JP2019208495A - Ergothioneine-containing composition - Google Patents

Abstract

To provide compositions containing ergothioneine, to provide foods using the same, and additionally to provide effective uses of the mycelial residue of Pycnoporus basidiomycete used in enzyme production.SOLUTION: The hot-water extract of Pycnoporus basidiomycete, desirably Pycnoporus coccineus basidiomycete, contains ergothioneine. In addition, since it also contains polyphenol and free amino acid, it can be used as a food material. As the mycelium of Pycnoporus basidiomycete, a residue produced as a by-product when the bacterium produces various enzymes can also be used.SELECTED DRAWING: None

Description

The present invention relates to an extract of the genus Pycnoporus basidiomycetes containing ergothioneine which is an antioxidant.

Ergothioneine is one of the hydrophilic sulfur-containing amino acids and is an antioxidant useful for the human body.
Ergothioneine can be biosynthesized by fungi and mycobacteria, which are absorbed by the plant from the roots, and animals ingest ergothioneine by eating the plant. For this reason, it has been difficult for humans to take a sufficient amount of ergothioneine for the human body through a normal diet.

 In order to ingest ergothioneine efficiently, a method for culturing basidiomycetes and extracting ergothioneine therefrom has been reported (Patent Document 1).

On the other hand, since the culture filtrate and extract of basidiomycetes belonging to the genus Pycnoporus contain various enzymes, the production of yeast extract (Patent Document 2), the enzyme-degraded seasoning of animal proteins (Patent Document 3), plant proteins It is known that it is suitably used for the production of food and drink, such as the enzyme-decomposable seasoning (Patent Document 4), the extraction of tea (Patent Document 5), the production of fish salmon (Patent Document 6), and the like.

In addition, oyster mushroom (Pycnoporus coccineus), one of basidiomycetes, is used as a raw material for glucanase, cellulase, protease, hemicellulase, and polyphenol oxidase, which are enzymes for food production. (Non-Patent Document 1)
After cultivating the bamboo shoot and extracting the enzyme, the mycelium became a residue and had no use.

JP 2011-102286 A, International Publication WO 2014/148132 A, JP 2014-223051 A, JP 2015-116173 A JP 2004-248529 A, JP 2006-129835 A JP 2006-094756 A JP 2006-094757 A JP 2008-125477 A JP 2008-212116 A

Official Food Addendum 9th Edition

An object of the present invention is to provide a composition containing ergothioneine and a food using the same.
Furthermore, it is effective utilization of the residue produced when an enzyme is produced by culturing basidiomycetes belonging to the genus Pycnoporus.

The present inventors have found that an ergothioneine-containing composition can be obtained by extracting the mycelium-containing residue after obtaining the enzyme from the culture solution of Pycnoporus genus basidiomycetes with hot water.
In addition to ergothioneine, the hot water extract contains polyphenols and a tasteful amino acid, so it has functions of maintaining color tone, suppressing off-flavor, and imparting flavor, and is suitable as a food material. Found to be used.

That is, the present invention relates to the following (1) to (5).
(1) Mycelium of Pycnoporus basidiomycetes containing 0.5 mg / g or more of ergothioneine per dry weight and 2 mg / g or more of polyphenol per dry weight.
(2) A Pycnoporus basidiomycetous extract containing 1 mg / g or more of ergothioneine per dry weight.
(3) A Pycnoporus basidiomycetous extract containing 1 mg / g or more of ergothioneine per dry weight and 10 mg / g or more of polyphenol per dry weight.
(4) The extract according to (2) or (3) above, wherein the Pycnoporus genus basidiomycete is Pycnoporus coccineus.
(5) A food material containing the Pycnoporus genus basidiomycetous extract according to any one of (2) to (4) above.

According to the present invention, an ergothioneine-containing composition can be obtained by a simple process of culturing Pycnoporus basidiomycetes and extracting from mycelium. The composition can be used as a raw material for ergothioneine high-purity products.
In addition to ergothioneine, the composition extracted from Pycnoporus basidiomycetes also contains polyphenols and tasteful amino acids, and has the functions of color retention, off-flavor suppression, and imparting flavor. It can be suitably used as a food material.
Furthermore, since this ergothioneine-containing composition can be extracted from the mycelium-containing residue of the oyster mushroom used for obtaining the enzymes, it is possible to obtain the residue effectively and at a low cost.

The extract of the present invention is obtained from a basidiomycete belonging to the genus Pycnoporus, which is one of the basidiomycetes.
The basidiomycetes belonging to the genus Pycnoporus include Pycnoporus coccineus (Pleurotus), Pycnoporus cinnabarinus (Pleurotus), Pycnoporus palibini, Pycnoporus puniceus, Pycnoporus sanguineus, etc. Among them, Pycnoporus coy

Culture of basidiomycetes belonging to the genus Pycnoporus may be performed by a known method.
For example, a spore suspension of Pycnoporus basidiomycetes is inoculated into a liquid medium and cultured. The culture may be divided into two stages: seed culture and main culture.

