KR20220080627A - Methods for cultivating Basidiomycota using chestnut yulphi and Mushroom Cultivation Method Using Chestnut Yulpi - Google Patents

Abstract

The present invention provides a method for culturing basidiomycetes using chestnut yulpi and a method for cultivating mushrooms using chestnut yulpi, which can increase the taste, quality, and yield of mushrooms by cultivating basidiomycetes using polyphenols.

Description

{Methods for cultivating Basidiomycota using chestnut yulphi and Mushroom Cultivation Method Using Chestnut Yulpi}

The present invention relates to a method for culturing basidiomycetes using chestnut yulpi and a method for cultivating mushrooms using chestnut yulpi.

The reason we can buy Basidiomycetes easily and cheaply at supermarkets is that the cultivation technology of Basidiomycetes has been leveled upward, so the production and quality are similar. This is because the price is also universally downward,

The general method of cultivation of basidiomycetes is to mix sawdust and cotton wool with a nutrient source, sterilize them, cool them, inoculate them, and then temperature-cultivate them. In spring and autumn, when the temperature is low, natural cultivation goes through a natural temperature cultivation process to a facility house, and depending on the natural temperature in spring and autumn, mushrooms are generated, harvested, and shipped.

Among the general cultivation methods of basidiomycetes, the factors affecting the quality of mushrooms are mainly raw materials and nutrients, and accordingly, the technology to increase the production of mushrooms is almost universal and has been accumulated.

Another method is a liquid culture method of basidiomycete culture. The method of culturing basidiomycetes in liquid is mainly mixing various extracted additives in potato medium and inoculating and culturing mushroom bacteria after sterilization. Liquid culture is used to extract specific components, and the cultured components are extracted and used as raw materials or additives for prototypes.

In the existing basidiomycete mushroom cultivation, raw materials such as Douglas pine sawdust stored in stock for more than 6 months, broadleaf sawdust, cottonseed hide or rice straw are mainly used. Nutrient sources include soybean meal, cottonseed meal, soybean husk, corn husk, jade hull, and beet pulp. , jade germ meal, rice bran, beer gourd, liquor gourd and other agricultural by-products are properly mixed and used.

As the raw material for mushroom cultivation, relatively inexpensive conifer sawdust is used, and due to the anti-corrosion component, it has to be stored for more than 6 months, so it takes a lot of space and cost. It is difficult to grow mushrooms consistently because the pH changes each time the medium is mixed, and the production of cotton wool is not constant due to the large differences in pesticide detection and quality.

The nutritional components of the materials used for mushroom cultivation are theoretically compared according to the content of the ingredients influencing the raw materials based on the C/N ratio. Although we are making cultivation media, it is difficult for even skilled technicians to stably produce good quality mushrooms in large quantities because the raw materials for the medium are not uniform every time they are purchased.

For liquid culture of basidiomycetes, most of the large-scale mushroom growers set up facilities for the use of seedlings, and the raw material used is potato extract as the main raw material, and various additives are used.

Commercial basidiomycetes liquid culture users do not go to mushroom cultivation, but use the liquid culture characteristics of rapid mycelial propagation to cultivate basidiomycetes in a liquid state.

Basidiomycete mushroom cultivation is cultivated with high-level technology as described above, but due to overproduction, sales are formed at low unit prices, making it difficult to achieve profits. A differentiated new technology or method is needed to solve this difficult problem.

In order to realize a new technique or method, the patent applicant has used the properties of chestnut yulpi.

When mushrooms are cultivated using chestnut yulpi, the mycelial density and the vitality of bacteria are significantly increased, and the occurrence rate of mushrooms with a low incidence rate is increased.

*Reference

Tannins of polyphenols are water-soluble polyphenolic compounds produced by plants (Barbehenn and Peter Constabel, 2011), and can be subdivided into hydrolyzable (gallotannin and ellagitannins), condensed and complex tannins (Khanbabaee and Ree, 2001).

The tannins of polyphenols still do not have sufficient evidence for the actual functions of tannins and polyphenols in plants, and the information obtained from studies on the reactivity of tannins can be applied to the development of functional substances such as oil-soluble polyphenols.

The method of using condensed tannins has been studied by many scholars, but it is still limited to using hydrolyzable tannins, and there are no papers or patents using condensed tannins directly.

