JP6087115B2 - Ornithine-containing food material using dried wheat shochu for basidiomycete culture - Google Patents

Description

  The present invention relates to a food composition containing a high amount of ornithine and a method for producing the same.

  It is known that fruit bodies (mushrooms) formed by basidiomycetes produce a large amount of useful secondary metabolites such as β-glucan and protease (Non-patent Documents 1 and 2). It has also been reported that β-glucan and lactic acid bacteria have an immunostimulatory effect, stimulate cells such as NK cells and killer T cells, and attack cancer cells.

  It has been reported that amino acids including ornithine and γ-aminobutyric acid increase by freezing fruit bodies such as enokitake mushrooms, beech shimeji mushrooms and eringgi in fungal bed cultivation (Patent Documents 1 and 2). It has been reported that ornithine has an effect of enhancing liver function and motility, and γ-aminobutyric acid has an effect of improving hypertension.

  However, the amount of free amino acids (ornithine etc.) in fruit bodies obtained in mushroom bed cultivation is too small to produce free amino acids at low cost and in large quantities. A method for producing ornithine by liquid culture using bacteria having ornithine-producing ability is known, but since a medium containing components that are not suitable for food is used, it cannot be used as a food composition as it is, and is safe. In order to ensure the above, it is necessary to purify ornithine from the culture (Patent Documents 1, 2, 3, and Non-Patent Document 3). In that case, since the cost is required for the purification process, the production cost of ornithine increases.

JP 2007-53924 A JP 2007-274904 A JP 2005-102559 A

Yutaka Yamada, Emiko Nakayama et al., Enzyme profiles of basidiomycetes and their bagasse degradation process, Grassland Science, 2000, 46, p.265 Susumu Hirase, Sachiko Nakai, Chemical structure of antitumor polysaccharide of basidiomycete Kawaratake, YAKUGAKUZASSHI, 1976, 96, p.419 Masaki Sakakibara, High content of γ-aminobutyric acid (GABA) and blood pressure lowering effect by lactic acid fermentation of spiruna, DIC Technical Review, 2006, No. 12, p.13

  An object of the present invention is to provide an efficient method for producing a food composition containing a high amount of ornithine.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that fruit bodies obtained by fungal bed cultivation using a medium containing dry barley shochu or a fungus bed immediately after the formation of the fruit body primordia It has been found that a large amount of free arginine accumulates, and that a food composition containing a high amount of ornithine converted from free arginine can be obtained by seeding and culturing lactic acid bacteria on the fruit body or fungus bed. The invention has been completed.

That is, the present invention includes the following.
[1] Ornithine comprising culturing edible basidiomycetes in a fungus bed medium containing dry wheat shochu, collecting fruit bodies or fungus beds containing the produced free arginine, and seeding and cultivating lactic acid bacteria A method for producing a food composition comprising
In this method, the bacterial bed medium preferably contains a food containing a high content of arginine. Suitable examples of foods high in arginine are foods derived from beans, seeds, or cereals.
In this method, it is preferable that the bacterial bed on which the lactic acid bacteria are seeded is a bacterial bed up to 3 days after the formation of the fruit body primordium.
The edible basidiomycete used in this method is more preferably an edible basidiomycete belonging to the order Agaric, for example, the genus Oleander or Enokitake. For example, the edible basidiomycete may be oyster mushrooms or enokitake mushrooms.
The lactic acid bacterium used in this method is more preferably Lactobacillus or Pediococcus. This lactic acid bacterium is particularly preferably Lactobacillus fermentum or Pediococcus pentosaceus. In a preferred embodiment, the lactic acid bacterium is a Lactobacillus fermentum Kirishima 1R (NITE P-784) strain or a Pediococcus pentosaceus Kirishima 1C (NITE P-787) strain.
The food composition containing ornithine obtained by this method preferably also contains lactic acid bacteria.
[2] A food composition containing ornithine obtained by the method according to [1] above.
In a preferred embodiment, the food composition contains 500 mg or more of ornithine per 100 g in a dry weight ratio.

  If the method of this invention is used, the food composition which contains ornithine highly can be manufactured efficiently.

  Hereinafter, the present invention will be described in detail.

  In the method according to the present invention, an edible basidiomycete is cultured in a bacterial bed medium containing dry barley shochu. In the present invention, “dry wheat shochu” means that a solid fraction (for example, obtained by solid-liquid separation) separated from shochu distilled spirit produced in the process of producing shochu using wheat as a raw material is dried. A substance. The moisture content of the dried wheat shochu is not particularly limited, but is typically 15% or less, for example, 5 to 10%. The dry wheat shochu is not limited, but is, for example, 5% by weight (wt%) or more, preferably 10% by weight or more, more preferably 20% by weight or more, and further preferably 30% by weight or more in terms of dry weight. Particularly preferably, it is preferably contained in the bacterial bed medium in an amount of 50% by weight or more. The protein produced in the basidiomycete mycelium degrades the protein in the medium, and free arginine is produced from the bound arginine contained in the protein and released into the medium. In the method of the present invention, the production of free arginine by basidiomycetes can be remarkably promoted by using a bacterial bed medium containing dry barley shochu. Here, “linked arginine” is an arginine residue contained in a polypeptide chain composed of two or more amino acid residues, and “free arginine” is such a polypeptide chain. It is an arginine that is present alone without being included in the above.

