KR20190084592A - Manufacturing method of natural fermented-composition having fixed nitric oxide and natural fermented-composition thereof - Google Patents

Abstract

The present invention relates to a method of producing a naturally fermented composition for fixing nitrogen oxide in a naturally fermented substance, and to a naturally fermented composition produced thereby. More specifically, the present invention provides a natural product-based NO producing preparation which stably contains a large quantity of nitrogen oxide products to promote the maintenance of a normal NO balance in a human body, is convenient to ingest, and does not cause adverse side effects. To this end, according to one embodiment of the present invention, the method mixes a nitrogen-containing organic substance existing in the nature, water, and a fermentation bacteria strain and ferments the mixture at room temperature for 5-30 days, thereby causing sugars and proteins of the organic substance to be disintegrated and converted; propagates the fermentation bacteria strain and increase a distribution ratio thereof, thereby maximizing ammonia formation; supplies additional carbon sources; converts ammonia nitrogen in filtrate to nitrites; and fixes residual organic matters by causing the same to be disintegrated and converted to stable R-NO and R-S-NO compounds.

Description

Technical Field [0001] The present invention relates to a method for preparing a natural fermentation composition for immobilizing nitrogen oxide in a natural fermentation material and a natural fermentation composition prepared by the method.

The present invention relates to a method for producing a natural fermentation composition for immobilizing nitrogen oxide in a natural fermentation material and a natural fermentation composition prepared by the method. More particularly, the present invention relates to a method for producing a natural fermentation composition, To a natural fermentation composition containing a large amount of fixed nitrogen oxide.

Nitric oxide (NO) is a very short gas molecule with a half-life of several seconds. It plays a vital role in maintaining homeostasis of the body by participating in vasodilatation, neurotransmitter and immune function.

When the normal balance of nitrogen oxides breaks down, it exerts a great influence on various pathological diseases such as cardiovascular diseases such as atherosclerosis, respiratory diseases, platelet dysfunction, immune dysfunction, Alzheimer's disease and sexual dysfunction.

Nitrate (NO ₃ ^- ) and nitrite (NO ₂ ^- ) can be converted to nitric oxide by reduction reaction with nitric oxide. Nitrate is reduced to nitrites by bacteria in the mouth. Nitrite is reduced by acid to form nitrogen oxide.

Table 1 summarizes the functions and possible clinical applications of representative nitric oxide in human body.

Nitric oxide
function Explanation Clinical application Cardiovascular regulation Nitric oxide is a powerful blood product in the body that improves blood flow and regulates blood pressure. Hypertension, pulmonary hypertension, sexual dysfunction, diabetes, arteriosclerosis Inflammation and immune regulation Nitric oxide is secreted from activated immune cells, killing pathogens such as bacteria and viruses, preventing leukocyte adhesion and suppressing inflammatory responses. Antibacterial, wound healing Neurotransmitter It acts as a neurotransmitter between neurons and is easy to diffuse into the environment due to fat-solubility and free from intracellular inflow. Alzheimer's Anticoagulation reaction Platelet aggregation inhibition. Prevents restenosis and inhibits thrombus formation stent coating, various graft coating

Most of the above diseases are caused by lack of nitrogen oxides, and there are a lot of studies to treat diseases by restoring normal nitric oxide concentration.

For this purpose, there have been studies to increase NO production by activating nitric oxide synthase (NOS) or by administering a substance (eg, L-arginine) that is involved in NO production.

Nitric oxide synthase produces nitrogen oxides by oxidizing five electrons of guanidino nitrogen of L-arginine. However, the results of clinical trials have failed. The reason for this is that most people with low levels of nitric oxide levels are suffering from endothelial dysfunction and lacking the ability to convert NO. It is also because of the stiffness of the blood vessels. Particularly, at the age of 40 years or older, the activity of nitric oxide synthase is lowered, and the portion contributing to nitric oxide production of L-arginine is further reduced. Thus, nitric oxide supplements based on L-arginine have definite limitations on their function.