If basidiomycetes are cultivated using condensed tannins, the basidiomycetes decompose the condensed tannins and can be applied to industries in the form of usable water-soluble polyphenols.

Republic of Korea Patent Publication No. 10-2015-0118378

Khanbabaee and Ree, 2001. Barbehenn and Peter Constablel, 2011.

There is no differentiation in the market as the production and quality are similar to the mushroom growers' upwardly generalized cultivation technology. Therefore, the method to solve this difficulty is to produce a mushroom that feels the difference in functionality, efficacy, and taste with differentiated, high-quality mushrooms or to reduce production costs.

Taste and Efficacy This is a method that uses the tannin properties of chestnut yulpi as a factor that can reduce production costs Yulpi produces a large amount of by-products and cannot be used immediately as manure due to the nature of the yulpi, but has to be disposed of at a cost. Yulpi tannin ingredient was used for mushroom cultivation, and chestnut growers easily consume by-products of chestnuts, which are difficult to throw away, and mushroom growers increase productivity by reducing imported mushroom raw materials and improving the efficacy and quality of mushrooms.

In addition, the hydrolyzed tannins of chestnut yulhu dissolve in water to produce commercial products using the yulhu characteristics. However, condensed tannins are not decomposed in water, so it is difficult to use them commercially. In order to solve this problem, the condensed tannin is decomposed and absorbed using the basidiomycete, and then the basidiomycete that decomposed the condensed tannin is dissolved using distilled water or ethanol and extracted. It can be confirmed that polysaccharides are produced, and this can be used commercially.

When culturing yulpi from chestnuts in a basidiomycete mushroom medium with 1-100% of the medium raw material suitable for the characteristics of each mushroom, that is, mixing 1% or more of yulpi in the basidiomycete to be cultured, it causes a change in the physiological activity of the fungus. Therefore, if the appropriate ratio is mixed according to the characteristics of the bacteria to be cultured and the basidiomycetes are decomposed, the bacteria can be cultured for a specific use, and the vitality and mycelial density of the bacteria are improved to create a medium with the conditions for producing good mushrooms. . In particular, it creates a change in the morphology and population change of the primordial body, which is the basis for the development of mushrooms, and can obtain a higher mushroom yield compared to the existing cultivation method.

Among polyphenols in agricultural by-products, water-soluble polyphenol hydrolysis (gallotannin and ellagitannins) dissolves well in water, ethanol, etc. It is difficult to use it effectively. In particular, Yulpi contains good ingredients such as anti-cancer, skin, diabetes, etc., but its use is limited to hydrolysis, and numerous methods have been previously described for condensed tannins containing a large amount of polyphenols, but most of them are experimental methods. It is used as a powder, but its utility is limited because it is difficult to decompose condensed tannins.

Condensed (proanthocyanidin) tannins that are difficult to decompose using edible mushroom basidiomycetes are decomposed. Can be used in industry.

In general, the cultivation of commercially available mushrooms has stabilized upwards in technology and production, but it is difficult to obtain profitability due to fierce competition due to overproduction and lack of price competitiveness.

Cultivation of basidiomycetes using chestnut yulhu will reduce the cost of purchasing raw materials, increase the quality and production of mushrooms due to strong mycelial activity, and increase consumer preference by adding lightness to the taste of mushrooms.

The existing raw materials for edible mushrooms of Basidiomycetes used are sawdust, cottonseeds, and rice straw, which show general mushroom components because they have low polyphenol components. It also affects the efficacy of existing mushrooms, creating a new type of polysaccharide combined with chestnut yulpi polyphenol.

A new type of polyphenol by the decomposition of basidiomycetes of chestnut yulpi is a polysaccharide that has not been created before, and it is expected to bring about the development of a new bio-industry type as a new polyphenol material in the future.

Since tannins generally have antibacterial and polyphenol components, it is generally not applied to the production of basidiomycetes, and there are no international papers or patents on the bioactive decomposition of chestnut yulpi tannins for access to basidiomycetes.

In particular, genetic analysis of biological mushrooms, analysis of components, analysis of compounds, etc., which are being actively carried out, are actively applied to production to produce stable mushrooms. It is expected to show excellent efficacy and taste quality as a mushroom cultivation approach.