Examples of the components of the liquid medium include calcium chloride, magnesium sulfate, ammonium sulfate, monopotassium phosphate, sucrose, corn steep liquor, defatted soybean flour, dextrin, thiamine hydrochloride, and the like.

After inoculating the prepared liquid medium with a spore suspension of Pycnoporus genus basidiomycetes, the mycelium grows by, for example, stirring culture at 28-30 ° C. for 2-6 days, and at the same time, an enzyme useful for food production Kinds are also produced.

A solid content containing Pycnoporus basidiomycetous mycelium is separated from the culture broth after culturing. Separation methods include filtration using a filter paper, membrane, diatomaceous earth, etc., and centrifugation. The obtained mycelium-containing material may be washed with a solvent such as ethanol. The culture solution after the mycelium separation contains useful enzymes such as glucanase, cellulase, protease, hemicellulase, polyphenol oxidase, etc., and can be used as an enzyme composition as it is or after purification.

The other Pycnoporus basidiomycete mycelium contains ergothioneine and polyphenol, so that the mycelium itself or mycelium extract can be used effectively. Although the ergothioneine content in the mycelium varies depending on the strain and culture conditions, it is generally about 0.5 mg / g, more preferably 0.8 mg / g or more per dry mycelium. The polyphenol content in the mycelium varies depending on the strain, culture conditions, etc., but is generally about 2 mg / g, more preferably 4 mg / g or more per dry mycelium.

The mycelium or its contents are freeze-dried or mechanically pulverized and then immersed in a solvent to extract the extract. The solvent is good as a food, and water, a mixture of alcohol and water are particularly desirable. When water is used as the extraction solvent, the temperature is desirably 80 to 100 ° C., and the immersion time is desirably 20 to 120 minutes. The suspension after the immersion is separated into an extract residue and an extract solution of basidiomycete mycelium by centrifugation or filtration.

The Pycnoporus basidiomycetous extract solution thus obtained contains ergothioneine. This solution may be concentrated and dried to obtain a dry powder of an extract from the genus Pycnoporus.
The ergothioneine content in the Pycnoporus genus basidiomycete extract varies depending on the strain, culture conditions, and extraction conditions, but is approximately 1 mg / g or more in the dry matter.

The obtained Pycnoporus genus basidiomycete extract may be used as a raw material for obtaining high-purity ergothioneine. When used as a raw material for high-purity ergothioneine, the ergothioneine content in the extract should be high, desirably 2.0 mg / g or more per dry weight, more desirably 3.0 mg / g or more.

The Pycnoporus genus basidiomycetous extract contains 10 mg / g or more of polyphenol per dry weight in addition to ergothioneine. Furthermore, it contains various components such as free amino acids, some of which have taste. And, when added to food, it has the effect of suppressing off-flavor of food or preventing discoloration, and depending on the amount added, it can be given flavor, so it can be suitably used as a food material it can.

<Quantitative determination of ergothioneine>
A test sample obtained by diluting the Pycnoporus basidiomycetous extract obtained by the above method to an appropriate concentration with water is used.
Use a HILIC column for HPLC, and use a mixture of 10 mM ammonium acetate and acetonitrile 1: 4 as the mobile phase. The flow rate is 0.5 mL / min, the column temperature is 40 ° C., and a peak derived from ergothioneine is detected by absorption at 220 nm with a UV detector.
Separately, the peak area of the ergothioneine preparation of a predetermined concentration is measured by HPLC, and the ergothioneine amount of the test sample is calculated by comparing the areas of the peaks appearing at the same time.

<Quantification of polyphenol>
Using a Pycnoporus basidiomycete extract as a sample, the polyphenol content is quantified by the Folin-Denis method.

The Pycnoporus genus basidiomycetous extract of the present invention contains ergothioneine and polyphenol, both of which are substances having an antioxidant function, and can be used as a material for functional foods. Moreover, by adding the Pycnoporus genus basidiomycetes extract of the present invention to a food, mixing, surface treatment, etc., there is an effect of suppressing off-flavor of the food or preventing discoloration.

Foods that are expected to prevent discoloration mainly include livestock raw materials (meat such as beef, pork and chicken), fishery materials (fish such as tuna and yellowtail, shellfish such as shrimp and crab), fruits, etc. (Cut fruits such as apples, bananas and melons, pickled syrup, etc.), vegetables (cut vegetables such as lettuce and cabbage, pickled eggplants, potatoes, lotus root, avocado, etc.), processed grains (noodles, bread, cooked rice) ), Processed fats and oils (mayonnaise, margarine, etc.), processed meat (ham, sausage, etc.), processed fishery products (kamaboko, chikuwa, fried fish cakes, etc.), dairy products (butter, cheese, yogurt, etc.), fruit processing Goods (jam, marmalade, etc.), confectionery (chocolate, cookies, cakes, jelly, etc.), various beverages (juice, coffee, tea, green tea, carbonated drinks, etc.) , Seasoning (soy sauce, source, mirin, etc.), and the variety of food.

Examples of foods that are expected to have an off-flavor suppression effect include livestock products (raw materials such as beef, pork and chicken, processed products), and marine products (raw fish, tsumire).

