TWI827804B - Methods for producing mushroom mycelium, compositions containing mushroom mycelium, and food materials using mushroom mycelium - Google Patents

Abstract

Various plant-based materials are being studied as meat substitutes. However, when soybeans are used, they may have a grainy taste unique to soybeans, which may impair the flavor and may become an allergen for food allergies. When the fruiting bodies of mushrooms are used as meat substitutes, it is possible to become a meat substitute material that has the same taste as processed meat products. However, when mushroom mycelium is used, low productivity is a big problem. In addition, when using mushroom mycelium as a meat substitute, there are issues such as taste and flavor. In order to solve the above-mentioned problems, the inventors of the present invention discovered that by using mushroom mycelium and using specific culture medium components, the productivity of the mushroom mycelium can be improved, and thus completed the present invention.

Description

Methods for producing mushroom mycelium, compositions containing mushroom mycelium, and uses Food materials using mushroom mycelium

The present invention relates to the use of mushroom mycelium as a meat substitute material and meat-like foods made using the material.

In recent years, health awareness has increased, and foods consumed every day are required to be low in calories and fat. Meat is a necessary food because it is rich in nutrients such as protein. However, it also contains fat. Therefore, excessive intake may cause various chronic diseases including metabolic syndrome.

Furthermore, as the world's population increases, it is conceivable that not only meat sources, but also animal protein sources such as fish will become scarce and prices will rise.

In this way, meat substitute materials that can be used as a source of protein in addition to edible meat are being developed.

As far as meat substitutes go, some use plant-based proteins. Among plants, soybean-based proteins are rich in protein, and various proposals have been made as a meat substitute (Patent Documents 1 and 2).

As plant-based materials other than soybeans, mushrooms are used (Patent Documents 3 and 4). Regarding mushrooms, the use of mycelium and fruiting bodies has been studied (Patent Document 5).

However, in order to use mushroom mycelium as a meat substitute material, a large amount of culture is required. The production period of mycelium is generally shorter than that of fruiting bodies of mushrooms, but in the case of liquid culture, the growth rate is slow, the culture time is long, and the productivity tends to be low (Patent Documents 3 and 5). Patent Document 3 describes cultivating mushroom mycelium, and a maximum of 24g/L/9Day mycelium can be obtained.

Therefore, it is necessary to cultivate mushroom mycelium with high efficiency so that it can have the taste and nutritional content suitable as a meat substitute material.

On the other hand, γ-aminobutyric acid (GABA) is an amino acid produced by the decarboxylation of glutamic acid in the body and does not form a protein. As a neurotransmitter, it plays an important role in the central nervous system. When ingested, GABA has the effect of suppressing the rise in blood pressure and calming the mind. Therefore, various foods containing GABA are being developed. Mushrooms contain only a small amount of GABA, but enoki mushrooms, shiitake mushrooms, and the like containing high levels of GABA are being developed (Patent Document 6).

[Prior technical literature]

[Patent Document]

[Patent Document 1] Japanese Patent Application Publication No. 10-179098

[Patent Document 2] Japanese Patent Application Publication No. 2012-75358

[Patent Document 3] Japanese Patent Publication No. 2009-538128

[Patent Document 4] Japanese Patent Application Publication No. 2018-50607

[Patent Document 5] Japanese Patent Application Publication No. Sho 64-23874

[Patent Document 6] Japanese Patent Application Publication No. 2008-301798

In this way, various plant-based materials are being studied as meat substitutes. However, when soybeans are used, they may have a grainy smell unique to soybeans, which may impair the flavor and may become an allergen for food allergies. When the fruiting bodies of mushrooms are used as meat substitutes, it is possible to become a meat substitute material that has the same taste as processed meat products. However, when mushroom mycelium is used, low productivity is a big problem. In addition, when using mushroom mycelium as a meat substitute or food, there are issues such as taste and flavor.

In order to solve the above-mentioned problems, the inventors of the present invention discovered that by using mushroom mycelium and using specific culture medium components, the productivity of the mushroom mycelium can be improved, and thus completed the present invention.

The present invention is: (1) A method for producing mushroom mycelium, which includes the following steps: in a culture medium containing γ-aminobutyric acid (GABA), a GABA precursor, or a substance that can generate GABA through metabolism Culture to form mushroom mycelium; (2) A composition containing: cultured in a culture medium containing γ-aminobutyric acid (GABA), a GABA precursor, or a substance that can be metabolized to generate GABA. Mushroom mycelium; (3) A food containing the composition of (2).