For shiitake mushrooms, mix 1%-100% of agricultural by-products containing tannins and free sugar, and more appropriately 20%-100% chestnut yarrow, or 50-100% yarrow hide, common oyster mushroom, king oyster mushroom, etc. are medium mycelium. By increasing the density and hardening of the high cell mass, the yield and quality can be increased. In particular, the aerial mycelium cell mass, which is the source of the occurrence of leaf mushrooms, is significantly increased, which is the basis for the development of high-quality leaf mushrooms.

1 shows a test photograph of mycelial elongation and activity evaluation of the basic PDA medium control yulpi-added medium for each strain.
Figure 2 shows the results of the basic test for basidiomycetes yulpi liquid culture.
Figure 3 shows the experimental results of the addition of shiitake mushroom yulpi and general sawdust medium.
Figure 4 shows the test results of the addition of oyster mushroom yulpi and general sawdust medium.
Figure 5 shows the test results of the addition of pine mushroom yulpi and general sawdust medium.
Figure 6 shows the test results of the addition of yulpi leaf mushroom and general sawdust medium.
7 shows the results of comparison of the occurrence of leaf mushrooms according to the amount of yulpi added.
8 shows the results of comparison of the yield quality of oyster mushrooms using the prepared basidiomycetes.
9 shows the results of comparison of the yield quality of leaf mushrooms using the prepared basidiomycetes.
10 shows the results of comparison of the yield quality of King oyster mushroom using the prepared basidiomycetes.
11 to 14 show photos of mycelial elongation and activity evaluation test for each strain of the basic PDA medium control yulpi-added medium for each strain.

Hereinafter, embodiments of the present invention will be described in more detail with reference to the accompanying drawings. However, the following examples are only intended to materialize the contents of the present invention, and the present invention is not limited thereto.

Polyphenols are tannins found in virtually all plant families. Although the larger polyphenols are often concentrated in leaf tissue, epidermis, bark layer, flowers and fruits, they play an important role in the decomposition and nutrient cycle of forest litter in forest ecosystems. The absolute concentrations of total phenols in plant tissues vary greatly depending on the literature source, the polyphenol type, and the assay.

With the focus on polyphenols that play an important role in decomposition and nutrient cycles in forest ecosystems, the correlation between basidiomycetes and polyphenols was confirmed, and a method of using agricultural by-products discarded around the report was derived.

Agricultural by-products with a lot of tannins in Korea are chestnut yulpi, and the consumption trend of chestnuts is in the form of peeled chestnuts processed by removing the skins of chestnuts, and export and domestic consumption are promoted. Use of tannins Livestock feed may have a temporary price advantage, but it is described in most papers that tannins interfere with protein absorption.

In general, they are reluctant to grow mushrooms because of the antibacterial component of tannin (chestnut yulpi). Therefore, there is no research literature or patent on mushroom cultivation using tannin and its free components so far.

Polyphenol tannins contained in chestnut yulpi are not suitable for culturing basidiomycetes because they are known for their antioxidant/antibacterial effects. A method for culturing basidiomycetes with

Therefore, to investigate the possibility of cultivation of basidiomycetes using chestnut yulpi tannins, the relationship between chestnut yulpi and basidiomycetes was tested.

<Basic test for basidiomycete mycelia activity of chestnut yulpi>

In order to confirm the effect of chestnut yulpi on mushroom cultivation, the following experiments were performed to measure mycelial height, mycelial density, and aerial mycelium based on the basal medium PDA. The medium composition is the same as the composition table of the basic P.D.A medium and Yulpi-added medium in Table 1, and the results are the same as the photos of mycelial elongation and activity evaluation of the basic P.D.A medium control Yulpi-added medium for each strain in FIG. 1. In addition, FIGS. 11 to 14 show pictures of mycelial elongation and activity evaluation for each strain by time of the basic P.D.A medium control yulpi-added medium for each strain.

In all basidiomycetes, the mycelial elongation rate is excellent, and the mycelium density and aerial mycelium development are improved. Therefore, chestnut yulpi has a great effect on the activity of basidiomycetes and increases the ability to decompose and elongate basidiomycetes, such as in mushroom cultivation.

