TWI753534B - Solid state fermentation method of hispidin - Google Patents

Abstract

The present disclosure provides a solid state fermentation method of hispidin including the following steps. A culture material is provided, wherein the culture material includes a cereal grain medium. A fungus activation step is performed, wherein a Phellinus linteusfungi is inoculated into a liquid medium for an activation time so as to obtain a Phellinus linteusfungi solution. An inoculation step is performed, wherein the Phellinus linteusfungi solution is inoculated into the culture material so as to obtain a mixture. A culture step is performed, wherein the mixture is cultured at a culture temperature for a culture time so as to obtain a fermentation culture, wherein the fermentation culture includes hispidin. Therefore, the solid state fermentation method of hispidin of the present disclosure can rapidly and effectively produce hispidin in mass production, so that the solid state fermentation method of hispidin of the present disclosure has application potential in the relevant market.

Description

Solid-state fermentation method of sclerostin

The present invention relates to a solid state fermentation method, in particular to a solid state fermentation method of sclerostin.

Hispidin (Hispidin, 2-[(E)-2-(3,4-dihydroxyphenyl)ethenyl]-6-hydroxypyran-4-one) is a secondary metabolite derived from fungi, which has anticancer, Antioxidant, antiviral, anti-dementia, anti-diabetic, anti-obesity and other biological activities are of great benefit to human health, which in turn makes sclerostin have relevant potential in the health care market.

The sclerotin is derived from the fruiting bodies, mycelia or fermentation broths of Pellinus sp. and Inonotus sp. strains. However, the content of sclerotin in the fruiting bodies, mycelia and fermentation broths of Xylinophora and Pseudomonas is not high, and the growth rate of the fruiting bodies is very slow. It takes 4 to 10 months, which leads to the fact that the preparation of sclerotin by using Syringomyces or Phosporus as a raw material not only requires a lot of manpower, material resources and time costs, but also cannot be mass-produced.

Therefore, how to develop a preparation method that can quickly and increase the yield of bristle for relevant applications is a technical subject with economic value.

One embodiment of the present invention is to provide a solid-state fermentation method for hispidin, comprising the following steps. A culture material is provided, wherein the aforementioned culture material comprises a grain medium and a metal ion additive, and a water content of the culture material is 50%-70% by weight. Carry out a bacterial classification activation step, and it is to inoculate a Phellinus linteus in a liquid medium and cultivate an activation time, so as to obtain a Phellinus linteus bacterial liquid, wherein the aforementioned activation time is 2 Week, and the accession number of the aforesaid Xylophora schizophrenia is BCRC 930211. Carry out an inoculation step, and it is inoculated in the aforementioned culturing raw material with the described Xylomicron bacterium liquid to obtain a mixture, wherein the inoculum amount of the described Xylomicron bacterium liquid in one of the culturing materials is 5 %~15% by weight. A culturing step is performed by culturing the aforementioned mixture at a culturing temperature for 6 to 8 weeks to obtain a fermented culture, and the aforementioned fermented culture contains a sclerostin.

According to the above-mentioned solid-state fermentation method of sclerostin, the above-mentioned grain culture medium may comprise a brown rice, a rye or a combination thereof.

According to the aforementioned solid state fermentation method of bristle, the aforementioned metal ion additive may comprise KH ₂ PO ₄ , ZnCO ₃ or a combination thereof.

According to the aforementioned solid-state fermentation method of sclerostin, a mass percentage based on the culture material is 100%, and a mass percentage of the aforementioned metal ion additive can be 0.01% to 0.1%.

According to the above-mentioned solid state fermentation method of bristle, wherein the above-mentioned culture temperature can be 20 ℃ to 25 ℃.

Another embodiment of the present invention is to provide a method for solid-state fermentation of sclerostin, comprising the following steps. A culture material is provided, wherein the aforementioned culture material comprises a grain culture medium, and a water content of the culture material is 50%-70% by weight. Carry out a bacterial classification activation step, and it is to inoculate a Xylomicronia schizophrenia in a liquid medium and cultivate an activation time to obtain a Xylomicronia bacterium liquid, wherein the aforementioned activation time is 2 weeks, and the aforementioned The accession number of Xyloporus cleft hoof is BCRC 930211. Carry out an inoculation step, and it is inoculated in the aforementioned culturing raw material with the described Xylomicron bacterium liquid to obtain a mixture, wherein the inoculum amount of the described Xylomicron bacterium liquid in one of the culturing materials is 5 %~15% by weight. A culturing step is performed by culturing the aforementioned mixture at a culturing temperature for 6 to 8 weeks to obtain a fermented culture, and the aforementioned fermented culture contains a sclerostin. Wherein, the aforementioned culturing step includes a first temperature difference culturing step and a second temperature difference culturing step. The culturing temperature of the first temperature difference culturing step is 20°C to 25°C. The culturing temperature of the second temperature difference culturing step is 16°C to 19°C. Wherein, the aforesaid mixture is cultivated in the first temperature difference culturing step until one of the mycelium of Xylomicron is completely entangled with the aforesaid mixture.