As a method for adding the Pycnoporus genus basidiomycetous extract of the present invention to food, there are methods such as mixing into food, spraying on the surface, application, and immersion.
When mixing with food for the main purpose of preventing discoloration, it is desirable that the ergothioneine concentration per food be 50 ppm or more. When surface treatment such as spraying, coating, or dipping is performed, spray, coating, or dipping It is desirable that the ergothioneine concentration in the liquid be 50 ppm or more.
When the main purpose is to control off-flavor, it is desirable to add polyphenol content per food to 30 ppm or more.

EXAMPLES Hereinafter, although an Example demonstrates this invention, this invention is not limited to these Examples.

<Example 1>
20 mL of spore suspension of Pycnoporus coccineus (10 ⁷ cells / mL or more) in seed medium (calcium chloride 1 g / L, magnesium sulfate 1 g / L, ammonium sulfate 2 g / L, monopotassium phosphate 2 g / L, sucrose 50 g / L, corn steep liquor 30 g / L, pH 7.0) was inoculated into 200 mL, and cultured in a 2 L flask at 28 ° C. and 200 rpm for 48 hours to obtain a seed culture end solution.

200 mL of the obtained seed culture end solution was added to the main medium (calcium chloride 1 g / L, magnesium sulfate 1 g / L, ammonium sulfate 2 g / L, monopotassium phosphate 2 g / L, sucrose 80 g / L, defatted soybean flour 35 g / L, The cells were transplanted to 2 L of pH 6.0) and cultured in a 20 L jar at 28 ° C. and 200 rpm for 96 hours to obtain a main culture end solution. The main culture end liquid was centrifuged to separate into a filtrate having enzyme activity and a mycelium-containing material.

1000 g of the obtained mycelium-containing material was washed with ethanol and lyophilized to obtain 200 g of a dried mycelium-containing material. This was suspended in 750 mL of hot water and stirred at 90 ° C. for 30 minutes. The suspension after the hot water treatment was separated into a mycelium extraction residue of basidiomycetes and an extract solution by centrifugation. Ergothioneine contained 1.0 mg / g per residue containing dry mycelium and polyphenols contained 5.4 mg / g per residue containing dry mycelium.

The resulting Hiyotake extract solution was concentrated 5 times to obtain a concentrated solution (Sample 1) of Hiirotake extract solution having a solid content concentration of 37.4% by weight. Sample 1 contained 1,299 ppm ergothioneine, 7,149 ppm polyphenol, and also contained free amino acids.
Therefore, the obtained bamboo shoot extract contained 3.5 mg / g ergothioneine and 19.1 mg / g polyphenol per dry weight.

<Example 2>
A cooking test was conducted with the concentrated solution of the bamboo shoot extract solution obtained in Example 1 (Sample 1).

-The sliced eggplant was soaked for 1 hour and a half in a soaked seasoning solution to which 5% of soaked sample 1 was added. The liquid was then drained and stored refrigerated for 1 day. About the eggplant after a preservation | save, the color tone was compared by making the thing immersed similarly in the shallow pickle seasoning liquid which did not add the sample 1 as a control | contrast. The eggplant of the control turned brown, whereas the eggplant soaked in the seasoning liquid added with Sample 1 had no discoloration and maintained the state at the time of cutting.

-Beef sample 1 was applied to the surface of the meat corresponding to 5% by weight and stored at 10 ° C for 4 days. About the color tone of the beef after a preservation | save, the color tone was compared by making the thing which did not apply | coat the sample 1 into a control | contrast. The control beef turned brown, while the beef coated with Sample 1 maintained a bright red color.

-1% of sample 1 was added to and mixed with hamburger seeds before baking. Thereafter, it was stored at 10 ° C. for 1 day, and the amount of hexanal, which is the main causative substance of the oxidized odor of fat, was measured and its flavor was evaluated. As a result, when the sample 1 was not added as a control, the amount of hexanal generated was reduced, and a reduction in meat odor and umami were imparted.

As described above, the Pycnoporus genus basidiomycete extract of the present invention contains ergothioneine, and therefore can be used as a food material having an antioxidant function, and also as a raw material for high-purity ergothioneine.
Moreover, since it has the color tone retention effect, the off-flavor suppression effect, the umami imparting effect, etc. with respect to the added food, it can be used suitably as a food material.

Claims (5)

Mycelium of Pycnoporus genus basidiomycetes containing ergothioneine 0.5 mg / g or more per dry weight and polyphenol 2 mg / g or more per dry weight. An extract of the genus Pycnoporus basidiomycetes containing 1 mg / g or more of ergothioneine per dry weight. An extract of the genus Pycnoporus basidiomycetes containing 1 mg / g or more of ergothioneine per dry weight and 10 mg / g or more of polyphenol per dry weight. The extract according to claim 2 or 3, wherein the basidiomycete belonging to the genus Pycnoporus is Pycnoporus coccineus. The food material containing the Pycnoporus genus basidiomycetes extract in any one of Claims 2-4.