Test Badge Name test medium composition Standard PDA medium composition Potato Dextrose Agar (3.9g)+ Agar (1.5g)+ Distilled Water 100ml Medium composition with oyster mushroom yulpi added Potato Dextrose Agar (3.9g)+ Agar(1.5g)+
100ml of supernatant (10g of Yulpi + 200ml of distilled water boiled for 1 hour) Medium composition with yulpi-added leaf mushroom Potato Dextrose Agar (3.9g)+ Agar(1.5g)+
100ml of supernatant (10g of Yulpi + 200ml of distilled water boiled for 1 hour) Composition of medium with shiitake mushroom yulpi Potato Dextrose Agar (3.9g)+ Agar(1.5g)+
100ml of supernatant (10g of Yulpi + 200ml of distilled water boiled for 1 hour) Medium composition with oyster yulpi added Potato Dextrose Agar (3.9g)+ Agar(1.5g)+
100ml of supernatant (10g of Yulpi + 200ml of distilled water boiled for 1 hour)

Table 1 Basic P.D.A Medium and Yulpi-Added Medium Composition Table

As can be seen from the results of Fig. 1 and Figs. 11 to 14, when each mushroom strain was inoculated and cultured for the same time, the mycelial growth was excellent in the medium with chestnut yulpi than the basic P.D.A medium, and this mycelial growth Compared to the existing mushroom cultivation, it becomes a matrix that can make excellent mushrooms and is a condition that can cause various compositional changes.

<Liquid culture test of basidiomycetes chestnut yulpi>

Next, we tested whether or not Yulpi can be used for inoculation of basidiomycetes for mushroom cultivation or a liquid culture method capable of extracting basidiomycetes components by rapidly culturing basidiomycetes.

Test liquid culture medium Medium name test medium composition Composition of the reference solution culture medium (distilled water 1000ml + potato 200g) ⇒ supernatant after boiling Composition of oyster mushroom yulpi-added liquid culture medium (distilled water 1000ml + potato 200g + yulpi 50g)⇒ supernatant after boiling Composition of medium for culturing with yulpi-added leaf mushroom (distilled water 1000ml + potato 200g + yulpi 50g)⇒ supernatant after boiling Composition of shiitake mushroom yulpi-added liquid culture medium (distilled water 1000ml + potato 200g + yulpi 50g)⇒ supernatant after boiling Composition of culture medium with oyster yulpi added solution (distilled water 1000ml + potato 200g + yulpi 50g)⇒ supernatant after boiling

Table 2 Composition Table of Common Potato Medium and Yulpi Potato Medium

The test results are shown in FIG. 2 . The normal potato medium for oyster mushrooms took 22 days to incubate, but the yulpi potato medium is as fast as 17 days. The culturing period for shiitake mushroom and potato medium is about 30 days, but for Yulpi potato medium, it is as fast as 23 days. The normal potato medium for king oyster mushroom took 23 days to incubate, but the yulpi potato medium is as fast as 19 days.

<Comparative test of basidiomycete chestnut yulpi general sawdust culture>

As test samples, oak sawdust (a sawdust manufacturing plant in Eunsan, Buyeo), sawdust from mulch (sawdust in Namwon near Mt. Jirisan), chestnut husks (a waste from the forest association of Buyeo-gun), bran and soybean meal were purchased from Pyeongpack LOHAS Co., Ltd. and used.

<Preparation for basic experiment>

Strain used: The test was conducted after confirming that it was a stable strain free from contamination by isolating the self-owned shiitake mushroom strain, oyster Hwaseong No. 5 of Korea Wongyun, and shiitake mushroom of Okcheon.

Cultivation: Cultivation was carried out at 70% humidity (10% deviation) by using a humidifier in an incubator at normal room temperature (a room set at 24 degrees with an automatic heating device). In order to confirm this, the representative mushrooms of basidiomycetes, oyster mushroom, shiitake mushroom, and king oyster mushroom, were tested.

<shiitake mushrooms>

The experimental results of the addition of shiitake mushroom chestnut yulpi and general sawdust medium are shown in FIG. 3 .

The left side of FIG. 3 is a commonly used shiitake mushroom medium (oak (70%) + bran (15%) + soybean meal (15%)), and the right medium is a medium with chestnut yarrow (oak (30%) + bran (15)). %) + soybean meal (15 %) + yulhide (40%)). The mycelial elongation rate and the density form of the bacteria were compared according to the media with different contents.