According to the present invention, a culture medium containing γ-aminobutyric acid (GABA) or the like is used to improve the productivity of mushroom mycelium, especially Pleurotus eryngii mycelium. In addition, when used as a meat substitute material, according to the present invention The mycelium obtained by this method unexpectedly turns red, so its appearance is similar to meat, and it can be ideally used as a meat substitute material.

In addition, the mushroom mycelium obtained by the method of the present invention has a higher dietary fiber content than mycelium obtained by ordinary culture methods, and therefore can also be used as dietary fiber-fortified food.

The present invention provides a composition for cultivating mushroom mycelium and using it as a meat substitute material and a manufacturing method thereof.

The mushroom mycelium of the present invention refers to an aggregate of hyphae formed by spores budding and repeated cell division. On the other hand, fruiting body refers to the structure formed by fungi from hyphae in order to form spores. It is a structure that is generally used as the edible part of mushrooms and is clearly distinguished from mycelium.

The mushroom mycelium that can be used in the present invention is among the mushrooms, belongs to the phylum Basidiomycetes, and can be used as Basidiomycetes for edible or medicinal purposes. For example, Pleurotus eryngii, Pleurotus cystidiosus, Pleurotus osteatus, Pleurotus pulmonarius, etc. of the family Pleurotus, Schizophyllum commune of the family Schizophyllum. etc., Basidiomycetes such as Flammulina velutipes and Ganoderma lucidum.

The aforementioned basidiomycetes that form the mycelium of Pleurotus eryngii can be used without particular restrictions as long as they are edible. They can be commercially available strains, wild strains, tissue isolates derived from fruiting bodies, etc. that are generally available. .

In the present invention, mycelium obtained by culturing with a general culture medium composition used for culturing basidiomycetes such as Pleurotus eryngii is used. In the present invention, it is preferable to use liquid culture medium for culture. In liquid culture media, carbon sources are generally added, and nitrogen sources are added as needed. Inorganic salts such as magnesium salts and phosphates, and vitamins can also be added.

As the carbon source, those generally used for culturing basidiomycetes can be used, and generally examples include glucose, fructose, lactose, trehalose, starch, and the like. Generally speaking, the amount added to the culture medium is less than 20% by weight relative to the culture medium, and it is advisable to add 1 to 5%. The concentration of the carbon source can be adjusted to maintain a fixed concentration during the culture process, or can be adjusted to the aforementioned concentration only at the beginning of the culture.

As a nitrogen source, yeast extract, polyprotein, casein, ammonium sulfate, ammonium nitrate, etc. can be used. Regarding the added concentration of the nitrogen source, it is advisable to add it so that the total added amount of these components becomes 0.1~3.0%.

In the liquid culture medium, magnesium sulfate, zinc sulfate, and other sulfates, or inorganic salts such as magnesium salts and phosphates, or vitamins, etc. can also be added. As for the inorganic salts, in addition to the above examples, potassium dihydrogen phosphate, sodium chloride, manganese sulfate, iron sulfate, cobalt sulfate, copper sulfate, potassium sulfate, aluminum, boric acid, etc. can also be added.

In the present invention, γ-aminobutyric acid (GABA) is added to the culture medium. The concentration of GABA added to the culture medium can be adjusted appropriately, but is usually 0.01 to 10% by weight in terms of the solid content in the culture medium. It is advisable to add 0.5% to 2% by weight to the culture medium.

In the present invention, the following method can be used: even if GABA is not directly added, GABA can be produced or synthesized in the basidiomycete fungus or the culture medium through metabolism in the basidiomycete fungus or in the culture medium. Carry out cultivation. For example, the method of producing GABA from glutamic acid. Specifically, it may be a method of adding glutamic acid to the culture medium, or simultaneously adding glutamic acid and a substance having glutamic acid decarboxylase (GAD) activity. In addition, by adding polyamines (such as spermine, spermidine, putrescine), basidiomycetes will metabolize and produce GABA, so polyamines can also be added to the culture medium. method.

When using a method of culturing under conditions that allow GABA to be produced or synthesized in basidiomycetes or in a culture medium, for example, when adding polyamines or glutamic acid, the amount to be added can be determined by those with general knowledge in the technical field. The amount can be adjusted appropriately. For example, if it is glutamic acid, 0.1~10% by weight is usually added. For polyamines, add 0.001~1% by weight. In addition, yeast extracts containing polyamines can also be used. At this time, the amount of yeast extract added can be adjusted according to the content of polyamines in the yeast extract.