On the other hand, there is a way to heal illness by directly supplying deficient NO from the outside. That is, NO itself is used as a therapeutic drug. In fact, NO gas is a lung disease drug such as pulmonary hypertension is directly inhaled gas treatment is being used. However, NO is gas, strong reactivity, and most of all has a very short half-life (2 to 4 seconds), which limits its clinical application.

Various NO donors have been developed that can store and release NO. Representative NO donors include isosorbide mononitrate and nitroglycerine, which are used in angina pectoris, heart failure, and the like. However, these donors have side effects such as hypertension, headache, and tolerance, making them difficult to use for other diseases.

Recent nitrosoglutathione donors include diazenium diolate (NONOate) and RSNO, an endogenous NO donor such as S-nitrosoglutathione. These donors are widely used because they can store and release NO more stably than the previous donors.

However, these NO donors also have various release mechanisms, and they are particularly sensitive to environmental changes such as various enzymes and pH in the human body, so that the control of NO release is very difficult, and there is a concern about side effects because there is no specificity for disease sites. Therefore, studies have been actively conducted recently to make available a safe and sufficient amount of NO in the body more efficiently.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a naturally- NO production formulations.

According to an aspect of the present invention, there is provided a method of preparing a natural fermented composition for immobilizing nitric oxide in a natural fermenting material, the method comprising: mixing a nitrogen-containing organic substance present in a natural environment, , Forest products, aquatic products, minerals, water, and fermentation strains, and then fermented at room temperature for 5 to 30 days to decompose and convert sugar and proteins of the organic material. Ammonia; Converting the carbon source and the oxygen supply to the fermentation product to convert the ammonia nitrogen of the filtrate into nitrite nitrogen and decomposing the remaining organic substances of the nitrite nitrogen and converting and fixing the R-NO and RS-NO to the nitrite nitrogen .

Here, the amount of water is 1 to 20 times (weight ratio) of the nitrogen-containing organic material and the amount of the fermenting bacteria is preferably 0.01 to 1 part by weight with respect to 100 parts by weight of the mixture of the nitrogen-containing organic material and water.

In addition, the fermenting bacteria may be Bacillus subtilis subsp. Subtilis (microorganism deposit number KACC 91554P).

In addition, the above manufacturing method further includes the steps of converting nitrite nitrogen into R-NO and RS-NO and immobilizing the nitrite nitrogen for 24 to 36 hours in an anoxic state to stabilize the immobilized nitric oxide and precipitating the remaining suspended substances can do.

In addition, the above manufacturing method may further include an ultrafiltration step of removing nitrite nitrogen from the fermentation product after the conversion and immobilization of R-NO and RS-NO, followed by a sterilization or sterilization step .

In order to achieve the object of the present invention, the natural fermentation composition according to another aspect of the present invention is characterized in that it is prepared by the above-mentioned production method.

The natural product-based oxidative gas producing agent according to the present invention decomposes the remaining organic matter of nitrite nitrogen (NO ₂ ^- ), which is a precursor of nitric oxide, into a form of R-NO and RS-NO, In order to maintain the effect.

In addition, the natural product-based oxidative gas producing agent according to the present invention has an advantage in that the quality is not deteriorated even when stored for a long period of time because it is stably maintained in the contained fermentation material.

In addition, the natural product-based oxidative gas producing agent according to the present invention is a fermentation product based on a natural substance, which is filled with the amount of nitrogen oxides which are insufficient in the human body regardless of the function of the endothelial cells, and has no side effects and no resistance.

In addition, the natural product-based oxidative gas producing agent according to the present invention can be developed into a variety of materials such as liquid, powder, and capsule, and has an advantage that it is easy to ingest, is quick to absorb, and has an immediate effect.

1 is a view showing a process for producing a natural fermentation composition according to the present invention.
2 (a) and 2 (b) are tables summarizing the results of measurement of the nitrate (NO ₃ ^- ) and nitrite (NO ₂ ^- ) contents of the natural fermentation composition according to the present invention every five months.
FIG. 3 is a comparison chart showing the ability of the natural fermentation composition according to the present invention to produce nitric oxide from a commercially available nitric oxide supplement.
FIG. 4 is a graph showing the amount of nitrogen oxide released over time in the fermented garlic composition according to the present invention and commercially available Neo40 (manufactured by Neogenesis Lab). FIG. 4 (a) is a graph showing the amount of nitrogen oxide released over time ) Is a graph showing cumulative total amount of nitrogen oxides emitted over time.
5 is a nitrogen cycle diagram.