Just as the shape, taste, and shape are different depending on the type of basidiomycete, the shape and shape of the shiitake mushroom strain according to the polyphenol type of agricultural by-products are also different. Although it is known that the medium to which Yulpi is added cannot cultivate mushrooms due to the commonly known polyphenol component, the mycelial elongation rate is slightly slow, but the mycelium density and the mycelium density that constitutes reproductive growth for mushroom development is high.

<Oyster mushroom>

4 shows the results of the addition of oyster mushroom chestnut yulpi and general sawdust medium.

Compared to the left standard medium (oak (70%) + bran (15%) + soybean meal (15%)) for oyster mushroom, the mycelial growth rate and mycelial density are not affected by chestnut yarrow, and there is a slight difference and similar appearance with the naked eye. , The medium with chestnut yarrow on the right side (oak (30%) + bran (15%) + soybean meal (15%) + yarrow (40%)) affects the occurrence, growth and quality of mushrooms and is the main cause of productivity. Able to know.

common badge oyster Yulpi Badger Tari the occurrence of mushrooms Mushroom occurrence distribution is high Mushroom occurrence distribution is low growth Growth by large numbers of individuals Growth in small numbers Quality (Marketability) upper middle lower upper middle lower Economics upper middle lower upper middle lower

<Leaf Mushroom>

Fig. 6 shows the results of the test results of the addition of chestnut chestnut mushroom and general sawdust medium.

The mycelial growth rate of shiitake mushrooms showed that the left conventional medium (oak (70%) + bran (15%) + soybean meal (15%)) showed excellent oak bran and soybean meal medium, and the medium added with polyphenol medium of agricultural by-products (oak (oak) 30%) + bran (15%) + soybean meal (15%) + yulhide (40%)) seem to have lower elongation speed and density. In terms of mycelial growth rate, the left test tube occupies and elongates faster, but the right test tube looks better in overall mycelial density.

Hyphal density and the development of aerial hyphae show differences according to the content of chestnut yarrow. Aerial hyphae and mycelial density are the basis of fruiting body formation, and there is a big difference in the quality and yield of mushrooms.

Common Badge Mushroom Yulpi Medium Mushroom the occurrence of mushrooms Strain selective development by gene analysis expression pattern A common medium cultured with Yulpi medium is the overall development growth universal growth universal growth Quality (Marketability) award special prize taste General taste with fishy fishy taste Taste that leaves clean and clean inside

<Comparative test for yield quality of basidiomycetes chestnut yulpi common sawdust mushroom>

< Oyster mushroom >

The cultivation method of oyster mushroom using chestnut yulpi is shown in Table 5 below.

Cultivation method General Sawdust Mushroom Medium Yulpi Mushroom Badge medium composition Sawdust 70% + bran 15% + soybean meal 15% Chestnut skin 70% + bran 15% + soybean meal 15% Media weight 2.8 kg moisture 65% 2.8 kg moisture 65% culture conditions Humidity 70%, Temperature 23 ±1 ℃ Humidity 70%, Temperature 23 ±1 ℃ incubation day 25 days 21 days occurrence condition Temperature 17℃, humidity 80% Temperature 17℃, humidity 80% growing harvest conditions Temperature 18℃, Humidity 80% Temperature 18℃, Humidity 80%

A comparison of the yield quality of oyster mushrooms using the prepared basidiomycetes is shown in FIG. 8 .

When comparing the yield and quality of the basidiomycete oyster mushroom in Fig. 8, the overall quality of oyster mushroom is higher than that of the general sawdust medium (sawdust 70% + bran 15% + soybean meal 15%). ) of oyster mushroom was excellent, and the yield of general sawdust medium was 350g and Yulpi medium was 480g.

<Leaf Mushroom>

The method of cultivation of shiitake mushrooms using chestnut yulpi is shown in Table 6 below.