In addition, other components added to the culture medium are not particularly limited, and organic salts and the like may also be added. As for organic salts, acetic acid, lactic acid, citric acid, malic acid, formic acid, etc. can also be added.

Mix the above ingredients to prepare a medium for forming Pleurotus eryngii mycelium. A general method can be used for a mixing method etc., and there is no particular restriction. It can be adjusted by the usual method of adjusting the culture medium when cultivating basidiomycetes. The method of mixing the culture medium with γ-aminobutyric acid (GABA), GABA precursors, or substances that generate GABA through metabolism is also not particularly limited. It is also possible to mix and culture one or more substances selected from γ-aminobutyric acid (GABA), a GABA precursor, or a substance that generates GABA through metabolism. For example, GABA, polyamines, etc. may be mixed and added.

Basidiomycetes can be cultured according to normal culture conditions. For example, the pH may be within a general range suitable for basidiomycetes. To form Pleurotus eryngii mycelium, it needs to be 3.0~8.0, preferably 4.0~7.0. culture temperature The temperature can be carried out within the temperature range commonly used for cultivating basidiomycetes, preferably 22 to 35°C, more preferably 25 to 32°C. In addition, liquid culture can be carried out under aeration and stirring conditions for 2 to 30 days.

The stirring speed varies depending on the culture scale. A general stirring speed can be used in the culture of basidiomycetes. Usually it is carried out at 150rpm~400rpm.

In addition, in the present invention, culture can also be divided into preculture and main culture generally used in mycelial culture. The main culture is a step of cultivating Pleurotus eryngii mycelium in large quantities, and the pre-culture is usually a step of cultivating it in a smaller capacity than the main culture. In the present invention, pre-culture and main culture can be performed under the same conditions except for the culture capacity. In addition, gamma-aminobutyric acid (GABA), GABA precursors, or substances that generate GABA through metabolism do not need to be added in the preculture.

The mycelium cultured as mentioned above can be used as imitation meat food. The mycelium system is used after the solid components are separated by centrifugal separation, etc. After the solid components are recovered, they can be used directly or dried before use.

According to the culture method of the present invention, when GABA, etc. are added, a yield of 24 to 35 g/L on a dry basis can be obtained. Compared with the yield of 11 to 13 g/L without adding GABA, etc., productivity can be greatly improved. , it is useful in the utilization of food and the utilization of replacing edible meat.

The mycelium of the present invention can be used alone as a meat substitute material, or it can be mixed with root vegetables such as soybeans and okara, cereals such as bread flour, plant-based protein materials such as hydrolyzate of plant-based protein (HVP), eggs, More than one type of protein, etc.

In addition, the meat substitute material is not limited to foods that do not use edible meat, and edible meat and the mycelium of the present invention can also be mixed.

By using the mushroom mycelium of the present invention alone or mixed with the ingredients exemplified in the previous paragraph, it can be substituted for, for example, meat-based foods such as hamburger steaks, meatballs, dumplings, siomai, meat buns, etc. Imitation meat products can be made by using livestock meat used in processed meat products such as stuffing, sausage, ham, and bacon.

In addition, for example, part of the livestock meat in the hamburger steak may be replaced with the Pleurotus eryngii mycelium of the present invention, and part of the livestock meat may be replaced with the product of the present invention. In addition, since the mushroom mycelium obtained by the method of the present invention produces red pigment, it can be made into imitation meat food with an appearance like red meat of livestock meat.

In addition, not only as a meat substitute food, mushroom mycelium such as Pleurotus eryngii mycelium of the present invention contains more protein than the fruiting bodies of mushrooms, so it can also be used as a protein-containing food. In addition, the mushroom mycelium obtained according to the method of the present invention contains more dietary fiber than the mushroom mycelium obtained by ordinary culture methods that do not contain GABA, etc., so it can also be used as a mushroom containing a large amount of dietary fiber. Food (food fiber fortified food).

[Example]

The present invention will be specifically described below with reference to examples, but the present invention is not limited thereto.

[Example 1]

(Cultivation with GABA added)

<Pre-culture of Basidiomycetes>

Cultivate Pleurotus eryngii on PDA flat culture medium until it reaches full growth.