According to the above-mentioned solid-state fermentation method of sclerostin, the above-mentioned grain culture medium may comprise a brown rice, a rye or a combination thereof.

Another embodiment of the present invention is to provide a method for solid-state fermentation of sclerostin, comprising the following steps. A culture material is provided, wherein the aforementioned culture material comprises a grain culture medium, and a water content of the culture material is 50%-70% by weight. Carry out a bacterial classification activation step, and it is to inoculate a Xylomicronia schizophrenia in a liquid medium and cultivate an activation time to obtain a Xylomicronia bacterium liquid, wherein the aforementioned activation time is 2 weeks, and the aforementioned The accession number of Xyloporus cleft hoof is BCRC 930211. Carry out an inoculation step, and it is inoculated in the aforementioned culturing raw material with the described Xylomicron bacterium liquid to obtain a mixture, wherein the inoculum amount of the described Xylomicron bacterium liquid in one of the culturing materials is 5 %~15% by weight. A culturing step is performed by culturing the aforementioned mixture at a culturing temperature for 6 to 8 weeks to obtain a fermented culture, and the aforementioned fermented culture contains a sclerostin. Wherein, the aforementioned culturing temperature is 20°C to 25°C, and the aforementioned culturing step comprises a first illumination culturing step and a second illumination culturing step. The first step of culturing in light is to cultivate the aforementioned mixture in the dark or under continuous illumination. The second light incubation step was performed under a 12-hour light-12-hour dark photoperiod or continuous light. Wherein, the aforesaid mixture is cultivated in the first light culturing step until one of the mycelia of Xyloporus schizophrenia is completely entangled with the aforesaid mixture.

According to the aforementioned solid-state fermentation method for sclerostin, one of the aforementioned second light culturing step may be 450 lux to 550 lux.

According to the above-mentioned solid-state fermentation method of sclerostin, the above-mentioned grain culture medium may comprise a brown rice, a rye or a combination thereof.

Yet another embodiment of the present invention is to provide a method for solid-state fermentation of sclerostin, comprising the following steps. A culture material is provided, wherein the culture material comprises a grain, and an initial pH value of one of the culture materials is 5.5 to 6.5. Carry out a bacterial classification activation step, and it is to inoculate a Xylem in a liquid medium and cultivate for an activation time, to obtain a Xylomicron bacterium liquid, wherein the deposit number of the aforementioned Xylomycium for BCRC 930211. Carry out an inoculation step, and it is inoculated with the aforementioned Xylomicron bacterium liquid in the aforementioned culturing material, to obtain a mixture, wherein the aforementioned Xylomicron bacterium liquid is 5%～5% in one of the culture materials. 15% by weight. Carry out a culturing step, it is a cultivation time with the aforementioned mixture at a culturing temperature, dark-cultivated, to obtain a fermentation culture, wherein the aforementioned culturing temperature is 20 ℃ to 25 ℃, and the aforementioned cultivating time is 6 Week to 8 weeks, and the monosclerotin content of the aforementioned fermentation culture is 0.055 mg/g to 1.110 mg/g.

According to the aforementioned solid-state fermentation method of bristle, the initial pH value of the aforementioned culture material can be 5.5.

According to the above-mentioned solid-state fermentation method of sclerostin, the water content of one of the above-mentioned culture materials may be 50% to 60% by weight. Preferably, the water content of the aforementioned culture material may be 55% by weight.

According to the aforementioned solid state fermentation method of sclerostin, the aforementioned grains can be selected from the group consisting of brown rice, millet, black rice, rye, oat, buckwheat, wheat and barley.

According to the above-mentioned solid-state fermentation method of sclerostin, the inoculation amount of the above-mentioned Xyloporus schizophrenia bacterial solution in the culture material can be 10% by weight.

According to the aforementioned solid-state fermentation method of bristle, the aforementioned activation time may be 1 week.

According to the aforementioned solid state fermentation method of sclerostin, the aforementioned liquid medium can be PDB medium (Potato dextrose broth), MEB medium (Malt extract broth) or YGB medium (Yeast extract glucose broth).