Hereinafter, a method for immobilizing a nitric oxide-based nitrification according to the present invention will be described in detail with reference to FIG.

(1) Step 1

The nitrifying nitrification fermentation method supplies oxygen and organic degrading microorganism strains to decompose the organic substances of the fermentation substance.

Nitrogen-containing organic substances (agricultural products, livestock products, forest products, aquatic products, etc.) present in the natural environment are fermented after fermentation of the fermentation strains to decompose and convert organic substances and proteins in agricultural, fish, ) Ratio, thereby increasing the distribution ratio of the bacteria. That is, inducing the proliferation of the fermenting bacteria with a lowered C / N ratio.

The fermenting bacteria are not limited as long as they are fermented with nitrogen-containing substances (agricultural products, livestock products, forest products, aquatic products, etc.) and are involved in nitrification. For example, a Bacillus strain can be used. In the present invention, Bacillus subtilis subsp. Subtilis (microorganism deposit number KACC91554P) was used.

Concentrated freshwater fish, water, and fermentation strains are added to the reaction tank and run for 5 to 30 days at room temperature. The operation time can be flexibly operated according to the concentration change of the nitrogen-containing organic substance. The amount of water is 1 to 20 times (by weight) concentrated aqueous fishery product to be administered, and the amount of the fermenting microorganism is preferably 0.01 to 1 part by weight based on 100 parts by weight of the mixture of concentrated fishery product and water.

The organic matter of agriculture, fish, shrimp, fish and marine products is divided into the degradable substrate and the substrate which is slowly biodegraded.

If the temperature increased during fermentation to 20 ~ 25 ℃, set the amount of ammonia nitrogen to required oxygen consumption as the design value, 500mg / L ~ 5000mg / L depending on the substance and continue fermentation. At this time, the fermented product can be fermented as it is or after solid-liquid separation. Oxygen supplied in this process is mostly used for ammonia oxidation and has a great influence on nitrite conversion.

(2) Step 2

(NH ₄ ⁺ -N) is converted into nitrite nitrogen (NO ₂ ^- ), which is a precursor of nitric oxide, by decomposing the remaining organic substances of the nitrite nitrogen, -NO < / RTI >

This is the step for ammonification and nitrification of fermentation broths, shrimps, forests, aquatic products, and nitric oxide immobilization by fermentation strains. Fermentation is completed when the nitrite nitrogen (NO ₂ ^- ) in the fermentation product is converted and fixed to R-NO and RS-NO.

At this time, the converted nitric oxide precursors are immobilized and converted into nitric oxide in gastric juice condition or vascular endothelial cell and vitamin mixing condition (or hypoxic condition) for drinking.

(3) Step 3

Nitrite nitrogen is converted and immobilized in R-NO and R-S-NO, and then left in anoxic condition for 24 to 36 hours to stabilize the immobilized nitric oxide and precipitate the remaining suspended substances. The fermented product after this step is subjected to centrifugation and filtration. A sterilization or sterilization process may be added for long-term distribution. In the case of ultrafiltration, microorganisms are filtered, so that the process of sterilization or sterilization may be omitted.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. It will be apparent to those skilled in the art, however, that the following examples are illustrative of the present invention only and do not limit the scope of the present invention. That is, a simple modification or change of the present invention can be easily performed by those skilled in the art, and all such modifications and alterations can be considered to be included in the scope of the present invention.