Cultivation method General commercial mushroom medium Yulpi Mushroom Badge medium composition Sawdust 70% + bran 15% + soybean meal 15% Chestnut skin 70% + bran 15% + soybean meal 15% Media weight 2.8 kg moisture 65% 2.8 kg moisture 65% culture conditions Humidity 70%, Temperature 23 ±1 ℃ Humidity 70%, Temperature 23 ±1 ℃ incubation day 51 days 45 days occurrence condition Temperature 17℃, humidity 80% Temperature 17℃, humidity 80% growing harvest conditions Temperature 18℃, Humidity 95% Temperature 18℃, Humidity 95%

9 shows a comparison of the yield quality of leaf mushrooms using the prepared basidiomycetes.

The quality of the mushroom medium is similar in appearance, but the taste is that the Yulpi medium removes the fishy taste of the general sawdust medium, and you can feel the protein and clean taste. As a result, Yulpi medium was found to be excellent.

<Oyster mushroom>

The cultivation method of king oyster mushroom using chestnut yulpi is shown in Table 7 below.

Cultivation method General commercial mushroom medium Yulpi Mushroom Badge medium composition Sawdust 70% + bran 15% + soybean meal 15% Chestnut skin 70% + bran 15% + soybean meal 15% Media weight 2.8 kg moisture 65% 2.8 kg moisture 65% culture conditions Humidity 70%, Temperature 23 ±1 ℃ Humidity 70%, Temperature 23 ±1 ℃ incubation day 30 days 27 days occurrence condition Temperature 17℃, humidity 80% Temperature 17℃, humidity 80% growing harvest conditions Temperature 18℃, Humidity 95% Temperature 18℃, Humidity 95%

A comparison of the yield and quality of pine mushrooms using the prepared basidiomycetes is shown in FIG. 10 .

When comparing the yield and quality of basidiomycetes cypress mushroom in Fig. 10, the overall quality of King oyster mushroom is higher than that of general sawdust medium (sawdust 70% + bran 15% + soybean meal 15%). ) of king oyster mushroom was excellent, and the yield of general sawdust medium was 550g and Yulpi medium was 730g.

<Comparative test for composition analysis of basidiomycetes chestnut yulpi common sawdust mushroom>

1. General component analysis result

division moisture views george Crude Protein* carbohydrate Mushroom Sawdust Cultivation 9.88 7.76 3.55 29.03 49.78 Mushroom yulpi cultivation 7.12 5.94 1.80 25.40 59.74

Crude protein: Damas method 2. Minerals (mg/100g)

division Ca Fe K Mg Mn Na P Zn Mushroom Sawdust Cultivation 4.60 2.61 1,007.19 118.53 0.68 11.32 787.35 7.24 Mushroom yulpi cultivation 18.30 2.62 709.02 37.53 0.68 26.08 769.80 7.45

As a result of the component analysis, it can be seen that Yulpi-medium mushroom shows an increase in the carbohydrate portion containing β-gulucan, which is the crystallization of mushroom efficacy, and in particular, the Ca ion of the protease protease is increased more than 4 times.

Since each protease has specificity, it has the property of easily cleaving the peptide bond at the amino or carboxy side of a specific amino acid residue in a protein.

Proteases are used when ingesting amino acids, a material for making proteins, by digesting proteins in food, on the contrary, when removing unnecessary proteins inside and outside cells, or in precursors of proteins and peptides with various biological activities, including enzymes. It plays an important role when activating and separating unnecessary parts.

Examples include pepsin from gastric juice, trypsin, chymotrypsin, and elastase from pancreatic juice, various cathepsins present in lysosomes in intracellular organelles, and thrombin that converts fibrinogen into fibrin.

Claims (8)

A method for culturing basidiomycetes, characterized in that using chestnut yulpi. The method of claim 1,
The basidiomycete culture method, wherein the basidiomycete is at least one selected from shiitake mushroom, oyster mushroom, king oyster mushroom and shiitake mushroom. The method of claim 1,
The culturing method includes inoculating and culturing basidiomycetes in a medium containing Yulpi.
Including, basidiomycetes culture method. How to decompose chestnut yulpi using basidiomycetes. Edible mushroom thickening material containing chestnut yulpi. A method of increasing the growth rate of basidiomycete culture using chestnut yulpi. A medium for culturing basidiomycetes containing chestnut yulpi. 8. The method of claim 7,
The yulpi is a medium for culturing basidiomycetes that contains 1 to 100 parts by weight based on 100 parts by weight of the medium.

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