According to the aforementioned solid state fermentation method of bristle, the aforementioned culture material may further comprise a carbon source, and the aforementioned carbon source may be glucose, mannose or sucrose.

According to the above-mentioned solid-state fermentation method of sclerostin, the above-mentioned culture material can further comprise a nitrogen source, and the above-mentioned nitrogen source can be yeast extract, malt extract or protein gluten.

Thereby, the solid-state fermentation method of sclerostin of the present invention carries out solid-state fermentation with novel Xyloporus cleft hoof, and by adding metal ion additives to the culture raw materials, changing the temperature difference conditions of solid-state fermentation, and changing the light conditions of solid-state fermentation Or match the initial pH value, inoculum amount and other culturing steps of suitable culture materials, which will effectively obtain sclerostin in the fermented culture in a short time and improve the sclerostin content in the fermented culture, and then make the sclerostin in the fermentation culture. The solid-state fermentation method for pilosin of the present invention can rapidly and efficiently increase the yield of bristle, and has the development potential of the relevant market.

The following will illustrate specific test examples of the present invention with reference to the drawings, so that those skilled in the art to which the present invention pertains can fully utilize and practice the present invention without excessive interpretation and experimentation. However, the reader should understand that these practical details should not be used to limit the present invention, that is, in some experimental examples of the present invention, these practical details are not necessary, but are used to illustrate how to implement the present invention materials and methods.

One, the solid-state fermentation method of sclerostin of the present invention

Please refer to FIG. 1 , which is a flow chart showing the steps of a solid-state fermentation method 100 for sclerostin according to an example of an embodiment of the present invention. The solid-state fermentation method 100 of sclerostin includes step 110 , step 120 , step 130 and step 140 .

Step 110 is to provide a culture material, wherein the culture material includes a grain medium and a metal ion additive, and a water content of the culture material is 50%-70% by weight. In detail, the grain medium of the culture material of the present invention may comprise brown rice, rye or a combination thereof, and the metal ion supplement may comprise KH ₂ PO ₄ , ZnCO ₃ or a combination thereof.

When preparing the culture material, the metal ion additive will be first dissolved in water and mixed with the grain medium in an appropriate ratio, and the water content will be further adjusted to be 50% to 70% by weight, then at 121 ° C, 1.5 Kg/cm ² After sterilization under the conditions of 30 minutes, it is cooled to room temperature to obtain the culture material of the present invention, which is required for subsequent preparation. Furthermore, a mass percentage of the culture material based on the present invention is 100%, and a mass percentage of the metal ion additive may be 0.01% to 0.1%. In addition, a mass percentage of the culture material based on the present invention is 100%, and a mass percentage of the metal ion additive can be 0.05%.

Step 120 is to carry out a strain activation step, which is to inoculate a Phillinus linteus in a liquid medium and cultivate for an activation time, so as to obtain a Phillinus linteus bacterial solution, wherein the aforementioned Phillinus linteus The accession number of Xylophylla is BCRC 930211. In detail, the aforesaid Xylomicronia was collected and isolated by the inventor, and it was deposited in the Food Industry Development Research Institute (Taiwan, Hsinchu), and the deposit number is BCRC 930211, And the following description will be made by simply referring to the "Xyloporus schizophrenia of the present invention" whose deposit number is BCRC 930211.

In the aspect of the bacterial strain activation step, firstly, the mycelial block with a size of 3 cm × 3 cm was cut from the P. schizophrenia of the present invention stored at 4 ° C, and the aforementioned mycelial block was inoculated into a After the Erlenmeyer flask of the liquid culture medium, under the condition of 120 rpm, 25 DEG C, the shaking culture is carried out for 7 days to obtain the Xyloporus schizophrenia bacterial liquid of the present invention. The aforementioned liquid medium can be PDB medium (Potato dextrose broth), MEB medium (Malt extract broth) or YGB medium (Yeast extract glucose broth). Furthermore, the activation time of the present invention for culturing Xylinomycetes in the aforementioned liquid medium can be 2 weeks, so as to activate the physiological and biochemical activities of the present invention Xylinophora.

Step 130 is to carry out an inoculation step, which is to inoculate the above-mentioned culturing raw material with the strain of Xylomicron of the present invention, so as to obtain a mixture, wherein the strain of Xylomicron of the present invention is inoculated in one of the culture materials The amount is 5% to 15% by weight.

Step 140 is a culturing step of culturing the aforementioned mixture at a culturing temperature for 6 to 8 weeks to obtain a fermented culture, and the fermented culture contains sclerostin. Wherein, the aforementioned culture temperature may be 20°C to 25°C.