Example 1 Preparation of Garlic Fermentation Composition

2 g of Bacillus subtilis subsp. Subtilis (microorganism deposit number KACC91554P) was added to 10 kg of water at 25 ° C and 1 kg of raw garlic ground, and aerobic fermentation was performed at room temperature. When the temperature was elevated during the fermentation period, it was confirmed that the temperature was lowered to 20 ~ 25 ° C after 20 days, and then the fermentation product was subjected to solid-liquid separation to obtain a filtrate. The filtrate was aerobically fermented at 25 DEG C for 5 days. To the filtrate, 50 g of pulverized crust was added as a carbon source necessary for removing ammonia, and after the third fermentation at 25 ° C for 12 days, the fermentation broth was filtered through a commonly used ultrafiltration membrane to complete the fermented garlic solution of the present invention.

Example 2 Preparation of Yeast Fermentation Composition

The other procedures are the same except that raw livestock is used instead of garlic as a main ingredient in comparison with Example 1.

Example 3: Preparation of a macromolecule fermentation composition

The other procedures are the same, except that the raw material was used instead of garlic as the main material as compared with Example 1. [

Example 4 Production of Dried Fermented Fermentation Composition

The other procedures are the same, except that the main raw material was used instead of garlic as the main raw material.

≪ Example 5 > Preparation of fermented composition of Rhizosphere

The other procedures are the same except that the raw fish was used instead of garlic as a main ingredient in comparison with Example 1.

≪ Example 6 > Production of Aronia fermentation composition

The other procedures are the same except that, as compared with Example 1, the raw material was used as the raw material instead of garlic.

Example 7 Preparation of Onion Fermented Composition

Other procedures are the same except that raw onion was used instead of garlic as a main ingredient as compared to Example 1. [

≪ Example 8 > Preparation of Fermentation Fermentation Composition

The other procedures are the same, except that the fresh juice was used instead of garlic as the main ingredient as compared to Example 1. [

≪ Example 9 > Preparation of fermented fermented composition

The other procedures are the same except that raw fermentation was used instead of garlic as the main ingredient as compared to Example 1.

Example 10: Preparation of a fermented composition of nominal rhizome (compound herb medicine)

The other procedures are the same except that nominal rhizomes were used instead of garlic as the main ingredient as compared to Example 1.

Example 11 Preparation of Garlic Lettuce Fermentation Composition

2 g of Bacillus subtilis subsp. Subtilis (microorganism deposit number KACC91554P) was added to 10 kg of water at 25 캜, 1 kg of raw garlic grounds and 2 kg of lettuce, and aerobic fermentation was carried out at room temperature for 25 days. To the fermentation broth was added 50 g of ground crust as a carbon source necessary for ammonia removal, followed by further fermentation at room temperature for 12 days. The fermentation broth was allowed to stand in anoxic condition for 30 hours, centrifuged to separate solids, the filtrate was filtered through an ultrafiltration membrane, and the filtrate was concentrated to a volume of 5% to complete the present invention.

Example 12: Preparation of a fermented composition of garlic green bean sprout

Other procedures are the same except that 1 kg of raw garlic ground powder and 1.5 kg of green bean sprouts are used as main ingredients as compared with Example 11.

Example 13: Preparation of fermentation composition of garlic Saururus chinensis

The other procedures are the same except that 1 kg of raw garlic ground powder and 0.5 kg of dried green tea were used as main ingredients as compared with Example 11.

Example 14 Preparation of Fermented Garlic Cabbage Composition

The other procedures are the same except that 1 kg of raw garlic ground powder and 2 kg of cabbage were used as main ingredients as compared with Example 11.

Example 15: Preparation of fermented composition of bean sprouts in garlic

Other procedures were the same except that 1 kg of raw garlic ground powder and 1.5 kg of ground bean sprouts were used as the main material as compared with Example 11.

<Example 16> Preparation of fermented composition of garlic king mushroom

The other procedures are the same except that 1 kg of raw garlic ground powder and 0.3 kg of Daejung mushroom were used as main ingredients as compared with Example 11.

Example 17 Preparation of Fermentation Composition of Garlic, Bean Sprouts, Green Tea, Mulberry Leaf, and Fig Leaves

As compared with Example 11, the other procedures are the same except that 1 kg of raw garlic ground powder, 0.2 kg of soybean sprouts, 0.2 kg of green tea, 0.2 kg of mulberry leaves and 0.2 kg of fig leaves are used as main ingredients.

