KR20210027873A - Culture method for enhancing content of Paecilomyces javanicus mycelium using Light-Emitting Diode - Google Patents

Abstract

The present invention relates to a culture method for increasing the content of a Paecilomyces javanicus mycelium using an LED, wherein the culture method thereof comprises: an inoculation step of inoculating a medium with a Paecilomyces javanicus mycelium; and a culturing step of culturing the inoculated medium under light conditions to obtain a Paecilomyces javanicus mycelium. It has been confirmed that the maximum amount of growth of Paecilomyces javanicus mycelium is shown depending on the medium and light conditions. Through the reaction surface analysis, the optimal amount of light, the glucose content of the medium, and the culture time conditions are derived, and the maximum growth amount of the Paecilomyces javanicus mycelium can be efficiently obtained by using the same as the culture conditions.

Description

Culture method for enhancing content of Paecilomyces javanicus mycelium using Light-Emitting Diode}

The present invention relates to a cultivation method for increasing the content of the mycelium of Cordyceps sinensis using LED.

Cordyceps hacho (冬蟲夏草) was named because it comes out as a bug in winter and a mushroom in summer. About 800 species are known as most of them as entopathogenic fungi, and it is characterized by being infected with host insects and producing a fruiting body by growing mycelium inside. These cordyceps have been used as medicinal herbs in Asia (China, Korea, Japan, etc.) from a long time ago. For its efficacy, it has been used as a good medicine for enhancing immunity, preventing adult diseases, asthma, and anemia.

In addition, cordyceps sinensis belongs to the group with the highest protein content (more than 28% of dry content) among crops, and contains a large amount of substances that enhance immunity. In particular, cordycepin (3'-deoxy-adenosine), which is found to be an isomer of quinic acid, is included among the immunologically active substances of Cordyceps sinensis. Cordycepin is a nucleic acid substance that is involved in the genetic information of cells and activates a reduced immune function to prevent normal cells from being converted into cancer cells. In a clinical trial with cordyceps sinensis, it was confirmed that the content of NK cells (natural killer cells), which are immune cells that kill cancer cells, and cytokines secreted from immune cells, increased by 18% to 25%. It is utilized and is receiving a lot of attention as a health functional food.

Based on these results, cordycepin-containing cordyceps has been certified by the Ministry of Food and Drug Safety as a health functional food in Korea, and various functions such as antiviral and anti-inflammatory effects in addition to immunity enhancing activity and anticancer activity have been announced.

Due to the above functions, cordyceps are consumed a lot among mushrooms, but the amount collected from nature is very small, so it cannot meet the demand, so many farmers around the world have made cultivation houses using artificial cultivation methods and cultivated them in large quantities. However, cordyceps is a plant that cannot be sprayed with pesticides because its growth rate is slow and sensitive to the external environment, so cultivation is very difficult. In addition, although it has succeeded in mass cultivation, it is known that the yield rate of cultivation of cordyceps is low even in farms that have been cultivated for a long time. That is, when 10 cells are cultured and harvested, about 1 to 2 cells are contaminated or abnormal strains grow, resulting in a lower harvest rate.

Recently, research on the production of mycelium rather than the production of fruiting bodies of Cordyceps sinensis is attracting attention, and the mycelium has the advantage of being able to cultivate for a short period of time than fruiting bodies and that it can be cultivated in a small place at low cost. In addition, Feng et al . (2018) and Tang et al . According to (2018), it was reported that there was little difference in components between fruiting bodies and mycelium of Cordyceps sinensis. Therefore, it is considered economically advantageous to cultivate mycelium that can be mass-produced in a short period of time rather than fruiting bodies. However, until now, studies on optimal conditions suitable for increasing the production of cordyceps mycelium are insufficient.

Therefore, there is a need for research on an optimal culture method for increasing the growth amount of mycelium of Cordyceps sinensis containing cordycepine.

1. Korean Patent Registration No. 10-0465283 (2003.08.14.)

It is an object of the present invention to provide an optimal method for culturing a mycelium of Snow Cordyceps sinensis for the maximum growth amount of Cordyceps sinensis.