Thereby, the solid-state fermentation method 100 of sclerostin of the present invention includes grain culture medium and metal ion additives in the culture material, and performs solid-state fermentation with the Schizophrenia schizophrenia of the present invention, which can effectively improve the fermentation culture. The sclerostin content, so that the sclerostin solid-state fermentation method 100 of the present invention can quickly and efficiently generate sclerostin, and has the development potential of the relevant market.

Please refer to FIG. 2 , which is a flow chart showing the steps of a solid-state fermentation method 200 for sclerostin according to another embodiment of the present invention. The solid-state fermentation method 200 of sclerostin includes steps 210, 220, 230, and 240, wherein steps 210, 220, and 230 are similar to steps 110, 120, and 130 in FIG. The culturing material of the solid-state fermentation method 200 of Maosu does not contain metal ion additives, and the culturing temperature is also different, so the other same culturing details will not be repeated here.

The step 240 includes a first temperature difference culturing step 241 and a second temperature difference culture step 242, wherein the culture temperature of the first temperature difference culture step 241 is 20°C to 25°C, and the culture temperature of the second temperature difference culture step 242 is 16°C. °C to 19°C. In detail, the solid-state fermentation method 200 of the sclerostin of the present invention is to carry out solid-state fermentation on the Xylomicron of the present invention under different temperature difference conditions, and the mixture comprising the Xylomicron of the present invention will first be heated at 20° C. Cultivate under the conditions of C to 25 ° C until the hyphae of Xyloporus schizophrenia of the present invention are completely entangled on the mixture, and then move the completely entangled mixture with hyphae to an environment of 16 ° C to 19 ° C The culture is continued to further enhance the ability of Xyloporus schizophrenia of the present invention to entangle on the culture material, and can effectively generate sclerotin in the fermented culture.

Thereby, the solid-state fermentation method 200 of sclerostin of the present invention can effectively generate sclerostin in the fermented culture by changing the temperature difference condition of solid-state fermentation, and carry out solid-state fermentation with the Schizophrenia sp. By increasing the sclerostin content of the fermentation culture, the solid-state fermentation method 200 for sclerostin of the present invention can quickly and efficiently generate sclerostin, and has the development potential of the relevant market.

Please refer to FIG. 3 again, which is a flow chart showing the steps of a method 300 for solid-state fermentation of sclerostin according to another embodiment of the present invention. The solid-state fermentation method 300 of sclerostin includes steps 310, 320, 330, and 340, wherein steps 310, 320, and 330 are similar to steps 110, 120, and 130 in FIG. The culturing material of the solid-state fermentation method 300 of Maosu does not contain metal ion additives, so other same culturing details will not be repeated here.

The step 340 includes a first illumination cultivation step 341 and a second illumination cultivation step 342, wherein the first illumination cultivation step 341 is to cultivate the aforementioned mixture in the dark or under continuous illumination, and the mixture is cultivated under the first illumination. Step 341 is culturing until the mycelium of Xyloporus schizophrenia of the present invention is completely entangled on the mixture. Next, the mixture completely entangled with mycelium is cultured in the second light culture step 342 under a photoperiod of 12 hours light and 12 hours dark light or continuous light, so as to stimulate the Clefthoof of the present invention with different light conditions The ability of Laminaria to metabolize sclerostin. Wherein, one of the light illuminance in the second light cultivation step 342 may be 450 lux to 550 lux, but the present invention is not limited to this.

Thereby, the solid-state fermentation method 300 of the sclerostin of the present invention can effectively generate sclerostin in the fermentation culture by changing the light conditions of the solid-state fermentation, and performing the solid-state fermentation with the Schizophrenia sp. of the present invention, Thus, the solid-state fermentation method 300 of the present invention for sclerostin can quickly and efficiently generate sclerostin, and has the development potential of the relevant market.

Please refer to FIG. 4 , which is a flow chart showing the steps of a solid-state fermentation method 400 of sclerostin according to still another embodiment of the present invention. The solid state fermentation method 400 of sclerostin includes step 410 , step 420 , step 430 and step 440 .

Step 410 is to provide a culture material, wherein the culture material includes a grain, and an initial pH value of one of the culture materials is 5.5 to 6.5. In detail, the culture material includes grains and water, and when preparing the culture material, the pH value of the water will be adjusted to 5.5 to 6.5, and then thoroughly mixed with the grains, and then heated at 121°C and 1.5 Kg/cm ² . Condition sterilization for 30 minutes and then cooled to room temperature to obtain a culture material with an initial pH value of 5.5 to 6.5. Preferably, the initial pH value of the culture material can be 5.5, and the grain can be brown rice, millet, black rice, rye, oat, buckwheat, wheat or barley. In terms of the water content of the culture material, the water content of the culture material can be 50% to 60% by weight, preferably 55% by weight, to provide for subsequent culture.