  In one embodiment of the present invention, it is also preferable that the fungus bed medium containing dried barley shochu further contains a food containing a high arginine content. The high arginine-containing food means a food having a high arginine content in the protein composition of the food, for example, 300 mg / 100 g or more, more preferably 500 mg / 100 g or more, more preferably 1000 mg / 100 g or more, especially in dry weight ratio. A food containing 2000 mg / 100 g or more of arginine (mainly bound arginine) is preferred. For example, beans, seeds, some vegetables, and cereals, as well as meat and gelatin are known to be rich in arginine in their protein composition. As the arginine-rich food used in the present invention, beans, seeds, and cereal-derived foods are particularly preferable. The legume-derived food includes seeds (beans) such as soybean, broad bean, mung bean, azuki bean, cowpea, and pea, or processed foods thereof. Examples of the processed food include, but are not limited to, soybean powder, soybean powder, soybean powder such as soybean powder, defatted soybean, Takano tofu, miso, okara (dried okara, etc.), yuba, etc. It is done. Nuts and seeds-derived foods include seeds of nuts such as peanuts, sesame seeds, almonds, cashew nuts, macadamia nuts and hazelnuts, and seeds such as pine nuts, ginkgo seeds, sunflower seeds and pumpkin seeds, or processed foods thereof. Examples of the processed food include, but are not limited to, dried products obtained by removing seed coats and shells, and dried products thereof. Cereal-derived foods include wheat such as wheat, barley, rye and oats, cereals such as corn, rice, buckwheat, and amaranth or processed foods thereof. Examples of the processed food include, but are not limited to, pulverized cereals such as wheat flour and buckwheat flour, wheat germ, rice bran, bran, brown rice, polished rice, and bread crumbs. Another preferred arginine-rich food includes gelatin. Further preferred arginine-rich foods include garlic, dried shiitake mushrooms, kanpyo and processed foods thereof. The fungus bed medium may contain, for example, 5 to 40% by weight of one or more arginine-rich foods in a total weight ratio. By using a bacterial bed medium containing a food containing a high content of arginine, free arginine can be produced at a higher concentration via the protease activity of basidiomycetes. The present invention also provides such a bacterial bed medium.

  In order to promote the growth of basidiomycetes, the above-mentioned fungus bed medium contains fibrous plant materials such as corn cob, potato starch lees, shell fossil fine powder, oyster shell powder, baking soda, malt extract, yeast extract, glucose, etc. You may make it contain. However, the component added to the above-mentioned fungus bed medium is usually edible. The pH of the bacterial bed medium may be in a range where the basidiomycete used can grow, but is usually pH 5-7.

  The “edible basidiomycete” used in the present invention means a basidiomycete that forms a fruiting body and that the fruiting body is edible. The “edible basidiomycete” may be a white jellyfish, a jellyfish, a hymenoptera, an agaric or the like, and preferably an agaric. The “edible basidiomycetes” may be, for example, oyster mushrooms, agaricaceae, shimeji mushrooms, xylem mushrooms, sarcoma moss mushrooms, agaricaceae mushrooms, oyster mushrooms, mushrooms, shimeji mushrooms, enokitake mushrooms, shiitake mushrooms, maitake mushrooms It may be of the genus Hanabiratake. Specific examples of “edible basidiomycetes” include, but are not limited to, Enokitake (Enokitake), Bunashimeji (Shimeji), Eringi (Pleurotus), Himatsutake (Agaricus), Maitake (Maitake) ), Oyster mushrooms (genus Oyster mushrooms), chanterelles (genus Hanatake mushrooms), etc., which are highly safe and widely edible since ancient times.

  The above-mentioned fungus bed medium is mixed by adding water in an amount of water to the medium material so that the water content of the fungus bed medium is generally 50 to 70% by weight, typically about 65% by weight. Preferably, after mixing uniformly, the container (a bottle, a bag, etc.) is filled and sterilized. The sterilization treatment can be performed by, for example, heating or warming autoclaving, and can be performed by, for example, processing at a temperature exceeding 100 ° C. (eg 121 ° C.) for several hours (eg 3 hours). The fungus bed medium thus prepared is solid and can be inoculated with basidiomycetes after cooling.

  Preferably, the above-mentioned edible basidiomycete is inoculated into the above-mentioned microbial bed medium cooled to 20 ° C. or lower and cultured. The basidiomycete may be cultured under culture conditions (temperature, humidity, light conditions, etc.) suitable for each basidiomycete. The number of culture days varies depending on the temperature, medium composition, basidiomycete type condition, etc., but at least it is necessary that the mycelium grow and the protease is sufficiently produced. In general, the culture period until the fruiting body primordium formation time or the fruiting body formation completion time is about 20 to 90 days from the start of the culture.

