Method for producing low molecular hyaluronic acid

Abstract

To produce a low molecular hyaluronic acid by a simple method.SOLUTION: A method for producing a low molecular hyaluronic acid which decomposes a hyaluronic acid contained in an animal tissue by a splitting enzyme from which Koji is produced by fermentation and produces a low molecular hyaluronic acid, includes: an animal tissue fermentation step of adding Koji to the animal tissue, thereby fermenting the animal tissue, and producing a fermentation decomposition liquid containing at least a low molecular hyaluronic acid. Thereby, a low molecular hyaluronic acid containing a component having a high cosmetic effect by fermentation at low cost can be produced. Low molecular chondroitin can be produced by the animal tissue fermentation step.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a method for producing low molecular weight hyaluronic acid by decomposing animal tissue through fermentation. [Background technology]

[0002] Hyaluronic acid is a type of mucopolysaccharide, a polymeric compound with a molecular weight of several million, consisting of a linear chain of glucosamine and glucuronic acid. It is widely found in animal tissues and has high water retention, making it widely used in the fields of cosmetics, food, medicine, and other industries. However, although high molecular weight hyaluronic acid has high water retention, it has low absorbability and permeability into the body, so there is a demand for lower molecular weight hyaluronic acid. In particular, low molecular weight hyaluronic acid obtained by decomposing animal tissues such as eggshell membrane and chicken comb is said to be effective in promoting hair growth and health promotion due to its penetrating and water-absorbing properties.

[0003] Patent Document 1 describes a method for producing hydrolyzable eggshell membranes by hydrolyzing eggshell membrane powder in an alkaline aqueous solution. [Prior art documents] [Patent documents]

[0004] Patent Document 1: Patent No. 6410229 Summary of the Invention [Problem to be solved by the invention]

[0005] However, the hydrolysis method described in Patent Document 1 has problems such as excessive decomposition of the mucopolysaccharides (hyaluronic acid, chondroitin, etc.) that are contained in the eggshell membrane at about 6% and resulting in the loss of the properties of hyaluronic acid and chondroitin, as well as the need for post-treatment due to the use of chemicals.

[0006] An object of the present invention is to solve the above problems and to produce low molecular weight hyaluronic acid and low molecular weight chondroitin by a simple method. [Means for solving the problem]

[0007] In order to solve the above problems, the present invention provides a method for producing low molecular weight hyaluronic acid by decomposing hyaluronic acid contained in animal tissues with a decomposing enzyme produced by fermentation of koji, comprising the steps of: The present invention provides a method for producing low molecular weight hyaluronic acid, which is characterized by including an animal tissue fermentation step in which koji is added to the animal tissue to ferment it, thereby producing a fermented hydrolyzate solution containing at least low molecular weight hyaluronic acid.

[0008] This composition allows the production of low molecular weight hyaluronic acid containing many components with high beauty effects at low cost through the amylase produced by koji and other various types of enzymatic decomposition. In particular, low molecular weight hyaluronic acid is a component that is easily absorbed by human skin, enhances the skin's moisture retention, and is expected to improve wrinkles caused by dryness. The low molecular weight hyaluronic acid produced here is estimated to have a molecular weight of 10,000 or less.

[0009] In the method for producing low molecular weight hyaluronic acid, the animal tissue fermentation step may be configured to use eggshell membrane for fermentation.

[0010] This configuration makes it possible to produce low molecular weight hyaluronic acid by fermentation using eggshell membranes, which are relatively easy to obtain.

[0011] The method for producing low molecular weight hyaluronic acid may be configured such that the fermentation decomposition liquid produced in the animal tissue fermentation process contains low molecular weight chondroitin produced by decomposing chondroitin contained in the animal tissue.

[0012] This configuration makes it possible to produce, in addition to low molecular weight hyaluronic acid, low molecular weight chondroitin that is easily absorbed by human skin.

[0013] In the method for producing low molecular weight hyaluronic acid, the animal tissue fermentation step may be configured to use chicken comb for fermentation.

[0014] This configuration makes it possible to produce low molecular weight hyaluronic acid using rooster comb, which has not been used much until now.

[0015] The method for producing low molecular weight hyaluronic acid may further include a koji production step of adding the animal tissue to seed koji to produce the koji prior to the animal tissue fermentation step.

[0016] With this configuration, by culturing koji mold from the seed koji, the koji mold can produce protein decomposition enzymes such as protease and sugar decomposition enzymes such as amylase.

[0017] The method for producing low molecular weight hyaluronic acid may further include a step of sterilizing the fermented decomposition liquid and inactivating the enzymes therein after the animal tissue fermentation step.

[0018] This configuration stops the excessive decomposition of proteins and mucopolysaccharides by the proteases and amylases contained in the filtered hydrolyzed liquid, and stops the growth of koji mold.

[0019] The method for producing low molecular weight hyaluronic acid may further include a solid matter removal step of removing solid matter from the fermentation decomposition liquid after the inactivation step.

[0020] This configuration makes it possible to remove undecomposed eggshell membranes and koji mold cells, which are solid contents that are not required as cosmetic raw materials.