In addition, another object of the present invention is to provide a mycelium of Cordyceps sinensis according to the above culture method.

In addition, another object of the present invention is to provide a method for predicting the maximum growth amount of mycelium Cordyceps sinensis.

In order to achieve the above object, the present invention is an inoculation step of inoculating the mycelium of Cordyceps sinensis into a medium; And a culturing step of culturing the inoculated medium under light conditions to obtain a mycelium of Cordyceps sinensis; It provides a cultivation method for increasing the content of the mycelium cordyceps sinensis comprising a.

In addition, the present invention provides a mycelium cultivated in accordance with the method of culturing the mycelium of Cordyceps sinensis.

In addition, the present invention in the method for predicting the dry weight of the mycelium of Cordyceps sinensis, (a) the amount of light (X ₁ ) by the Box-Behnken design, the amount of glucose (glucose) of the medium (X ₂ ), culture Designing an experimental range by coding as -1, 0, and 1 for time (X _{3 );} (b) obtaining experimental values for the amount of light, the glucose content of the medium, and the incubation time in the experimental range designed in step (a); (c) deriving a quadratic regression model represented by Equation 1 below using the experimental value of step (b); And (d) predicting the dry weight of the mycelium of Cordyceps sinensis by analyzing the quadratic regression model represented by Equation 1 derived in step (c) by variance analysis (ANOVA). It provides a method for predicting the dry weight of the mycelium Cordyceps sinensis, comprising a.

[Equation 1]

Y ₁ =14.53-1.14X ₁ -0.9713X ₂ +0.5025X ₃ -0.9925X ₁ X ₂ +0.5450X ₁ X ₃ +0.0100X ₂ X ₃ +2.65X ₁ ² +2.19X ₂ ² +2.58X ₃ ²

(In Equation 1, Y ₁ is the dry weight (g/L) of the mycelium of Cordyceps sinensis, X ₁ is the amount of light (code unit), X ₂ is the glucose content of the medium (code unit), and X ₃ is the culture time (code unit). Means.)

According to the present invention, when cultivated by irradiating green LED light in a PDB (potato dextrose broth) liquid medium, the snowflake cordyceps mycelium showed the maximum growth amount, and the optimal amount of light, the glucose content of the medium, and the culture time conditions were determined through reaction surface analysis. Derived and used as a culture condition, there is an effect of efficiently obtaining the maximum growth amount of the mycelium of Cordyceps sinensis.

In addition, the mycelium of Cordyceps sinensis obtained according to the cultivation method according to the present invention contains cordicepin, which has excellent immune enhancing activity, anticancer activity, antiviral and anti-inflammatory effects, and thus can be usefully used as a health functional food.

1 is a snowflake cordyceps ( Paecilomyces) javanicus ) is a diagram showing the mycelium.
Figure 2 is a view showing the growth of mycelium according to the type of liquid medium of Cordyceps sinensis (data shows the results of three experiments as mean ± standard deviation).
Figure 3 is a view showing the growth of mycelium according to the culture period of Cordyceps sinensis (data shows the results of three experiments as mean ± standard deviation).
4 is a diagram showing a shaking incubator in which a fluorescent lamp and UV-A are installed.
5 is a view showing a fluorescent lamp and a control box for adjusting the amount of UV-A light and on/off.
6 is a view showing the change in mycelium growth of Snow Cordyceps sinensis according to irradiation with a fluorescent lamp and UV-A (data are shown as mean ± standard deviation of the results of three experiments).
7 is a diagram showing a shaking incubator in which single LED lights (red, green, blue) are installed.
8 is a view showing a control box for adjusting the amount of light and on/off of a single LED light.
9 is a view showing the change in mycelium growth amount according to the LED single light (red, green, blue) irradiation of the Snow Cordyceps sinensis (data shows the results of three experiments as mean ± standard deviation).
Figure 10 is a diagram showing the change in the growth amount of the mycelium of Cordyceps sinensis according to the irradiation time of LED green light (data shows the results of three experiments as mean ± standard deviation).
11 is a view showing the change in mycelium growth according to the type of LED single light and mixed light of the Snow Cordyceps sinensis (data shows the results of three experiments as mean ± standard deviation).
12 is a view showing the change in mycelium growth amount according to the irradiation time of mixed light red*green in the mycelium of Cordyceps sinensis (data shows the results of three experiments as mean ± standard deviation).
13 is a view showing the correlation between the mycelium culture conditions and mycelium growth amount.
14 is a view showing a fit model between the LED control conditions and the mycelium growth amount of Cordyceps sinensis.
15 is a view showing the optimal LED control conditions for the maximum amount of mycelium of the mycelium of Snow Cordyceps sinensis.
16 is a view showing the suitability of the optimal LED control condition response model for the maximum amount of mycelium of the mycelium of Cordyceps sinensis.