  Regarding the culture conditions of various basidiomycetes, for example, “Mushroom Cultivation” (Omori Kiyotosu, edited by Hiroshi Koide, published on September 30, 2001, Rural Fishing Village Cultural Association), “Mushroom Handbook” (Kenjiro Kinugawa) , Shin Ogawa, (2000) Asakura Shoten) and other textbooks on mushroom cultivation. After culture of basidiomycetes over a certain period, bacteria scraping, water injection, etc. may be performed as a fruit body generation treatment.

  As an example, when oyster mushroom is used as an edible basidiomycete, it is cultured for 30 to 35 days under dark conditions as much as possible at a culture temperature of 20 to 23 ° C. and a relative humidity of 65 to 75%. Fruit bodies can be obtained by culturing at 10 ° C. (appropriate temperature: 15 ° C.) and relative humidity of 90 to 95% for 10 to 18 days.

  As another example, when enokitake is used as an edible basidiomycete, it is cultured for 10 to 20 days (for example, 15 days) under dark conditions as much as possible at a culture temperature of 17 to 19 ° C. (preferably 18 ° C.) and a relative humidity of 75 to 85%. After that, the cells are cultured for 18 to 22 days under dark conditions as much as possible at a culture temperature of 19 to 21 ° C. (preferably 20 ° C.) and a relative humidity of 65 to 75%, and then the culture temperature is 15 to 17 ° C. (preferably 16 ° C.). After forming the fruiting body primordium, the fruiting body can be obtained by culturing for 10 to 18 days at a temperature of 10 to 20 ° C. (appropriate temperature: 15 ° C.) and a relative humidity of 90 to 95%.

  By culturing the edible basidiomycetes in the above-mentioned fungus bed medium as described above, first, a fruiting body primordium is formed, and the primordial group grows into a fruiting body (forming a fruiting body). Fruit bodies (mushrooms) can be obtained by such fungus bed cultivation. The fruit bodies thus obtained contain a large amount of free arginine produced by degrading proteins in the medium (for example, those derived from dried barley shochu and / or arginine-rich foods) by the protease produced by basidiomycete mycelium. It is taken in and accumulated. In one embodiment of the method of the present invention, the harvested (collected) fruiting body can be subjected to lactic acid bacteria culture to convert free arginine to ornithine. As a fruit body to be subjected to lactic acid bacteria culture, it is preferable to use a fruit harvested after the fungus has been opened to some extent (usually 60% or more, preferably 80% or more).

  The bacterial bed before and after the formation of the fruiting body primordium also contains a large amount of the produced free arginine. The concentration of free arginine in the fungal bed increases over time until the formation of the fruiting body primordium, but as the fruiting body primordium grows into the fruiting body, the free arginine moves to the fruiting body, Free arginine concentration decreases. For this reason, the mycelium immediately after the formation of the fruiting body primordium in a stage where the mycelium is sufficiently spread without generating fruiting bodies contains the highest concentration of free arginine. Therefore, in another embodiment of the method of the present invention, the bacterial bed before and after the formation of fruiting body primordium can be collected and subjected to lactic acid bacteria culture to convert free arginine to ornithine. The microbial bed used for lactic acid bacteria culture is preferably a microbial bed 7 to 7 days after the formation of fruiting body primordium, more preferably a bed from 3 to 3 days after the formation of fruiting body primordium. More preferably, the bacterial bed is from 1 to 1 day after the formation of the fruiting body primordium observed by

  As a preferred example, when oyster mushroom is used as an edible basidiomycete, it is cultured at a culture temperature of 20 to 23 ° C. and a relative humidity of 65 to 75 ° C. under dark conditions as much as possible for 30 to 35 days. (A suitable temperature: 15 ° C.) By culturing at a relative humidity of 90 to 95% until just before the formation of the fruiting body primordium, a bacterial bed to be subjected to lactic acid bacteria culture can be prepared.

  As another suitable example, when enokitake is used as an edible basidiomycete, the culture temperature is 17 to 19 ° C. (preferably 18 ° C.), the relative humidity is 75 to 85%, and the dark condition is as much as possible for 10 to 20 days (for example, 15 Culturing for 18 to 22 days (for example, 20 days) under dark conditions as much as possible at a culture temperature of 19 to 24 ° C. (preferably 23 ° C.) and a relative humidity of 65 to 75%. By lowering to 17 ° C. (preferably 15 to 16 ° C.) and culturing until just before the formation of fruiting body primordium, a bacterial bed to be subjected to lactic acid bacteria culture can be prepared.

  The fruiting body or fungus bed containing free arginine at a high concentration obtained as described above preferably has a pH of 6 to 7 when subjected to lactic acid bacteria culture. The pH of the fruiting body is usually about 6 and can be used for lactic acid bacteria culture without adjusting the pH, but the pH may be adjusted. Since the pH of the fungus bed after basidiomycete culture is usually lowered, it is preferable to adjust the pH to 6-7. The pH can be adjusted by a conventional method, but it is preferable to add an appropriate amount of baking soda.

  When using fruit bodies, it is preferable to add lactic acid bacteria to the crushed material obtained by pasting or grinding the fruit bodies with a mixer or the like, mixing (preferably uniformly stirring and mixing), and culturing. . When using a fungus bed, if necessary, crush the fungus bed, add an appropriate amount (for example, 0.5%) of baking soda to the fungus bed, adjust the pH, add lactic acid bacteria, and mix (preferably Are uniformly stirred and mixed) and solid culture is preferable.

