and the like.

As a specific example, the present invention can be used as an additive to a food product. According to the results of the present inventors' study, the total content of free amino acids in the fermented extract of P. japonicum according to the present invention is 1.1 to 1.5 times The content of cysteine, serine, glycine, threonine, alanine, tyrosine and phenylalanine in the free amino acids was increased, Especially, cysteine was about 3.7 times higher than that of the non-fermented mushroom extract and phenylalanine was about 4.8 times higher than that of the non-fermented mushroom extract. Therefore, the amount of free amino acid increased in the food industry Mushrooms are easy to absorb in the body, and prevent bronchial disease, anti-depression, obesity improvement, And lowering of blood cholesterol, prevention of hypertension, and the like.

In the present specification, the term "health promotion" means prevention or amelioration of diseases such as bronchitis, depression, obesity, blood cholesterol, and hypertension, thereby improving nutrition and fitness.

In the present invention, when the fermented extract of Zanthoxylum mushroom is used for food, the fermented extract of the present invention may contain components that are usually added during the manufacture of food. For example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. The above-mentioned carbohydrates are polysaccharides such as disaccharides such as monosaccharides of glucose and fructose sugars, maltose, sucrose and oligosaccharides, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. The flavoring agent may be a natural flavoring agent (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavorings (saccharin, aspartame, etc.).

There is no particular limitation on the kind of the food. Examples of the food to which the above-described mushroom fermentation extract can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, , Drinks, alcoholic beverages, and vitamin complexes, all of which include health supplements in the usual sense.

When the food of the present invention is a beverage, it may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame, and the like .

The food may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These components may be used independently or in combination.

The present invention relates to a method for fermenting a mushroom of Aspergillus oryzae which is suitable for fermentation of a mushroom and can increase a useful ingredient of the mushroom by a small amount of time in a short time, Wherein the content of total free amino acids is increased by 1.1 to 1.5 times, and especially the content of cysteine and phenylalanine is increased by 2 to 5 times. In addition, the fermented mushroom fermented extract prepared by the above-mentioned method has an increased free amino acid content and is easily absorbed into the human body and can prevent or improve diseases such as bronchitis, depression, obesity, blood cholesterol and hypertension, Therefore, it is possible to provide a health food containing the fermented extract of Mushroom mushroom.

Fig. 1 shows the results of comparative measurement of the total polyphenol contents of the mushroom fermented mushroom extract prepared by inoculation with Monascus ruber or Aspergillus oryzae and the unfermented mushroom extract.
FIG. 2 shows the results of comparative measurement of the total flavonoid content of the fermented mushroom fermented extract prepared by inoculation with Monascus ruber or Aspergillus oryzae and the unfermented mushroom extract.
FIG. 3 shows the results of comparison of the free amino acid contents of the mushroom fermented mushroom extract prepared by inoculation with Monascus ruber or Aspergillus oryzae and the unfermented Mushroom mushroom extract.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, and it is to be understood by those skilled in the art that the present invention is not limited thereto It will be obvious.

Preparation Example 1: Culture of strain and preparation of strain

1-1. Production of Monascus ruber culture

Monascus ruber purchased from Korean Microorganism Conservation Center was inoculated into a PDA (potato dextrose agar) medium and streak cultured at 30 ° C for 7 days to produce spores. Next, 1 ml of Tween 20 was dropped on the medium to take spores. Then, 100 ml of PDB (potato dextrose broth) was inoculated and cultured at 30 ° C for 7 days with shaking to form mycelia. The mycelium was then aseptically ground and suspended in saline.

800 mg of brown rice was added to 100 ml of distilled water, sterilized at 121 ° C for 15 minutes, cooled, 10 ml of the monascus luer mycelium was inoculated and cultured at 30 ° C for 10 days to prepare a Monascus lucer culture.

1-2. Preparation of Aspergillus oryzae

Aspergillus oryzae purchased from the Korean Microorganism Conservation Center was inoculated into a PDA (potato dextrose agar) medium and streak cultured at 30 ° C for 7 days to prepare spores.

Example 1: Preparation of Fermented Extract of Mushroom

The mushroom was washed three times with running water, and then dried in a 1m × 1m dried wipe to remove water. And then cut to a thickness of 5 mm or less using three seasons. Then, the above-mentioned three fine-grained mushrooms were dried for 24 hours and then subjected to primary pulverization (control), secondary pulverization (turmeric) and third pulverization (seeding), and then passed through a 100-mesh sieve to prepare powdery mushroom powder . Then, 100 g of the above mushroom powder was prepared in Preparation Example 1, and 2 ml of a Monascus ruber culture solution or 2 ml of Aspergillus oryzae strain was inoculated, followed by fermentation at 30 ° C for 10 days. Then, 80% methanol was added to the fermented Rosemary mushroom, extracted with an ASE accelerator, and then concentrated using a rotary vacuum concentrator to prepare a fermented extract of P. falciparum.

Example 2: Determination of the content of useful components of the fermented extract of Zanthoxylum mushroom

2-1. Total Polyphenol Content of Fermented Extracts from Mushroom Mushroom

2 ml of sodium carbonate was added to 1 g of the mushroom fermented extract fermented with Monascus ruber or Aspergillus oryzae in Example 1, and then 50% of Folin-Ciocalteu reagent reagent) was added. The absorbance was then measured at 720 nm using a spectrophotometer and the total polyphenol content of the fermented extract of P. falciparum was determined based on the calibration curve of gallic acid (Sigma Co., USA). As a control group, the non - fermented Zygomycota mushroom extract was used. The results are shown in Fig.