<Experimental Example 1>

The nitrate (NO ₃ ^- ) and nitrite (NO ₂ ^- ) contents of the natural fermentation compositions prepared in Examples 1 to 17 were measured. We also examined changes in these components over a period of 5 months. The results are shown in Fig.

From the measurements in FIG. 2, it can be seen that the content of nitrate (NO ₃ ^- ) and nitrite (NO ₂ ^- ) contained in each fermentation composition is maintained very steadily.

<Experimental Example 2>

A single intake of the garlic fermented composition prepared in Example 1 and a commercially available nitric oxide supplement was added to a solution (75 mM NaCl, 13 mM KCl, 0.01 M KCl, pH 2) The amounts were compared and compared.

FIG. 3 is a comparison chart showing the nitric oxide production ability of garlic fermented composition according to the present invention and commercially available Neo40 (manufactured by Neogenesis Lab), Proargi9 + (manufactured by Synergy worldwide) and Niteworks (manufactured by Herbalife).

In Figure 3, &quot; measurement &quot; represents the maximum cumulative measurement that is emitted.

FIG. 4 is a graph showing the amount of nitrogen oxide released over time in the fermented garlic composition according to the present invention and commercially available Neo40 (manufactured by Neogenesis Lab). FIG. 4 (a) is a graph showing the amount of nitrogen oxide released over time ) Is a graph showing cumulative total amount of nitrogen oxides emitted over time.

The amount of NO produced during 12 hours of garlic fermentation was measured for up to 12 hours and no longer thereafter. Neo40 showed up to 2 hours of NO in the same conditions as the fermented garlic, The amount of NO generated during 2 hours was measured because NO did not occur.

From FIG. 3 and FIG. 4, it can be seen that the garlic fermented composition according to the present invention produces nitrogen oxide in an amount of 2500 to 12 times greater than that of the nitrogen oxide supplement.

Until now, there is no example in which R-NO, a precursor of nitric oxide, is immobilized using natural materials. Organic nitrogen (Org-N) is usually nitrified with nitrite nitrogen (NO ₂ ^- N) and instant nitrate nitrogen (NO ₃ ^- N) (FIG. Nitrous nitrogen is not readily immobilized.

The present invention relates to a technique for stably converting and immobilizing nitrite nitrogen (NO ₂ ^- ) in fermented products into R-NO and RS-NO by decomposing organic substances of nitrite nitrogen, which is a precursor of nitric oxide, to be.

R-NO and R-S-NO in the fermentation product are converted to NO in addition to gastric juice or vascular endothelial cells or vitamin C, or in acidic conditions. Existing nitric oxide products are soon converted to NO and their decay time is short. As shown in Experimental Example 2, the natural fermented composition according to the present invention has 12 times more nitrogen oxide than conventional products and is stored in the blood and released gradually.

The present invention relates to a nitric oxide immobilization technique based on a natural substance capable of sufficiently supplementing insufficient nitrogen oxides in the body by using microorganisms and then discharging the remaining nitric oxide to maintain normal nitric oxide balance in the human body.

 The stable R-NO and R-S-NO compounds were constructed by controlling C / N ratio and DO (dissolved oxygen) ratio during fermentation of materials with various high concentration nitrogen compounds.

Nitrogen compounds only take place slowly in the reaction of organic nitrogen (Org-N) to the ammonium nitrogen (NH ₃ -N ^+), The Chemistry nitrite nitrogen (NO ₂ ^- -N) form stable nitrate-nitrogen (NO ₃ ^{- -} N), to rapidly reverse the reaction. The nitrite nitrogen form (NO ₂ ^- N) is converted to nitrate nitrogen (NO ₃ ^- N) in a short time due to unstable binding. Therefore, it is difficult to maintain the form of nitrite nitrogen (NO ₂ ^- N) in the natural world. In addition, even if nitrite nitrogen (NO ₂ ^- N) do.