  The lactic acid bacterium to be seeded may be any lactic acid bacterium having the ability to convert free arginine to ornithine. Many types of such lactic acid bacteria have been reported, and their conversion ability is widely recognized in lactic acid bacteria. Such lactic acid bacteria to be seeded include, for example, the genus Lactobacillus, Lacococcus, Pediococcus, Lueconostoc, Streptococcus and Enterococcus But is not limited to these genera and species. Examples of the genus Lactobacillus include Lactobacillus fermentum, Lactobacillus plantam, Lactobacillus brevis, Sractobacillus delbruecki, Lactobacillus casei, Lactobacillus pentosas and the like. Examples of the genus Lactococcus include Lactococcus and Lactis cremoris. Examples of the genus Pediococcus include Pediococcus pentosaceus, Pediococcus acidilactici, Pediococcus heparinus and the like. Examples of the genus Leuconostoc include Leuconostoc lactis and Leuconostoc mesenteroides. Streptococcus genus includes Streptococcus thermophilus. Examples of the genus Enterococcus include Enterococcus faecalis and Enterococcus faecium. The lactic acid bacteria used in the method of the present invention are preferably edible. Lactobacillus or Pediococcus is more preferable as the lactic acid bacterium used in the method of the present invention, and for example, Lactobacillus fermentum, Pediococcus pentosaceus and Pediococcus acidilacticis are preferable.

  Examples of lactic acid bacteria that can be preferably used include, but are not limited to, Lactobacillus fermentum Kirishima 1R (NITE P-784) strain, Pediococcus pentosaceus Kirishima 1C (NITE P-787) strain, Lactobacillus -Fermentum Kirishima 3R (NITE P-786) strain, Pediococcus acidilactici Kirishima 2C (NITE P-788) strain, and the like. These exemplified lactic acid strains (NITE P-784, NITE P-787, NITE P-786, and NITE P-788) are registered with the Patent Microorganism Depositary Center of Product Evaluation Technology (Kazusa, Kisarazu, Chiba, Japan). It has been deposited on July 23, 2009 at Kamaashi 2-5-8).

Lactic acid bacteria are not limited to fruiting bodies or cells, but are added in an amount of 1.0 × 10 ⁵ cfu / g or more (total weight ratio), for example, 1.0 to 9.0 × 10 ⁶ cfu / g. It is preferable to do.

The lactic acid bacteria culture may be performed under culture conditions suitable for the lactic acid bacteria used. As a preferred example, after lactic acid bacteria are uniformly stirred and mixed with fruit bodies or cells, they may be cultured at 30 to 40 ° C. (for example, 35 ° C.) for 15 to 96 hours (for example, 18 to 30 hours). In general, when free arginine contained in fruiting bodies or cells is converted to ornithine by the method of the present invention, lactic acid bacteria culture can be completed in about 20 hours. In a typical example, the pH after culturing is about 5.0, and the bacterial cell concentration of lactic acid bacteria is 1.0 × 10 ⁹ cfu / g or more.

  A food composition (food material) containing ornithine can be prepared from the culture after culturing lactic acid bacteria thus obtained. The culture can be used as it is as a food composition, or it can be processed into a food composition in a desired form. The present invention also provides such a food composition. For example, the culture after culturing lactic acid bacteria may be dried. The drying step can be performed using any drying technique such as hot air drying, air drying, low temperature drying, vacuum drying, spray drying, freeze drying, and the like. In a specific example, for example, the culture product after culturing lactic acid bacteria may be blown and dried at 40 to 50 ° C. The culture after culturing lactic acid bacteria or a dried product thereof may be processed by pulverization (for example, mill pulverization), concentration, pasting, dilution, tableting, or encapsulation. If necessary, it is also preferable to perform food processing after removing a relatively hard material (such as corn cob) contained in the bacterial bed medium from the culture after culturing lactic acid bacteria (such as by a sieve). The food composition according to the present invention is not limited to the following, but may be in any form such as powder, granule, solid material, solution, suspension, gel, tablet, capsule and the like. The food composition according to the present invention contains ornithine in a high concentration, for example, 500 mg or more, preferably 900 mg or more, more preferably 1000 mg or more, more preferably 1500 mg or more, particularly preferably 1500 mg or more, per 100 g of the food composition in a dry weight ratio. Contains over 2000 mg of ornithine. Since ornithine has known effects such as protection and improvement of liver function and improvement of motility, the food composition according to the present invention can also be used as a functional food.

  In addition to ornithine, the food composition according to the present invention preferably contains lactic acid bacteria (live cells and dead cells) used for lactic acid bacteria culture and crushed materials thereof. Lactic acid bacteria killed cells or crushed materials thereof are also known to have immunostimulatory effects, and food compositions containing them can be expected to have functionality such as immunostimulation. The amount of lactic acid bacteria contained in the food composition varies depending on the applied food processing, but is basically proportional to the number of bacteria after culture. The food composition according to the present invention may contain, in addition to ornithine and lactic acid bacteria, other free amino acids produced together with arginine during bacterial bed culture and other components such as produced β-glucan at higher concentrations. . The food composition according to the present invention also contains a free arginine produced during the fungal bed culture that has not been converted to ornithine.