Step 420 is to carry out a bacterial classification activation step, and it is to inoculate a Xylem in a liquid medium and cultivate for an activation time, so as to obtain a bacteria liquid of X. It is the Schizophrenia of the present invention.

In the aspect of the bacterial strain activation step, firstly, the mycelial block with a size of 2 cm × 2 cm was cut from the S. pyogenes of the present invention stored at 4°C, and the aforementioned mycelial block was inoculated into a 100 mL of After the Erlenmeyer flask of the liquid culture medium, under the condition of 120 rpm, 25 DEG C, the shaking culture is carried out for 7 days, so as to obtain the Xylomicron bacterium liquid of the present invention. Preferably, the aforementioned liquid medium can be PDB medium, MEB medium or YGB medium. Furthermore, the Xylomya cleft hoof of the present invention will be cultured in the aforementioned liquid medium for 1 to 2 weeks to activate the physiological and biochemical activities of the Xylomya cleft hoof of the present invention.

Step 430 is to carry out an inoculation step, which is to inoculate the above-mentioned culturing raw material with the Xylomicron of the present invention to obtain a mixture, wherein the Xylomicron of the present invention is inoculated in one of the culture materials The amount is 5% to 15% by weight. Preferably, the inoculation amount of the Xyloporus schizophrenia bacterial solution of the present invention in the culture material can be 10% by weight.

Step 440 is to carry out a culturing step, which is to cultivate the aforementioned mixture in the dark for a culturing time at a culturing temperature to obtain a fermented culture, wherein the aforementioned culturing temperature is 20°C to 25°C, and the aforementioned culturing The time period was 6 to 8 weeks, and the monosclerotin content of the fermentation culture was 0.055 mg/g to 1.110 mg/g.

In addition, the culture material of the solid-state fermentation method 400 of sclerostin of the present invention may further comprise a carbon source or a nitrogen source, wherein the carbon source may be glucose, mannose or sucrose, and the nitrogen source may be yeast extract, malt The extract or the protein extract can improve the growth state and activity of Xyloporus schizophrenia of the present invention, thereby increasing the production efficiency of the solid-state fermentation method 400 of the sclerostin of the present invention.

After the cultivation in step 410, step 420, step 430 and step 440, the Xyloporus cleft hoof of the present invention can entangle and proliferate on the culture material, and can effectively generate and improve the sclerostin in the fermented culture. Therefore, the solid-state fermentation method 400 of the present invention can rapidly and efficiently mass-produce sclerostin, so as to provide relevant application requirements, and has the development potential of health care and medical related markets.

2. Examples and Comparative Examples

1. Effects of metal ion supplements and culture conditions on sclerotin content

Specific examples of the present invention will be presented below to describe in detail the sclerostin content in the fermented culture of the solid-state fermentation method of sclerostin of the present invention under different grain culture media, different metal ion supplements and different culture conditions , and the culture conditions of each example and the control group in this experiment are listed in Table 1. Wherein, each embodiment, each comparative example and each control group of this experiment are all cultivated by the solid-state fermentation method of sclerostin of the present invention, so please refer to the previous paragraph for the relevant cultivation details, which will not be repeated here. . Table I culture material Schizophrenia of the present invention Cultivation steps grain culture medium metal ion additives Moisture content (%) Inoculum amount (%) liquid medium Incubation temperature (°C) Lighting conditions Example 1 brown rice 0.05% KH ₂ PO ₄ 55 10 PDB 25 Avoid light Example 2 brown rice 0.05% ZnCO ₃ 55 10 PDB 25 Avoid light Example 3 rye 0.05% KH ₂ PO ₄ 55 10 PDB 25 Avoid light Example 4 rye 0.05% ZnCO ₃ 55 10 PDB 25 Avoid light Example 5 brown rice - 55 10 PDB 25 Avoid light 18 Example 6 brown rice - 55 10 PDB 25 Avoid light 18 (5 days) 25 Example 7 rye - 55 10 PDB 25 Avoid light 18 Example 8 rye - 55 10 PDB 25 Avoid light 18 (5 days) 25 Example 9 brown rice - 55 10 PDB 25 Avoid light 12 hours light 12 hours dark Example 10 brown rice - 55 10 PDB 25 continuous light Example 11 rye - 55 10 PDB 25 Avoid light 12 hours light 12 hours dark Example 12 rye - 55 10 PDB 25 continuous light control group 1 brown rice - 55 10 PDB 25 Avoid light control group 2 rye - 55 10 PDB 25 Avoid light