Hereinafter, the present invention will be described in detail.

The present inventors confirmed that when the green LED light was irradiated for 4 to 5 days at 12 hours per day in a liquid medium of PDB (potato dextrose broth) and cultured, it showed the maximum growth amount of the mycelium Cordyceps sinensis, and is optimal through reaction surface analysis. The present invention was completed by finding that the maximum amount of growth of the mycelium of Cordyceps sinensis can be efficiently obtained by deriving the light quantity, the glucose content of the medium, and the culture time conditions, and using this as a culture condition.

The present invention is an inoculation step of inoculating the mycelium of Cordyceps sinensis into the medium; And a culturing step of culturing the inoculated medium under light conditions to obtain a mycelium of Cordyceps sinensis; It provides a cultivation method for increasing the content of the mycelium cordyceps sinensis comprising a.

At this time, the medium may be a potato dextrose broth medium, and the cultivation may be performed for 3 to 6 days, preferably 4 to 5 days.

In addition, the light condition is to cultivate the mycelium by irradiating the LED single light or mixed light onto the inoculated medium, and the LED single light is green light having a wavelength of 500 nm to 550 nm, and the LED mixed light is green light having a wavelength of 500 nm to 550 nm and It is characterized in that the light is a mixture of red light having a wavelength of 620 nm to 780 nm, and preferably, LED green light having a wavelength of 500 nm to 550 nm may be used, and at this time, it may represent the maximum growth amount of the mycelium of Cordyceps sinensis.

In addition, it is characterized in that the mycelium is cultured by irradiating the LED single light or the mixed light on the inoculated medium for 5 to 15 hours per day, and preferably, it may be irradiated for 12 hours per day, but is not limited thereto. .

At this time, if the cultivation conditions and light conditions as described above are deviated, the mycelium of Snow Cordyceps sinensis according to the present invention may not be properly obtained, or the yield of Snow Cordyceps sinensis may be significantly lowered, resulting in a problem that is not economical.

In addition, the dry weight of the mycelium of Cordyceps sinensis may be characterized in that it has a correlation with the amount of light, the glucose content of the medium, and the culture time as shown in Equation 1 below.

[Equation 1]

Y ₁ =14.53-1.14X ₁ -0.9713X ₂ +0.5025X ₃ -0.9925X ₁ X ₂ +0.5450X ₁ X ₃ +0.0100X ₂ X ₃ +2.65X ₁ ² +2.19X ₂ ² +2.58X ₃ ²

(In Equation 1, Y ₁ is the dry weight (g/L) of the mycelium of Cordyceps sinensis, X ₁ is the amount of light, X ₂ is the glucose content of the medium, and X ₃ is the culture time.)

At this time, the cultivation conditions of the amount of light, the glucose content of the medium, and the cultivation time for obtaining the maximum dry weight of the mycelium Cordyceps sinensis are characterized in that, respectively, 1000 to 1500 lux, 1 to 10 g/50 mL, and 80 to 120 hours, preferably Culture conditions of the amount of light, the glucose content of the medium, and the culture time may be 1335.16 lux, 5.01181 g/50mL, and 97.6614 hours, respectively, but are not limited thereto.