  The food composition according to the present invention may further contain various additives that are conventionally used in food and drink. Additives include, but are not limited to, colorants (gardeny pigments, red 102, etc.), fragrances (orange fragrances, etc.), sweeteners (sucrose, stevia, aster palm, etc.), preservatives (sorbic acid Etc.), emulsifiers (such as propylene glycol fatty acid esters), antioxidants (such as vitamin C), pH adjusters (such as citric acid), thickeners (such as xanthan gum), swelling agents (such as calcium carbonate), defoamers (such as Calcium phosphate), binders (such as sodium polyphosphate), nutrient enhancers (calcium enhancer, vitamin A, etc.), excipients (water-soluble dextrin, etc.) and the like.

  The food composition according to the present invention may be used for human food, but includes livestock (cattle, horses, sheep, goats, pigs, camels, chickens, ducks, etc.) and pet / laboratory animals (rats, mice, It may be used for the feed of non-human animals such as rodents such as gerbils, hamsters, and guinea pigs, ferrets, rabbits, dogs, cats, minipigs, monkeys, and the like.

  Hereinafter, the present invention will be described more specifically with reference to examples. However, the technical scope of the present invention is not limited to these examples.

(Example 1) Analysis of arginine production and accumulation amount in oyster mushroom and enokitake mushroom beds According to the formulation of Table 1, a fungal bed medium (dried wheat shochu mixed medium) was prepared. The pH of the medium was adjusted to around 5.5 with shell fossils (fine powder). The raw materials shown in Table 1 were mixed according to the stated ratio, water was added so that the water content was 65% (total weight ratio), and the mixture was stirred and mixed. This was put into a sterilization kettle and sterilized by heating at 121 ° C. for 3 hours. After the temperature of the fungus bed medium was cooled to 20 ° C. or lower, oyster mushroom seeds were inoculated into the medium. This was cultured for 30-35 days under dark conditions at a temperature of 20-23 ° C., a relative humidity of 65-75%. Subsequently, further cultivation was performed at a relative humidity of 90 to 95% and a temperature of 10 to 20 ° C. (appropriate temperature of 15 ° C.) for about 2 weeks to obtain fruit bodies.

  Similarly, enokitake was cultured using a fungus bed medium having the formulation shown in Table 1. The raw materials shown in Table 1 were mixed according to the stated ratio, water was added so that the water content was 65% (total weight ratio), and the mixture was stirred and mixed. This was put into a sterilization kettle and sterilized by heating at 121 ° C. for 3 hours. After the temperature of the fungus bed medium was cooled to 20 ° C. or lower, enokitake mushroom seeds were inoculated into the medium. The first 15 days (initial) were at a temperature of 18 ° C. and a relative humidity of 80%, and the following 20 days (late stage) were at a temperature of 20 ° C. and a relative humidity of 70%. Thereafter, the temperature was maintained at 16 ° C. until just before the formation of the fruiting body primordium. After the fruit body primordium formation, the fruit body was further cultured for about 2 weeks at a relative humidity of 90 to 95% and a temperature of 10 to 20 ° C. (appropriate temperature of 15 ° C.).

  During cultivation, the mycelia and enokitake mushroom beds were collected over time until the fruit bodies were formed after the mycelium was spread (spread) in the medium, and the arginine concentration in the fungus bed was measured. First, the collected microbial bed was pulverized, a fixed amount of pulverized material was collected, adjusted to pH 2.2, and then finely pulverized with a homogenizer to extract amino acids. Subsequently, the extract was centrifuged (at 4,000 rpm for 8 minutes), the supernatant was collected, and after membrane filtration, amino acid analysis (Shimadzu high performance liquid chromatograph LC-VP) was performed.

  A summary of the results of arginine concentration measurement is shown in Table 2. As shown in Table 2, oyster mushrooms and enokitake mushrooms both produced and secreted a large amount of arginine in the fungus bed in the dry wheat shochu mixed fungus bed medium. It was also shown that the free arginine concentration in the fungus bed was maximum immediately after the formation of the fruiting body primordium.

  Proteases in the basidiomycete hyphae degrade proteins and produce free amino acids. As shown in Table 2, the free arginine concentration in the fungus bed reached a maximum immediately after the formation of the fruiting body primordium, so that the free arginine produced from the protein by the protease in the mycelium is It was shown that it migrates to the entity and accumulates. In addition, when arginine in the bacterial bed is converted to ornithine by lactic acid bacteria culture, it has been shown that it is particularly advantageous to use the bacterial bed immediately after the formation of the fruiting body primordium in order to maximize the ornithine concentration.