As shown in Table 1, Example 5 and Example 7 were cultured at 25°C before the hyphae of Xylomicronia were completely entangled with the mixture, and then moved to 18°C after the hyphae were completely entangled with the mixture. C, while Example 6 and Example 8 were cultured at 25°C before the hyphae of Xylomicron was completely entangled with the mixture, and then moved to 18°C after the hyphae were completely entangled with the mixture. Cultivated at °C for 5 days, and then further transferred to a 25 °C environment to continue the culture. In addition, in Example 9 and Example 11, before the hyphae of Xylomicron is completely entangled with the mixture, the culture was carried out in the dark, and after the hyphae were completely entangled with the mixture, the culturing was carried out under a light cycle of 12 hours of light and 12 hours of darkness. nourish.

For the determination of mycelial content, indirect analysis was performed with the ergosterol concentration of the fermented culture powder. In the experiment, 1 g of the fermentation culture powders of Example 1 to Example 12 were placed in different glass test tubes, and 4 mL of n-hexane was added. After shaking and extracting for 90 seconds, centrifuge and collect the supernatant. After repeated extraction to remove the supernatant precipitate twice, the supernatant collected from the three extractions was concentrated under reduced pressure at 40°C, and methanol was added to the sample after the concentration under reduced pressure was quantified to 5. mL to obtain the experimental samples corresponding to the fermentation culture powders of Examples 1 to 12. The aforementioned experimental samples will be filtered with a 0.45 μm membrane and then analyzed by high performance liquid chromatography (HPLC). The detection conditions of the high performance liquid chromatography method are listed in Table 2. The above experiment will be repeated three times, and the results of the three experiments will be averaged to evaluate the content of ergosterol in Example 1 to Example 12, and the content of mycelium is further determined by the difference between ergosterol and mycelium. By calculating the calibration curve, the mycelium content values of Examples 1 to 12 can be obtained. Table II high performance liquid chromatography pump SHIMADZU LC-20AT Detector SHIMADZU SPD-20A UV-VIS DETECTOR (Visible/Ultraviolet) Separation column Kinetex 5µC18 100A (250 × 4.6 mm, 5 μm, Phenomenex) Injection volume (μL) 20 mobile phase 100% methanol Detection wavelength (nm) 282 (UV) Flow rate (mL/min) 1.2

In the determination of the sclerostin content of the fermentation culture, firstly, 0.2 g of the fermentation culture powders of Examples 1 to 12 were placed in glass test tubes, and then 10 mL of methanol was added to extract with ultrasonic vibration for 1 hour, and then After 1 hour of shaking extraction, the extract was filtered with a 0.45 μm filter membrane, and the content of sclerotin was analyzed by high performance liquid chromatography. Please refer to Table 2 for the detection conditions of high performance liquid chromatography. The above experiments will be performed in triplicate, and the results of the three experiments will be averaged to evaluate the sclerotin content of the fermentation cultures of Examples 1 to 12. And the specific productivity of sclerotin of the mycelium of embodiment 1 to embodiment 12 can be further based on the aforementioned mycelium content analysis value and sclerotin content analysis value and utilize the calculation formula 1 of sclerotin specific productivity Calculated, the calculation formula 1 of bristle specific productivity is as follows: Sclerotin specific yield (%) = ( sclerostin content of fermented cultures ) × 100 % (Formula I). Mycelium content of fermentation cultures

In addition, the present invention further comprises Comparative Example 1 to Comparative Example 8, which are culturing Xyloporus schizophrenia of the present invention with the culture material of adding different metal ion additives, to further illustrate the solid-state fermentation method of sclerostin of the present invention The sclerotin content of fermentation cultures after addition of various metal ion supplements. Wherein, comparative example 1 to comparative example 8 all carry out solid-state culture with the culturing material of water content 55% by weight, and the present invention is cultivated with PDB liquid medium, wherein The inoculum amount of the bacterial liquid in one of the culturing raw materials is 10% by weight, and the whole process is cultivated in the dark under the condition of 25 ° C, and the types and additions of the metal ion additives of Comparative Example 1 to Comparative Example 8 are listed in Table III. Table 3 culture material grain culture medium metal ion additives Metal ion additive content (%) Comparative Example 1 brown rice Na ₂ CO ₃ 0.05 Comparative Example 2 brown rice MgSO ₄ ˙7H ₂ O 0.05 Comparative Example 3 brown rice FeSO ₄ ˙7H ₂ O 0.05 Comparative Example 4 brown rice _CaCO3 0.05 Comparative Example 5 rye Na ₂ CO ₃ 0.05 Comparative Example 6 rye MgSO ₄ ˙7H ₂ O 0.05 Comparative Example 7 rye FeSO ₄ ˙7H ₂ O 0.05 Comparative Example 8 rye _CaCO3 0.05

＜Effect of metal ion additives on sclerotin production＞

In this experiment, the aforementioned control group 1, control group 2, examples 1 to 4, and comparative examples 1 to 8 are used to conduct experiments to analyze whether the addition of metal ion additives and the types of additions affect the bristles of the present invention. The effect of sclerostin yields on solid-state fermentation of gluten.