In addition, the present invention provides a mycelium of Snow Cordyceps sinensis cultured according to the method of culturing Snow Cordyceps sinensis mycelium.

Since the mycelium of Cordyceps sinensis cultivated as described above contains cordycepin, it may be used as a health functional food for immune enhancing activity and anticancer activity.

In addition, the present invention in the method for predicting the dry weight of the mycelium of Cordyceps sinensis, (a) the amount of light (X ₁ ) by the Box-Behnken design, the amount of glucose (glucose) of the medium (X ₂ ), culture Designing an experimental range by coding as -1, 0, and 1 for time (X _{3 );} (b) obtaining experimental values for the amount of light, the glucose content of the medium, and the incubation time in the experimental range designed in step (a); (c) deriving a quadratic regression model represented by Equation 1 below using the experimental value of step (b); And (d) predicting the dry weight of the mycelium of Cordyceps sinensis by analyzing the quadratic regression model represented by Equation 1 derived in step (c) by variance analysis (ANOVA). It provides a method for predicting the dry weight of the mycelium Cordyceps sinensis, comprising a.

[Equation 1]

Y_One=14.53-1.14X_One-0.9713X₂+0.5025X₃-0.9925X_OneX₂+0.5450X_OneX₃ +0.0100X₂X₃+2.65X_One ²+2.19X₂ ²+2.58X₃ ²

(In Equation 1, Y ₁ is the dry weight (g/L) of the mycelium of Cordyceps sinensis, X ₁ is the amount of light (code unit), X ₂ is the glucose content of the medium (code unit), and X ₃ is the culture time (code unit). Means.)

At this time, the cultivation conditions of the amount of light, the glucose content of the medium, and the cultivation time for obtaining the maximum dry weight of the mycelium Cordyceps sinensis are characterized in that, respectively, 1000 to 1500 lux, 1 to 10 g/50 mL, and 80 to 120 hours, preferably Culture conditions of the amount of light, the glucose content of the medium, and the culture time may be 1335.16 lux, 5.01181 g/50mL, and 97.6614 hours, respectively, but are not limited thereto.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for describing the present invention in more detail, and that the scope of the present invention is not limited by these examples according to the gist of the present invention, to those of ordinary skill in the art to which the present invention pertains. It will be self-evident.

< Example 1> Establishment of liquid culture technology for cordyceps mycelium

1. Disclosure material

In order to evaluate the growth amount of cordyceps mycelium, snowflake cordyceps ( Paecilomyces javanicus ) (KCCM 60299) was pre-sale from the Korea Microbial Conservation Center (KCCM) (Fig. 1).

2. Snowflake Cordyceps ( Paecilomyces javanicus ) Liquid culture and growth analysis

Snowflake Cordyceps ( Paecilomyces javanicus ) of three kinds (YMG (yeast extract, malt extract, glucose medium), PDB (potato dextrose broth), SDB (Sabouraud dextrose broth)) having the culture composition shown in Table 1 below to establish the culture environment conditions for increasing the growth amount of A liquid culture medium was used.

Badge name Medium pH Medium composition (g/L) Yeast extract Malt extract Glucose Potato extract Peptone YMG 6.0 4 10 4 - - PDB 5.2 - - 20 4 - SDB 4.5 - - 20 - 10

Snow flower cordyceps (Paecilomyces) sold from the KCCM javanicus ) mycelium was cultured for 7 days at 25°C in an incubator using a PDA (Potato Dextrose Agar) plate medium. Thereafter, the edges of the cultured mycelium were inoculated into liquid medium (YMG, PDB, SDB) by 3 cores using a cork borer with a diameter of 5 mm, and cultured with shaking (100 rpm) at 25° C. for 7 days to produce a liquid seed. The amount of mycelium growth of the liquid seed cultured at intervals was measured.

The YMG, PDB, or SDB medium was added to each of 50 mL in a 100 mL Erlenmeyer flask, sterilized at 121° C. for 15 minutes, and then cooled and used.