(Example 2) Measurement of free arginine concentration in oyster mushroom fruit body and ornithine concentration after cultivation of lactic acid bacteria using fruit body A bacterial bed medium was prepared according to the formulation of each medium shown in Table 3. The raw materials of each medium were mixed according to the stated ratio, water was added so that the water content was 65% (total weight ratio), and the mixture was stirred and mixed. This was put into a sterilization kettle and sterilized by heating at 121 ° C. for 3 hours. After the temperature of the fungus bed medium was cooled to 20 ° C. or lower, oyster mushroom seeds were inoculated into the medium. This was cultured for 30-35 days under dark conditions at a temperature of 20-23 ° C., a relative humidity of 65-75%. Subsequently, further cultivation was performed at a relative humidity of 90 to 95% and a temperature of 10 to 20 ° C. (appropriate temperature of 15 ° C.) for about 2 weeks to obtain fruit bodies.

  Subsequently, the arginine concentration in the fruit body was measured. First, the collected fruit body was pulverized, a certain amount of pulverized material was collected, adjusted to pH 2.2, and then finely pulverized with a homogenizer to extract amino acids. Subsequently, the extract was centrifuged (at 4,000 rpm for 8 minutes), the supernatant was collected, and after membrane filtration, amino acid analysis (Shimadzu high performance liquid chromatograph LC-VP) was performed. The results of measurement of arginine concentration are shown in Table 4.

Furthermore, the obtained oyster mushroom fruit body was crushed with a mixer to form a paste. Lactic acid bacterium Lactobacillus fermentum Kirishima 1R was added (seeded) to 10 ⁶ cfu / g or more. After stirring and mixing uniformly, the mixture was cultured at 35 ° C. for 20 hours. After culturing, the culture was blown and dried at 45 ° C. and pulverized. A certain amount of pulverized material was collected, adjusted to pH 2.2, and then finely pulverized with a homogenizer to extract amino acids. Next, the extract was centrifuged (4,000 rpm for 8 minutes), the supernatant was collected, and after membrane filtration, amino acid analysis (Shimadzu high performance liquid chromatograph LC-VP) was performed to measure the ornithine concentration (Table 4).

  In addition, lactic acid bacteria were extracted from the culture after culturing lactic acid bacteria with physiological saline, and the bacterial cell concentration was measured by an agar dilution plate method using MRS medium (Table 4).

  As shown in Table 4, the arginine concentration in the fruit body cultured in the test medium H1 (containing 50% by weight of dry wheat shochu) was 2,934 mg / 100 g, and the ornithine concentration after lactic acid bacteria culture was 2,760 mg / 100 g. And increased 5 to 6 times or more compared to the case of the standard medium H5 (50% by weight of rice bran). The arginine concentrations in fruit bodies cultured in the test medium H2 (containing 25% by weight of dry wheat shochu) and test medium H3 (containing 10% by weight of dry barley shochu) are 2,354 mg / 100 g and 2,067 mg / kg, respectively. The ornithine concentration after cultivation of lactic acid bacteria was 2,188 mg / 100 g and 1,888 mg / 100 g, respectively, and increased by a factor of 4 or more compared to the standard medium H5 (containing 50% by weight of rice bran). . On the other hand, the arginine concentration in fruit bodies cultured in the comparative medium H4 (containing 50% by weight of dried sweet potato shochu) is 1,220 mg / 100 g, and the ornithine concentration after lactic acid bacteria culture is 1,043 mg / 100 g. Compared to the case of the standard medium H5 (containing 50% by weight of rice bran), the increase was about twice as much. A dried pulverized product of a culture product of oyster mushroom fruit bodies cultured in a wheat shochu mixed medium contains a high concentration of ornithine and can be used as a useful amino acid-containing food composition (food material).

All the lactic acid bacteria after culturing lactic acid bacteria using fruiting bodies were on the order of 10 ⁹ cfu / g, and the number of bacteria was very high.

(Example 3) Measurement of free arginine concentration in oyster mushroom bed and ornithine concentration after cultivation of lactic acid bacteria using fungus bed. A bed medium was prepared according to the formulation of each medium shown in Table 3. The raw materials of each medium were mixed according to the stated ratio, water was added so that the water content was 65% (total weight ratio), and the mixture was stirred and mixed. This was put into a sterilization kettle and sterilized by heating at 121 ° C. for 3 hours. After the temperature of the fungus bed medium was cooled to 20 ° C. or lower, oyster mushroom seeds were inoculated into the medium. This was cultured for 30-35 days under dark conditions at a temperature of 20-23 ° C., a relative humidity of 65-75%. Subsequently, the cells were cultured at a relative humidity of 90 to 95% and at a temperature of 10 to 20 ° C. (appropriate temperature of 15 ° C.) until just before the fruiting body primordium formation, and the culture was completed immediately after the primordium formation.

  Subsequently, the concentration of arginine in the fungus bed after completion of the culture (oyster mushroom fungus bed) was measured. First, the collected microbial bed was pulverized, a fixed amount of pulverized material was collected, adjusted to pH 2.2, and then finely pulverized with a homogenizer to extract amino acids. Subsequently, the extract was centrifuged (at 4,000 rpm for 8 minutes), the supernatant was collected, and after membrane filtration, amino acid analysis (Shimadzu high performance liquid chromatograph LC-VP) was performed. The results of arginine concentration measurement are shown in Table 5.