Accordingly, the present invention was able to convert stable nitric acid into nitrite by regulating the amount of oxygen of Bacillus bacteria, followed by stable R-NO and R-S-NO compounds.

This stable R-NO, R-S-NO compound binds to the red blood cells of the endothelial cells and is released into NO under acidic and anoxic conditions to relax the blood vessels.

TECHNICAL FIELD The present invention relates to a technology for immobilizing nitrogen oxide by constituting stable R-NO and RS-NO compounds by fermenting various nitrogen-containing substances, thereby quantifying and supplying nitrogen oxides regardless of the resistance of vascular endothelial cells caused by aging and diseases to be.

Normally, nitrate nitrogen (NO ₃ ^- N), nitrite nitrogen (NO ₂ ^- N) and NO are called nitric oxide, or antioxidant. Nitrous nitrogen (NO ₂ ^- N) and nitrate nitrogen (NO ₃ ^- N) are precursors of NO and little or no conversion of NO.

In the present invention, a sufficient amount of nitric oxide is converted and fixed from nitrite nitrogen (NO ₂ ^- N) to R - NO and RS - NO compounds to sufficiently fill the deficient amount of nitrogen oxides in the body and the remaining nitric acid is discharged into nitrate nitrogen There are no features.

R-NO and R-S-NO compounds are combined with antioxidant nutrients contained in natural substances to remove oxidative stress (R-O-S) in the body.

Chemicals Sodium nitrite also contains R-NO compounds, but does not relax the hardened blood vessels. In addition, there is a disadvantage in that relaxed blood vessels can not contract. Also, it does not contain any antioxidant substances possessed by natural substances. Therefore, there are side effects such as headache, vomiting, extreme blood pressure, and increased heart rate.

 The in vivo imitation technique according to the present invention is similar to the nitric oxide (NO) produced in the human body, and has the characteristic that the blood vessels are sufficiently relaxed and the remaining nitrogen oxide is released regardless of the hardening of the endothelial cells. The method of producing stable R-NO and RS-NO compounds also solved this problem of ammonia gas in the fermented product, and it can be circulated without preservatives after fermentation.

Unless the organic matter remaining in the nitrite is completely decomposed, the bound R-NO and R-S-NO are converted into NO 3 over time and NO is lost. The present invention is a technology for completely decomposing organic nitrogen to fix nitrogen oxide in the fermented product.

Claims (7)

After mixing the nitrogen-containing organic substance present in the natural world with water and the fermentation strain, the mixture is fermented at room temperature for 5 to 30 days to decompose the sugar and protein of the organic substance and increase the distribution ratio of the bacteria. ;
Converting the carbon source and the oxygen supply to the fermentation product to convert the ammonia nitrogen of the filtrate into nitrite nitrogen and decomposing the remaining organic substances of the nitrite nitrogen and converting and fixing the R-NO and RS-NO to the nitrite nitrogen Of the natural fermented composition.
The method according to claim 1, wherein the nitrogen-containing organic material is at least one of agricultural products, livestock products, forest products, and aquatic products.
The natural fermentation composition according to claim 1, wherein the amount of water is 1 to 20 times (weight ratio) of the nitrogen-containing organic material and the amount of the fermenting bacteria is 0.01 to 1 part by weight based on 100 parts by weight of the mixture of the nitrogen- &Lt; / RTI &gt;
The method of claim 1, wherein the fermenting bacteria is Bacillus subtilis subsp. Subtilis (microorganism deposit number KACC 91554P).
The method according to claim 1, wherein the nitrite nitrogen is converted and immobilized in nitrite nitrogen, RS-NO, and then allowed to stand in an anoxic state for 24 to 36 hours to stabilize the immobilized nitric oxide, &Lt; / RTI &gt; further comprising the steps of:
The method according to claim 1, further comprising an ultrafiltration step of removing nitrifying bacteria and residues in the fermentation product after nitrite nitrogen is converted and immobilized to R-NO and RS-NO, or a sterilization or sterilization step &Lt; / RTI &gt; further comprising the steps of:
7. A natural fermentation composition as claimed in any one of claims 1 to 6.

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