In addition, the growth amount of the cultured snowflake cordyceps mycelium was determined by Carvajal et al . It was measured by the following method by the dry weight measurement method of (2012).

① Transfer the cultured mycelium culture medium to a falcon tube.

② Centrifugation (4000 rpm, 10 min, 2 times, Union 32R Plus, HANIL, Korea) to separate the culture medium and mycelium.

③ Dry the falcon tube contained in the mycelium at 60℃ for 24 hours.

④ Derive mycelium weight by using the total weight of the falcon tube measured in advance.

3. Snowflake Cordyceps ( Paecilomyces javanicus ) According to the liquid culture medium

The growth amount of mycelium according to the type of liquid medium of the mycelium cordyceps sinensis is shown in FIG. 2.

As a result, it was confirmed that the growth amount of the mycelium of Cordyceps sinensis was 10.2 g/L when cultured in PDB (Potato Dextros Broth) medium, showing the highest growth amount.

In addition, as a result of analyzing the growth amount of Cordyceps sinensis according to the cultivation period, as shown in FIG. 3, the growth amount of mycelium after 5 days was indicated, and the culture period of the mycelium of Cordyceps sinensis was fixed to 5 days.

< Example 2> Evaluation of the effect of fluorescent lamps and UV-A on the mycelium growth of Cordyceps sinensis

1. Disclosure material and method of culturing mycelium of Cordyceps sinensis

A fluorescent lamp and UV-A (356 nm) were installed in the shaking incubator as shown in FIG. 4 to evaluate the change in the growth amount of the mycelium of Cordyceps sinensis sold in <Example 1-1> according to the irradiation of a fluorescent lamp and UV-A. The light source was installed at a height of about 30cm of the culture solution, and the luminous intensity was set to ^{100pmol·m -2} ·s ^-1.

In addition, the control box for the on/off of the light source was purchased and installed from Bitsol LED, an LED lighting brand for plant growth of ESREDS, as shown in FIG. 5, and the cultivation was performed as in <Example 1-2>. As conditions, liquid seed was produced by shaking culture (100 rpm) for 7 days at 25° C. in PDB medium, and fluorescent lamp and UV-A were irradiated for 5 days at 12 h/day, and the growth amount was measured by a dry weight measurement method.

As a control, the mycelium of Cordyceps sinensis cultivated under dark culture conditions was used.

2. Growth amount according to fluorescent lamp and UV-A light source of Snowflake Cordyceps

6 shows the growth amount of mycelium according to the fluorescent lamp and UV-A irradiation of the Snow Cordyceps sinensis, and it was confirmed that the Snow Cordyceps had the largest growth amount of mycelium when cultured in a fluorescent light source than in dark culture or UV-A, and than in dark culture. It was found that about 2 times more mycelium was produced.

< Example 3> LED Monolithic Evaluation of mycelium growth of Cordyceps sinensis according to control conditions (wavelength type, irradiation time)

1. Disclosure material and method of culturing mycelium of Cordyceps sinensis

In order to evaluate the change in mycelium growth of the Snow Cordyceps sinensis according to the irradiation of the LED single light, a single LED light was installed in the shaking incubator as shown in FIG. 7. For single light, a total of 3 colors were installed: red, green, and blue, and 120 pieces were installed for each color (Table 2). The light source was installed at a height of about 30cm of the culture solution, and the luminous intensity was set to 64.9-108.0 pmol·m ^-2 ·s ^-1 .

In addition, the LED and the control box were installed by purchasing the LED lighting brand for plant growth of ESrez Co., Ltd., as shown in Fig. A liquid seed was produced by shaking culture for 7 days (100 rpm), and the three types of single light were irradiated for 5 days at 12 h/day and the growth amount was measured by dry weight measurement method.