Furthermore, 0.5% of sodium bicarbonate was added to the collected oyster mushroom beds to adjust the pH to about 6.5, and then lactic acid bacteria Lactobacillus fermentum Kirishima 1R was added so as to be at least 10 ⁶ cfu / g (seeding). )did. After stirring and mixing uniformly, the mixture was cultured at 35 ° C. for 20 hours. After culturing, the culture was blown and dried at 45 ° C. and pulverized. A certain amount of pulverized material was collected, adjusted to pH 2.2, and then finely pulverized with a homogenizer to extract amino acids. Next, the extract was centrifuged (4,000 rpm for 8 minutes), the supernatant was collected, and after membrane filtration, amino acid analysis (Shimadzu high performance liquid chromatograph LC-VP) was performed to measure the ornithine concentration (Table 5).

  In addition, lactic acid bacteria were extracted from the culture after culturing lactic acid bacteria with physiological saline, and the bacterial cell concentration was measured by an agar dilution plate method using MRS medium (Table 5).

  As shown in Table 5, the arginine concentration in the fungus bed cultured in the test medium H1 (containing 50% by weight of dried wheat shochu) and the ornithine concentration after culturing lactic acid bacteria are in the case of the standard medium H5 (containing 50% by weight of rice bran) It increased 6 to 7 times or more. The arginine concentration in the fungus bed cultured in the test medium H2 (containing 25% by weight of dried wheat shochu) and the ornithine concentration after culturing lactic acid bacteria were also 5-6 compared to the case of the standard medium H5 (containing 50% by weight of rice bran). More than doubled. The arginine concentration in the fungus bed cultured in the test medium H3 (containing 10% by weight of dried wheat shochu) and the ornithine concentration after culturing lactic acid bacteria were also 4-5 compared with the standard medium H5 (containing 50% by weight of rice bran). More than doubled. On the other hand, the arginine concentration in the fungus bed cultured in the comparison medium H4 (containing 50% by weight of dried sweet potato shochu) and the ornithine concentration after culturing lactic acid bacteria were 1,043 mg / 100 g, and the standard medium H5 (containing 50% by weight of rice bran) ) Increased only about twice as much as in the case of). The dried pulverized product of the cultured oyster mushroom bed after cultivating oyster mushrooms in a barley shochu mixed medium contains a high concentration of ornithine and can be used as a useful amino acid-containing food composition (food material). it can.

All of the lactic acid bacteria after culturing lactic acid bacteria using the fungus bed were on the order of 10 ⁹ cfu / g, and the number of bacteria was very high.

(Example 4) Measurement of free arginine concentration in enokitake mushroom fruit body and ornithine concentration after lactic acid bacteria culture using fruit body A bacterial bed medium was prepared according to the formulation of each medium shown in Table 6. The raw materials of each medium were mixed according to the stated ratio, water was added so that the water content was 65% (total weight ratio), and the mixture was stirred and mixed. This was put into a sterilization kettle and sterilized by heating at 121 ° C. for 3 hours. After the temperature of the fungus bed medium was cooled to 20 ° C. or lower, enokitake mushroom seeds were inoculated into the medium. The first 15 days (initial) were at a temperature of 18 ° C. and a relative humidity of 80%, and the following 20 days (late stage) were at a temperature of 20 ° C. and a relative humidity of 70%. Thereafter, the temperature was maintained at 16 ° C. until just before the formation of the fruiting body primordium. After the fruit body primordium formation, the fruit body was obtained by further culturing at a relative humidity of 90 to 95% and a temperature of 10 to 20 ° C. (appropriate temperature of 15 ° C.) for about 2 weeks.

  Subsequently, the arginine concentration in the fruit body was measured in the same manner as in Example 2. The results of arginine concentration measurement are shown in Table 7.

Furthermore, in the same procedure as in Example 2, the lactic acid bacteria Pediococcus pentosaceus Kirishima 1C was added (seeded) so as to be 10 ⁶ cfu / g or more to the crushed enokitake fruit body, and lactic acid bacteria culture was performed. It was. After culturing, the culture was blown and dried at 50 ° C. and pulverized. The pulverized product was subjected to amino acid analysis (Shimadzu high performance liquid chromatograph LC-VP) in the same manner as in Example 2 to measure the ornithine concentration (Table 7).

  Moreover, about the culture after lactic acid bacteria culture | cultivation, it carried out similarly to Example 2, and measured the bacterial count density | concentration of lactic acid bacteria (Table 7).

  As shown in Table 7, the arginine concentration in the fruit body cultured in the test medium E1 (containing 30% by weight of dried wheat shochu) and the ornithine concentration after cultivation of lactic acid bacteria are in the case of the standard medium E4 (containing 30% by weight of rice bran) Each increased by more than 5 times. The arginine concentration in the fruit body cultured in the test medium E2 (containing 10% by weight of dried wheat shochu) and the ornithine concentration after culturing lactic acid bacteria were 3-4 compared to the case of the standard medium E4 (containing 30% by weight of rice bran). More than doubled. On the other hand, the arginine concentration in the fruit body cultured in the comparative medium E3 (containing 30% by weight of dried sweet potato shochu) and the ornithine concentration after cultivation of lactic acid bacteria were 2 as compared with the case of the standard medium E4 (containing 30% by weight of rice bran). The increase was about double. The dried and pulverized product of the enokitake mushroom fruit body cultured after lactic acid bacteria culture in a wheat shochu mixed medium contains a high concentration of ornithine and can be used as a useful amino acid-containing food composition (food material).