[0021] The method for producing low molecular weight hyaluronic acid may further include a drying step of drying the fermentation decomposition liquid after the solid content has been removed in the solid content removal step.

[0022] This composition makes it possible to obtain a powder of low molecular weight hyaluronic acid that is easy to use. Effect of the Invention

[0023] According to the method for producing low molecular weight hyaluronic acid of the present invention, low molecular weight hyaluronic acid having high cosmetic effects can be produced at low cost. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS EXAMPLES

[0024] The method for producing low molecular weight hyaluronic acid in Example 1 of the present invention will be described. The method for producing low molecular weight hyaluronic acid in Example 1 includes an animal tissue fermentation step in which hyaluronic acid contained in animal tissue is decomposed by protease such as protease produced from koji.

[0025] The applicant previously succeeded in producing fermented decomposition eggshell membrane liquid by fermenting eggshell membrane (Patent No. 7237291, Patent No. 7237292), and this time, he further discovered that the fermented decomposition liquid contains hyaluronic acid that has been decomposed into low molecular weight by fermentation. Low molecular weight hyaluronic acid is an ingredient that is easily absorbed by the human body, increases the skin's moisturizing properties, and is expected to improve wrinkles caused by dryness.

[0026] In Example 1, an animal tissue fermentation process is carried out to produce a fermentation decomposition liquid containing at least low molecular weight hyaluronic acid. In the animal tissue fermentation process, coarsely crushed eggshell membrane is placed in a tank and koji is added. Any koji such as soy sauce koji, rice koji, or soybean koji can be used as the koji in Example 1. When koji is added to 10 to 100 g of eggshell membrane, enzymatic decomposition occurs at approximately 25°C to 45°C. When the mixture is matured for 0.5 to 7 days in this state, proteins and mucopolysaccharides are decomposed by the protease enzyme, peptidase enzyme, and amylase secreted by the koji. When fermentation is completed, a turbid liquid containing at least low molecular weight hyaluronic acid (hereinafter also referred to as "fermentation decomposition liquid") is obtained.

[0027] As a result of the fermentation decomposition by the animal tissue fermentation process, the molecules of hyaluronic acid before fermentation are decomposed, and a fermentation decomposition liquid containing low molecular weight hyaluronic acid is obtained. It was also found that the fermentation decomposition liquid contains low molecular weight chondroitin. Like low molecular weight hyaluronic acid, low molecular weight chondroitin is easily absorbed by human skin, increasing the skin's moisture retention and improving wrinkles caused by dryness.

[0028] In Example 1, eggshell membrane was used as the animal tissue, but it is not limited thereto and can be appropriately changed. For example, low molecular weight hyaluronic acid and low molecular weight chondroitin can be produced by carrying out the animal tissue fermentation process using chicken comb.

[0029] Thus, in Example 1, a method for producing low molecular weight hyaluronic acid is provided in which hyaluronic acid contained in animal tissue is decomposed by the decomposition enzymes produced by fermentation of koji, and low molecular weight hyaluronic acid is produced. The method for producing low molecular weight hyaluronic acid is characterized by including an animal tissue fermentation step in which koji is added to the animal tissue to ferment it and produce a fermented hydrolyzate solution containing at least low molecular weight hyaluronic acid. This makes it possible to produce low molecular weight hyaluronic acid and low molecular weight chondroitin, which have high cosmetic effects, at low cost in a simple manner. EXAMPLES

[0030] Example 2 of the present invention is different from Example 1 in that a koji production step of producing koji from seed koji is provided prior to the animal tissue fermentation step.

[0031] In the koji production process in Example 2, koji is produced from seed koji. 10-100 g (dry weight) of eggshell membranes, which have also been sterilized, and 10-100 g of glucose are added to 5 L of sterilized water, and 10 ml of koji mold liquid is added, followed by stirring with aeration at 25°C to 45°C for one day to one week. During this time, the koji mold produces protein-degrading enzymes such as protease and peptidase, and amylase, a sugar-degrading enzyme. These enzymes are then dissolved into the liquid.

[0032] Next, the koji produced in the koji production process is used to carry out an animal tissue fermentation process to produce a fermentation decomposition liquid containing at least low molecular weight hyaluronic acid. In the animal tissue fermentation process, roughly crushed eggshell membrane or cockscomb is placed in a tank, and the koji produced in the koji production process is added. The koji in the second embodiment can be any koji, such as soy sauce koji, rice koji, or soybean koji. When koji is added to 10 to 100 g of eggshell membrane, enzymatic decomposition occurs at about 45°C. When the mixture is matured for 0.5 to 7 days in this state, proteins and sugars are decomposed by the protease enzyme, peptidase enzyme, and amylase secreted by the koji. When the fermentation is completed, a turbid liquid containing low molecular weight hyaluronic acid (hereinafter also referred to as "fermentation decomposition liquid") is produced.

[0033] In this way, in Example 2, a koji production step of producing koji from seed koji is provided prior to the animal tissue fermentation step. By cultivating koji from seed koji, the koji mold can produce protease and other protein-degrading enzymes and amylase and other sugar-degrading enzymes. EXAMPLES

[0034] Example 3 of the present invention is a method for producing low molecular weight hyaluronic acid, and is different from Example 1 and Example 2 in that it includes a process for sterilization and inactivation of enzymes in the fermentation decomposition liquid after the animal tissue fermentation process.