As a control, cordyceps mycelium cultured under dark culture, fluorescent lamp and UV-A conditions (Example 2) was used.

color Wavelength(nm) LED quantity PCB quantity Number of LEDs per PCB Red 650 120 10 12 Green 525 120 10 12 Blue 450 120 10 12

2. LED of Snowflake Cordyceps Single light Growth according to conditions

9 shows the growth amount of mycelium according to the LED single light irradiation of the Snow Cordyceps sinensis, and it was confirmed that the Snow Cordyceps had the largest growth amount of the mycelium when irradiated with green light, which was 1.17 times greater than dark culture and 1.28 times greater than fluorescent lamps. , It showed 1.40 times higher mycelium growth than UV-A.

Therefore, LED green light is judged to be effective in increasing the growth of the mycelium of Cordyceps sinensis, and FIG. 10 shows the change in the growth of mycelium according to the irradiation time of green light among LED single lights. It was confirmed that the highest growth rate was indicated by /L.

< Example 4> LED Mixed mania Control condition ( Mixed light Evaluation of cordyceps mycelium growth according to type and irradiation time)

1. Disclosure material and method of culturing mycelium of Cordyceps sinensis

Red*green (650+525nm), red*blue (650+450nm), and green*blue (525+450nm) were used as the mixed light in order to evaluate the change in mycelium growth of the Snow Cordyceps sinensis according to the irradiation of the LED mixed light. the brightness is ^{64.9 ~ 108.0 pmol · m -2 ·} s - was set to ^one.

In addition, culture was carried out with shaking culture (100 rpm) at 25°C for 7 days in PDB medium under the same conditions as in <Example 1-2> to produce liquid seed, and 3 kinds of mixed light were irradiated for 5 days at 12 h/day. Then, the growth amount was measured by the dry weight measurement method.

2. LED of Snowflake Cordyceps Mixed light Growth according to conditions

11 shows the growth amount of mycelium according to the LED mixed light irradiation of Snowflake Cordyceps sinensis, 8.7 g/L, 7.7 g/L, and 7.9 g when irradiated with red*green, red*blue, and green*blue mixed light, respectively /L.

Therefore, when red*green was irradiated on the mycelium of Cordyceps sinensis, it was found to be most effective in increasing the growth of mycelium.

However, as in <Example 3>, when compared with green light (P. javanicus : 10.8 g/L), which was the single light that was most effective in increasing the growth of the mycelium of Cordyceps sinensis, the single light increased the growth of the mycelium of Cordyceps sinensis than the mixed light. It has been found to be effective.

Therefore, the application of LED for increasing the growth amount of the mycelium of Snow Cordyceps sinensis is judged to be more effective than the mixed light, and FIG. 12 shows the change in the growth amount of the mycelium according to the irradiation time of the red*green mixed light of the Snow Cordyceps sinensis, 6 h/day. When irradiated, it showed the highest mycelium growth.

< Example 5> Derivation of optimal control conditions of LED for maximum growth of mycelium

1. LED control conditions and mycelium Between growth Derivation of correlation

Pearson correlation coefficient (Hauke and Kossowski, 2001) and heat map (Wilf et al., 2016) were used to derive the correlation between the mycelium culture conditions of Cordyceps sinensis including LED control conditions and the growth of mycelium, and the statistical program was R program (version 3.4). .3) was used.

The parameters of the culture conditions for mycelium of Cordyceps sinensis were set as LED wavelength, LED light intensity, LED irradiation time, glucose content of the medium, and pH of the medium.

In the Pearson correlation coefficient, the higher the coefficient was, the higher the correlation between the culture condition factor and the growth amount was.In order to derive the regression equation between the LED control condition and the growth amount of cordyceps mycelium, the fit of the CurveExpert program (version 3.4.3). model was used.

As a result, the correlation between the mycelium culture conditions of Cordyceps sinensis and mycelium growth amount is shown in FIG. It was found to show a positive correlation (0.40) with mycelium growth.

In addition, the fit model between the LED control condition and the growth amount of cordyceps mycelium is shown in FIG. 14, and the LED wavelength, which had the highest correlation between the LED control condition and the mycelium growth amount, was confirmed to be the'Sinusoidal' model. The regression equation using this is shown in Table 3, and it is judged that the growth amount can be predicted using only the LED control conditions (wavelength, light amount, irradiation time) through the regression equation between the derived LED control conditions and the mycelium growth amount.