All the lactic acid bacteria after culturing lactic acid bacteria using fruiting bodies were on the order of 10 ⁹ cfu / g, and the number of bacteria was very high.

Example 5 Measurement of Free Arginine Concentration in Enokitake Bacteria Bed and Ornithine Concentration after Lactic Acid Bacteria Culture Using Bacteria Bed A bacterial bed medium was prepared according to the formulation of each medium shown in Table 6. The raw materials of each medium were mixed according to the stated ratio, water was added so that the water content was 65% (total weight ratio), and the mixture was stirred and mixed. This was put into a sterilization kettle and sterilized by heating at 121 ° C. for 3 hours. After the temperature of the fungus bed medium was cooled to 20 ° C. or lower, enokitake mushroom seeds were inoculated into the medium. The first 15 days (initial stage) were at a temperature of 18 ° C. and a relative humidity of 80%, and the following 20 days (late stage) were at a temperature of 23 ° C. and a relative humidity of 70%. Thereafter, the temperature was maintained at 15 ° C. until just before the formation of the fruiting body primordium. The culture was completed immediately after the formation of the primordium.

  Subsequently, the arginine concentration in the fungus bed (enokitake mushroom bed) after completion of the culture was measured in the same manner as in Example 3. The results of arginine concentration measurement are shown in Table 8.

Further, in the same manner as in Example 3, 0.5% sodium bicarbonate was added to the obtained enokitake mushroom bed to adjust the pH to about 6.5, and then the lactic acid bacterium Pediococcus pentosaceus Kirishima 1C was added at 10 ⁶ cfu / It was added (seeded) so as to be g or more, and lactic acid bacteria culture was performed. After culturing, the culture was blown and dried at 45 ° C. and pulverized. About the obtained ground material, it carried out similarly to Example 3, and measured the ornithine density | concentration by the amino acid analysis (Shimadzu high performance liquid chromatograph LC-VP) (Table 8). Moreover, about the culture after lactic acid bacteria culture | cultivation, it carried out similarly to Example 3, and measured the bacterial count density | concentration of lactic acid bacteria (Table 8).

  As shown in Table 8, the arginine concentration in the fungus bed cultured on the test medium E1 (containing 30% by weight of dried wheat shochu) and the ornithine concentration after culturing lactic acid bacteria are in the case of the standard medium E4 (containing 30% by weight of rice bran) It increased 8 to 9 times or more. The arginine concentration in the fungus bed cultured in the test medium E2 (containing 10% by weight of dried wheat shochu) and the ornithine concentration after cultivation of lactic acid bacteria are 6 times or more compared to the standard medium E4 (containing 30% by weight of rice bran). Increased to. On the other hand, the arginine concentration in the fruit body cultured in the comparative medium E3 (containing 30% by weight of dried sweet potato shochu) and the ornithine concentration after cultivation of lactic acid bacteria were 2 as compared with the case of the standard medium E4 (containing 30% by weight of rice bran). The increase was about double. The dried pulverized product of the enokitake mushroom bed after cultivating enokitake mushroom in a shochu-mixed medium contains a high concentration of ornithine and can be used as a useful amino acid-containing food composition (food material). .

All the lactic acid bacteria after culturing lactic acid bacteria using fruiting bodies were on the order of 10 ⁹ cfu / g, and the number of bacteria was very high.

  The method of the present invention is highly safe and can be used for inexpensively producing a food composition (food material) containing free amino acids and the like, particularly ornithine, at a high concentration.

Claims (11)

  Contains ornithine, including cultivating edible basidiomycetes in a fungus bed medium formulated with dry barley shochu, collecting fruit bodies or fungus beds containing the produced free arginine, seeding and cultivating lactic acid bacteria A method for producing a food composition.   The method according to claim 1, wherein the fungal bed medium contains a food containing a high content of arginine.   The method according to claim 2, wherein the arginine-rich food is a food derived from beans, seeds, or grains.   The method according to any one of claims 1 to 3, wherein the microbial bed to be seeded with lactic acid bacteria is a microbial bed from the fruiting body primordium formation to 3 days later.   The method according to any one of claims 1 to 4, wherein the edible basidiomycete is an edible basidiomycete belonging to the order Agaric.   The method according to claim 5, wherein the edible basidiomycete is genus Oyster mushroom or Enokitake mushroom.   The method according to claim 6, wherein the edible basidiomycete is oyster mushroom or enokitake mushroom.   The method according to any one of claims 1 to 6, wherein the lactic acid bacterium is Lactobacillus or Pediococcus.   The method according to claim 8, wherein the lactic acid bacterium is Lactobacillus fermentum or Pediococcus pentosaceus.   The method according to claim 9, wherein the lactic acid bacterium is Lactobacillus fermentum Kirishima 1R (NITE P-784) strain or Pediococcus pentosaceus Kirishima 1C (NITE P-787) strain.   The method according to any one of claims 1 to 10, wherein the food composition containing ornithine contains lactic acid bacteria.