Please refer to Table 4, Table 4 is the mycelium content, sclerotin content and sclerotin specific yield of the present invention's Xyloporus spp. grown under the brown rice medium of adding different metal ion additives. Table 4 Cultivation time (weeks) 4 5 6 7 8 control group 1 Mycelium content (mg/g) 81.11±8.64 149.98±12.22 187.35±12.93 197.57±17.63 198.52±16.75 Hardhair content (mg/g) 0.036±0.004 0.110±0.004 0.340±0.014 0.165±0.003 0.045±0.004 The specific yield of sclerotin (%) 0.044±0.002 0.073±0.003 0.181±0.008 0.084±0.003 0.023±0.007 Example 1 Mycelium content (mg/g) 80.15±6.52 150.58±11.17 185.27±18.96 202.55±14.20 199.38±15.35 Hardhair content (mg/g) 0.000±0.000 0.003±0.000 0.068±0.004 0.059±0.001 0.025±0.001 The specific yield of sclerotin (%) 0.000±0.000 0.002±0.000 0.037±0.006 0.029±0.001 0.013±0.001 Example 2 Mycelium content (mg/g) 76.36±2.18 150.25±9.15 180.25±17.35 195.90±12.27 196.96±15.35 Hardhair content (mg/g) 0.236±0.005 0.582±0.046 0.306±0.016 0.094±0.041 0.080±0.006 The specific yield of sclerotin (%) 0.309±0.001 0.387±0.007 0.170±0.005 0.048±0.022 0.041±0.006 Comparative Example 1 Mycelium content (mg/g) 1.30±0.28 2.60±0.15 3.51±0.62 3.45±1.20 3.62±0.25 Hardhair content (mg/g) 0.000±0.000 0.000±0.000 0.000±0.000 0.000±0.000 0.000±0.000 The specific yield of sclerotin (%) 0.000±0.000 0.000±0.000 0.000±0.000 0.000±0.000 0.000±0.000 Comparative Example 2 Mycelium content (mg/g) 74.36±5.16 132.39±11.28 176.68±12.20 185.57±18.87 190.15±16.30 Hardhair content (mg/g) 0.001±0.000 0.001±0.000 0.000±0.000 0.000±0.000 0.000±0.000 The specific yield of sclerotin (%) 0.001±0.000 0.001±0.000 0.000±0.000 0.000±0.000 0.000±0.000 Comparative Example 3 Mycelium content (mg/g) 71.21±6.67 130.57±9.42 169.35±13.63 179.77±17.30 188.65±17.72 Hardhair content (mg/g) 0.000±0.000 0.001±0.000 0.002±0.000 0.000±0.000 0.000±0.000 The specific yield of sclerotin (%) 0.000±0.000 0.001±0.000 0.001±0.000 0.000±0.000 0.000±0.000 Comparative Example 4 Mycelium content (mg/g) 78.38±3.77 151.57±9.38 186.16±16.20 199.95±19.30 198.08±27.14 Hardhair content (mg/g) 0.021±0.004 0.048±0.002 0.091±0.007 0.060±0.008 0.035±0.006 The specific yield of sclerotin (%) 0.027±0.009 0.032±0.003 0.048±0.007 0.030±0.009 0.018±0.003

As can be seen from the content of Table 4, under the situation of taking brown rice culture medium as the grain culture medium of culturing raw material, the sclerotin yield of the control group 1 that does not add metal ion additive is the highest in the 6th week, and its sclerostin unit content is 0.340. mg/g, the specific yield of sclerostin can reach 0.181%. In terms of adding different metal ion additives, the sclerotin yield of Example 2 was the highest, the sclerotin unit content of which was cultured to the 5th week was 0.582 mg/g, and the sclerosin specific yield could reach 0.387% , it shows that in the case of using brown rice medium as the grain medium for culturing raw materials, selecting KH ₂ PO ₄ as the metal ion additive of the present invention can effectively increase the sclerostin content in the fermented culture.