As a result, the total polyphenol content of the fermented extract of Aspergillus oryzae fermented with Aspergillus oryzae was 8.04 ± 0.87, and the total polyphenol content of the Fusarium mushroom extract fermented with Monascus ruber was 6.74 ± 0.85 and the fermented extract of Aspergillus oryzae showed a slightly higher total polyphenol content than that of Monascus ruber but not significant difference. The total polyphenol content of the non - fermented Zygomyksis mushroom extract was 7.94 ± 1.70, which was higher than that of Monascus ruber .

2-2. Determination of total flavonoid content of fermented Extract of Mushroom Mushroom

10 ml of diethylene glycol was added to 1 g of the mushroom fermented extract fermented with Monascus ruber or Aspergillus oryzae in Example 1 and mixed with 1 ml of 1N NaOH The mixture was incubated at 37 ° C for 1 hour, absorbance was measured at 420 nm, and the total flavonoid content of the mushroom fermented extract was measured based on the calibration curve of rutin (Sigma Co., USA). As a control group, the non - fermented Zygomycota mushroom extract was used. The results are shown in Fig.

As a result, the total flavonoid content in the fermented extract of Aspergillus oryzae fermented with Aspergillus oryzae was 16.13 ± 4.12, and the total polyphenol content of the extract of Fusarium oxysporum fungus fermented with Monascus ruber was 10.63 ± 0.52. The total fermented extract of Aspergillus oryzae showed slightly higher total flavonoid content than that of Monascus ruber fermented by Monascus ruber , but showed no significant difference. The total polyphenol content of the non - fermented Zygomyksis mushroom extract was 15.46 ± 2.92 and the total flavonoid content was higher than that of the Zygomy mushroom extract fermented with Monascus ruber .

2-3. Determination of Free Amino Acid Content of Fermented Extracts from Mushroom Mushroom

In Example 1, 0.2 g of fermented Mushroom fermented by fermentation with Monascus ruber or Aspergillus oryzae was added to 60 ml of ethanol to extract amino acids, and then other ingredients such as fat were removed by ether And concentrated under reduced pressure in a constant temperature water bath at 55 ° C. The concentration of the concentrated fermentation extract was adjusted to 25 ml using a pH 2.2 diluted solution, and the free amino acid content and content thereof were analyzed using an amino acid analyzer (Biochrom, UK). As a control group, the non - fermented Zygomycota mushroom extract was used. The results are shown in Table 1 and FIG.

Free amino acid component
(ng / mg ext.) Control group Aspergillus orije
( Aspergillus oryzae ) Monas Kerr Louver
( Monascus ruber ) Cysteine 1.014.04 3,776.29 1,582.80 Asp ¹⁾ 5,158.03 6,618.75 9,206.22 Glu ²⁾ 9,525.86 7,772.63 224.60 Serine 1,669.66 1,828.76 7.69 Glycine 2,052.18 2,531.30 34.15 Histidine 750.00 660.17 20.35 Arginine 1,287.71 1,104.56 37.20 Threonine 1,588.90 1,653.62 37.70 Alanine 2,694.21 3,008.68 10.66 Proline 1,924.73 1,976.46 21.30 Tyrosine 476.80 517.55 53.95 Valine 1,909.24 1,546.82 74.47 Methionine 212.96 186.90 27.43 Isoleucine 1,239.04 1,082.24 4.65 Leucine 3,204.38 2,265.21 - Phenylalanine 1,034.28 4,993.10 - Tryptophan 641.30 666.58 459.73 Lysine 1,117.38 1,367.05 31.10 Sum 37,500.70 43,556.67 11,834.03 ¹⁾ a component in which aspartate and asparagine contents are added, ²⁾ a component in which glutamate and glutamine contents are added

As a result, the free amino acid content of the fermented extract of Aspergillus oryzae fermented by Aspergillus oryzae was the highest at 43,556.67 ng / mg, which was higher than that of the non - fermented Zygomyx mushroom extract. Specifically, the contents of cysteine, serine, glycine, threonine, alanine, tyrosine and phenylalanine in the free amino acids were increased, especially cysteine ) Was about 3.7 times higher than that of the unfermented mushroom extract and phenylalanine was about 4.8 times higher than that of the unfermented mushroom extract.

However, the free amino acid content of the mushroom extract fermented with Monascus ruber was 11,834.03 ng / mg, which was lower than that of the mushroom extract which was not fermented.

Claims (4)

A method for producing a fermented extract of P. falciparum mushroom having an increased free amino acid content by inoculating fungus with a fungus powder and then fermenting at a temperature of 25 to 40 ° C for 5 to 15 days to extract and concentrate the fungus, Wherein the fermentation is carried out by inoculating Aspergillus oryaze in an amount of 0.5 to 5% by weight based on the total weight of the mushroom powder, and the fermented mushroom fermentation extract of Aspergillus oryzae Wherein the total free amino acid content is increased by 1.1 to 1.5 times as compared to the non-fermented Zanthoxylum mushroom extract.
The method according to claim 1, wherein the free amino acid is at least one selected from the group consisting of cysteine, serine, glycine, alanine, tyrosine, and phenylalanine A method for producing a fermented mushroom extract.
The method according to claim 1, wherein the free amino acid is cysteine or phenylalanine.
A health promoting food for preventing or improving bronchitis, depression, obesity, blood cholesterol, or hypertension, including the fermented extract of Mushroom mushroom produced by the production method of any one of claims 1 to 3.

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