[0035] In Example 3, after the animal tissue fermentation process, an inactivation process is carried out to sterilize and inactivate the enzymes. As a result, the fermentation decomposition liquid contains low molecular weight hyaluronic acid and low molecular weight chondroitin even after filtration, but by carrying out the inactivation process, further decomposition of peptides is prevented, the proliferation of koji mold is stopped, and spoilage due to various bacteria can be prevented.

[0036] In the inactivation process, the fermented and decomposed eggshell membrane liquid that has been decomposed in the animal tissue fermentation process may be heated at 60 to 90°C for 10 minutes to 3 hours, but in many cases, sterilization and enzyme inactivation can be achieved by heating at 70°C for 30 minutes.

[0037] Thus, in the third embodiment, the method for producing low molecular weight hyaluronic acid includes a process for sterilizing the fermented decomposition liquid and inactivating the enzymes therein after the animal tissue fermentation process, which prevents the proteases contained in the filtered decomposition liquid from decomposing the koji mold and other bacteria, and prevents the bacteria from multiplying and causing spoilage. EXAMPLES

[0038] Example 4 of the present invention is different from Example 3 in that it includes a solids removal step after the inactivation step, in which solids are removed from the inactivated fermentation decomposition liquid to obtain a fermentation decomposition liquid.

[0039] In Example 4, after the inactivation step, a solid content removal step is carried out to obtain a fermented decomposition liquid by removing solid content from the inactivated fermented decomposition liquid, which makes it possible to remove undecomposed eggshell membranes, cockscomb, and Aspergillus oryzae cells, which are solid content not required as cosmetic raw materials.

[0040] Specifically, the solid content removal step can be carried out by placing the fermentation-decomposed eggshell membrane liquid in a cloth bag or the like and squeezing it. Alternatively, the solid content may be removed by filtration using filter paper or the like, or by using a centrifuge.

[0041] Thus, in Example 4, the method for producing low molecular weight hyaluronic acid includes a solid matter removal step for removing solid matter from the fermentation decomposition liquid after the inactivation step, which makes it possible to remove undecomposed eggshell membranes and koji mold cells, which are solid matters not required as cosmetic raw materials. EXAMPLES

[0042] Example 5 of the present invention is different from Example 4 in that it includes a drying step of drying the fermentation decomposition liquid after the solid content has been removed in the solid content removing step.

[0043] In Example 5, a drying step is carried out to dry the fermentation decomposition liquid after the solid content has been removed by the solid content removal step. The drying step can be carried out by a known method such as a spray drying method. The drying step can produce a powder of fermentation decomposition eggshell membranes that is easy to use.

[0044] Thus, in Example 5, a method for producing low molecular weight hyaluronic acid is provided that includes a drying step for drying the fermentation decomposition liquid after the solids have been removed in the solid removal step, thereby making it possible to obtain fermentation decomposition eggshell membrane in a powder form that is easy to use. [Industrial Applicability]

[0045] The method for producing low molecular weight hyaluronic acid in the present invention is not limited to eggshell membranes and chicken combs, but can be widely used in fields that utilize fermentation decomposition of animal tissues such as dairy products and egg yolks.

Claims

1. A method for producing low molecular weight hyaluronic acid by decomposing hyaluronic acid contained in animal tissues with a decomposition enzyme produced by fermentation of koji, comprising the steps of: The method for producing low molecular weight hyaluronic acid is characterized by including an animal tissue fermentation step in which koji is added to the animal tissue to ferment it, thereby producing a fermented decomposition liquid containing at least low molecular weight hyaluronic acid.

2. 2. The method for producing low molecular weight hyaluronic acid according to claim 1, wherein the animal tissue fermentation step uses eggshell membrane for fermentation.

3. The method for producing low molecular weight hyaluronic acid according to claim 2, characterized in that the fermentation decomposition liquid produced in the animal tissue fermentation process contains low molecular weight chondroitin obtained by decomposing chondroitin contained in the animal tissue.

4. 2. The method for producing low molecular weight hyaluronic acid according to claim 1, wherein the animal tissue fermentation step uses chicken comb for fermentation.

5. 2. The method for producing low molecular weight hyaluronic acid according to claim 1, further comprising a koji production step of adding the animal tissue to seed koji to produce the koji prior to the animal tissue fermentation step.

6. The method for producing low molecular weight hyaluronic acid according to any one of claims 1 to 5, characterized in that the method further comprises a process for sterilizing and inactivating the enzyme in the fermentation decomposition liquid after the animal tissue fermentation process.

7. The method for producing low molecular weight hyaluronic acid according to claim 6, further comprising a solid matter removal step of removing solid matter from the fermentation decomposition liquid after the inactivation step.

8. The method for producing low molecular weight hyaluronic acid according to claim 7, further comprising a drying step of drying the fermentation decomposition liquid after the solid content has been removed by the solid content removal step.