No. χ y Regression a constant One wavelength Mycelium growth y=a+bcos(cχ+d) a=130.16; b=61.38; c=2.01; d=3.11 2 Light intensity Mycelium growth y=a+bcos(cχ+d) a=10.53; b=51.98; c=2.11; d=3.17 3 Investigation time Mycelium growth y=a+bχ+cχ ² +d a=-0.355519; b=0.305731; c=-0.001638; d=0.000003

2. Establishment of optimal LED conditions for increasing mycelium growth

The optimal LED control conditions for increasing mycelium growth of Cordyceps sinensis were using the Box-Benkhen design (BBD) experimental design of the Response Surface Model (RSM), and the program was Design-Expert Software Version 10. I did.

The fixation factor was the mycelium of Snow Cordyceps (P. javanicus ), and the variables were set as the light quantity condition, the glucose content of the medium, and the cultivation time. At this time, the range of the variable was set through a preliminary experiment, and the BBD experimental design was set to a total of 17 conditions as shown in Table 4 below.

Run Independent variables
(coded) Independent variables
(actual) Mycelium dry weight (Mycelia dry
weight), g/L X _One X ₂ X ₃ X _One X ₂ X ₃ Y _One One One 0 -One 1338 7.5 24 17.83 2 0 0 0 669 7.5 72 14.02 3 One 0 One 1338 7.5 120 20.22 4 -One One 0 0 10 72 21.21 5 0 0 0 669 7.5 72 15.39 6 0 -One -One 669 5 24 20.18 7 -One 0 -One 0 7.5 24 20.46 8 One -One 0 1338 5 72 19.53 9 -One 0 One 0 7.5 120 20.55 10 0 -One One 669 5 120 20.93 11 One One 0 1338 10 72 16.16 12 0 One One 669 10 120 18.45 13 0 0 0 669 7.5 72 13.12 14 0 0 0 669 7.5 72 16.02 15 -One -One 0 0 5 72 20.61 16 0 One -One 669 10 24 17.66 17 0 0 0 669 7.5 72 14.09 Independent variables Levels -One 0 One X ₁ : Light quantity, lux 0 669 1338 X ₂ : glucose content, g/50 mL 5 7.5 10 X ₃ : Incubation time, h 24 72 120

FIG. 15 shows a three-dimensional response surface graph showing the relationship between each factor as a response surface and an induced quadratic polynomial regression model. As a result, the optimal culture condition for the maximum mycelium amount of the mycelium of Cordyceps sinensis is the amount of light of 1335.16 lux , When the glucose content of the medium was 5.01181 g/50mL, and the culture period was 97.6614 hours, it was confirmed that the mycelium growth amount was increased to 21.4606 g/L.

In addition, the second-order polynomial regression model was subjected to variate analysis (ANOVA ^{) to obtain R 2} (coefficient of determination) indicating the fitness of the reaction model as shown in FIG. 16. As a result, R ² (coefficient of determination) showed high reliability as 0.8854, and accordingly, the second-order polynomial equation derived in FIG. 15 was judged to be suitable for predicting the response value of the optimal culture conditions for the maximum mycelium amount of the Snow Cordyceps sinensis. .

Claims (14)