<Special culture>

Use a sterilized spatula and the medium to scrape a small piece of medium with mycelium grown on the pre-cultured flat medium, and inoculate it into sterilized 100mL liquid medium (glucose 3%, polyprotein 0.5%, yeast extract 0.3%, diphosphate Potassium hydrogen 0.1%, magnesium sulfate 0.05%, Tween80 0.1%), in a flask with a volume of 300 mL, culture it at 28°C and 125 rpm for 7 days to obtain a seed culture medium.

<Main training>

The obtained seed culture liquid was subjected to solid-liquid separation using centrifugal separation (7000 rpm, 10 minutes), and the mycelium was recovered. Physiological saline was added to the recovered mycelium and pulverized using a glass bead mill type homogenizer. Transplant 3 mL of the crushed liquid into 300 mL of main culture medium (glucose 3%, polyprotein 0.5%, yeast extract 0.3%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.05%, Tween80 0.1%, GABA 1% by weight), Cultivate in a 2L Erlenmeyer flask at 28°C and 125 rpm for 7 days.

The liquid at the end of the culture is centrifugally separated into culture liquid and mycelium.

After drying the cultured mycelium, the GABA content in the mycelium was measured and the result was approximately 2000 mg/100g (amino acid analysis using high-speed liquid chromatography).

[Example 2]

(Cultivation with glutamate added)

Seed culture and main culture were performed under the same conditions as in Example 1. However, during main culture, 2 wt% of MSG (sodium L-glutamate) and 0.01 wt% of pyridoxal phosphate were added instead of GABA.

[Example 3]

Seed culture and main culture were performed under the same conditions as in Example 1. However, 2% by weight of MSG (sodium L-glutamate) was added instead of GABA during the main culture.

[Example 4]

(Cultivation with addition of polyamine)

Seed culture and main culture were performed under the same conditions as in Example 1. However, during the main culture, 0.02% by weight of a mixture of putrescine, spermidine and spermine was added instead of GABA.

(Comparative example 1)

No GABA was added during the main culture of Example 1, except that the culture was performed under the same conditions to obtain Pleurotus eryngii mycelium.

The Pleurotus eryngii mycelium obtained during the cultivation of Examples 1 to 4 and Comparative Example 1 was dried by freeze-drying. As a result, in Examples 1 and 2, 28 g/L of mycelium was obtained in terms of dry bacterial cells. Examples 3 and 4 can obtain 24.4g/L mycelium based on dry bacterial cells. On the other hand, Comparative Example 1 was 11 g/L.

The mushroom mycelium obtained in Examples 1 and 2 produced red pigment, and both the mycelium and the culture solution had a red appearance. By the method of Comparative Example 1, white mycelium like the fruiting body of Pleurotus eryngii was obtained.

The dietary fiber in the mycelium obtained in Example 1, Example 2, and Comparative Example 1 was measured. For reference, the dietary fiber content of Pleurotus eryngii fruiting bodies was also determined. The measurement results are shown in Table 1.

In addition, the measurement method of dietary fiber is based on the enzyme-gravimetric method (using enzymes: α-amylase, aminoglucosidase (Aminoglucosidase), and protease).

Pleurotus lunge (NBRC30791 strain) was used instead of Pleurotus eryngii used in Examples 1, 2 and Comparative Example 1, and the effect of adding GABA and the like was confirmed. The yield of each mycelium dried by freeze-drying in the same manner as Pleurotus eryngii mycelium was 21g/L when GABA was added, and 22g/L when glutamic acid was added. In the case of comparison medium, it is 15g/L.

In this way, if culture medium containing γ-aminobutyric acid (GABA), GABA precursors, or substances that produce GABA through metabolism is used for culture, hyphae can be obtained with better efficiency than previous techniques. body. In addition, since the Pleurotus eryngii mycelium and the like of the present invention contain a large amount of GABA, they can also be used as functional foods having the functionality of GABA.

Claims (3)

A method for manufacturing mushroom mycelium, including the following steps: cultivating in a culture medium containing γ-aminobutyric acid (GABA) to form mushroom mycelium; the content of the γ-aminobutyric acid (GABA) is In terms of solid content, it is 0.01 to 10% by weight. The mushroom mycelium system belongs to the Basidiomycetes of the phylum Basidiomycota. A composition containing: mushroom mycelium cultured in a medium containing γ-aminobutyric acid (GABA); the content of the γ-aminobutyric acid (GABA), in terms of solid content, is 0.01~10% by weight, the mushroom mycelium system belongs to Basidiomycetes of the phylum Basidiomycota. A food containing the composition of claim 2.