Inoculation step of inoculating the mycelium of Cordyceps sinensis into the medium; And
A culturing step of culturing the inoculated medium under light conditions to obtain a mycelium of Cordyceps sinensis; Cultivation method for increasing the content of the mycelium cordyceps sinensis comprising a. The method of claim 1,
The culture method for increasing the content of mycelium Cordyceps sinensis, characterized in that the medium is a potato dextrose broth medium. The method of claim 1,
Culture method for increasing the content of the mycelium Cordyceps sinensis, characterized in that the cultivation is carried out for 3 to 6 days. The method of claim 1,
The light condition is a culture method for increasing the content of the mycelium of Cordyceps sinensis, characterized in that the mycelium is cultured by irradiating the inoculated medium with LED single light or mixed light. The method of claim 4,
Single LED light is green light having a wavelength of 500 nm to 550 nm, and the mixed LED light is light obtained by mixing green light having a wavelength of 500 nm to 550 nm and red light having a wavelength of 620 nm to 780 nm. How to cultivate. The method of claim 5,
Cultivation method for increasing the content of the mycelium of Cordyceps sinensis, characterized in that LED green light having a wavelength of 500nm to 550nm represents the maximum growth amount of the cordyceps mycelium. The method of claim 4,
A culture method for increasing the content of mycelium Cordyceps sinensis, characterized in that the mycelium is cultured by irradiating the inoculated medium with LED single light or mixed light for 5 to 15 hours per day. The method of claim 1,
The dry weight of the mycelium of Cordyceps sinensis,
A cultivation method for increasing the content of mycelium Cordyceps sinensis, characterized in that it has a correlation as shown in Equation 1 below with the amount of light, the glucose content of the medium, and the culture time:
[Equation 1]
Y ₁ =14.53-1.14X ₁ -0.9713X ₂ +0.5025X ₃ -0.9925X ₁ X ₂ +0.5450X ₁ X ₃ +0.0100X ₂ X ₃ +2.65X ₁ ² +2.19X ₂ ² +2.58X ₃ ²
(In Equation 1, Y ₁ is the dry weight (g/L) of the mycelium of Cordyceps sinensis, X ₁ is the amount of light, X ₂ is the glucose content of the medium, and X ₃ is the culture time.) The method of claim 8,
Culture method for increasing the content of the mycelium of Snow Cordyceps sinensis, characterized in that the amount of light, the glucose content of the medium, and the culture conditions of the culture time are 1000 to 1500 lux, 1 to 10 g/50 mL, and 80 to 120 hours, respectively. The method of claim 9,
Culture method for increasing the content of the mycelium of Cordyceps sinensis, characterized in that the amount of light, the glucose content of the medium, and the culture conditions of the culture time are 1335.16 lux, 5.01181 g/50mL, and 97.6614 hours, respectively. The mycelium of Snow Cordyceps sinensis cultured according to the method of any one of claims 1 to 10. In the method for predicting the dry weight of the mycelium of Cordyceps sinensis according to claim 1,
(a) Experiment by coding as -1, 0 and 1 for the amount of light (X ₁ ), the glucose content of the medium (X ₂ ), and the incubation time (X _{3) by Box-Behnken design} Designing a range;
(b) obtaining experimental values for the amount of light, the glucose content of the medium, and the incubation time in the experimental range designed in step (a);
(c) deriving a quadratic regression model represented by Equation 1 below using the experimental value of step (b); And
(d) predicting the dry weight of the mycelium of Cordyceps sinensis by variate analysis (ANOVA) of the quadratic regression model represented by Equation 1 derived in step (c); Dry weight prediction method of mycelium Cordyceps sinensis, characterized in that it comprises:
[Equation 1]
Y ₁ =14.53-1.14X ₁ -0.9713X ₂ +0.5025X ₃ -0.9925X ₁ X ₂ +0.5450X ₁ X ₃ +0.0100X ₂ X ₃ +2.65X ₁ ² +2.19X ₂ ² +2.58X ₃ ²
(In Equation 1, Y ₁ is the dry weight (g/L) of the mycelium of Cordyceps sinensis, X ₁ is the amount of light (code unit), X ₂ is the glucose content of the medium (code unit), and X ₃ is the culture time (code unit). Means.) The method of claim 12,
The amount of light, the glucose content of the medium and the culture conditions of the culture time are 1000 to 1500 lux, 1 to 10 g/50 mL, and 80 to 120 hours, respectively. The method of claim 13,
The amount of light, the glucose content of the medium, and the culture conditions of the culture time are 1335.16 lux, 5.01181 g/50mL, and 97.6614 hours, respectively.

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