Please refer to Table 5, Table 5 is the mycelium content, sclerotin content and sclerotin specific yield of the present invention's Xyloporus schizophrenia growing under the rye culture medium of adding different metal ion additives. Table 5 Cultivation time (weeks) 4 5 6 7 8 control group 2 Mycelium content (mg/g) 73.97±7.85 154.47±13.73 178.48±11.82 195.19±16.92 199.20±20.02 Hardhair content (mg/g) 0.051±0.005 0.112±0.016 0.302±0.020 0.161±0.023 0.065±0.009 The specific yield of sclerotin (%) 0.069±0.009 0.073±0.012 0.169±0.005 0.082±0.010 0.033±0.008 Example 3 Mycelium content (mg/g) 71.85±6.75 147.27±15.70 175.47±13.72 205.10±21.29 201.96±16.62 Hardhair content (mg/g) 0.086±0.001 0.147±0.001 0.539±0.085 0.180±0.015 0.130±0.015 The specific yield of sclerotin (%) 0.120±0.002 0.100±0.001 0.307±0.012 0.088±0.009 0.064±0.010 Example 4 Mycelium content (mg/g) 76.36±6.12 157.27±13.72 176.65±18.34 195.64±15.53 196.76±17.21 Hardhair content (mg/g) 0.068±0.006 0.160±0.015 0.364±0.028 0.105±0.005 0.080±0.004 The specific yield of sclerotin (%) 0.089±0.007 0.102±0.008 0.206±0.008 0.054±0.004 0.041±0.004 Comparative Example 5 Mycelium content (mg/g) 75.85±8.17 157.21±11.53 180.18±15.65 199.22±17.70 194.71±22.45 Hardhair content (mg/g) 0.010±0.001 0.030±0.000 0.049±0.002 0.020±0.002 0.008±0.001 The specific yield of sclerotin (%) 0.013±0.005 0.019±0.001 0.027±0.004 0.010±0.008 0.004±0.000 Comparative Example 6 Mycelium content (mg/g) 76.26±8.85 160.20±13.54 182.17±17.76 201.47±17.52 197.17±18.30 Hardhair content (mg/g) 0.040±0.012 0.125±0.010 0.215±0.012 0.080±0.007 0.065±0.010 The specific yield of sclerotin (%) 0.052±0.024 0.078±0.007 0.118±0.005 0.040±0.008 0.033±0.014 Comparative Example 7 Mycelium content (mg/g) 75.86±13.74 158.93±15.87 180.83±13.75 199.04±14.96 198.85±23.53 Hardhair content (mg/g) 0.000±0.000 0.000±0.000 0.000±0.000 0.000±0.000 0.000±0.000 The specific yield of sclerotin (%) 0.000±0.000 0.000±0.000 0.000±0.000 0.000±0.000 0.000±0.000 Comparative Example 8 Mycelium content (mg/g) 74.45±6.72 153.31±15.27 172.95±11.63 187.75±13.63 190.17±17.75 Hardhair content (mg/g) 0.001±0.000 0.002±0.001 0.001±0.001 0.000±0.000 0.000±0.000 The specific yield of sclerotin (%) 0.001±0.000 0.001±0.000 0.001±0.000 0.000±0.000 0.000±0.000

As can be seen from the contents of Table 5, in the case of using the rye culture medium as the grain culture medium of the culturing raw material, the sclerostin output of the control group 2 without the addition of the metal ion additive is the highest in the 6th week, and its sclerostin unit content is 0.302 mg/g, the specific yield of sclerotin could reach 0.169%. In terms of adding different metal ion additives, the sclerotin yield of Example 3 was the highest, the sclerotin unit content of which was cultured to the 6th week was 0.539 mg/g, and the sclerosin specific yield could reach 0.307% , followed by embodiment 4, the sclerotin unit content of its culture to the 6th week is 0.364 mg/g, and the sclerotin specific yield can reach 0.206%, showing that the rye culture medium is used as the grain medium of the culture material. In this case, selecting KH ₂ PO ₄ and ZnCO ₃ as the metal ion additives of the present invention can effectively increase the sclerotin content in the fermentation culture.

<Influence of culture temperature on sclerostin production>

This experiment is based on the above-mentioned Example 5 to Example 8 to analyze the influence of the change of culture temperature on the sclerostin yield of the solid-state fermentation method of sclerostin of the present invention.

Please refer to Fig. 5 and Table 6, Fig. 5 is the mycelial growth situation of the present invention's Xylomicron under different temperature difference conditions, cultivated in brown rice medium, and Table 6 is the present invention's Xylem in different temperature differences. Mycelium content, sclerostin content and sclerostin specific yield cultured in brown rice medium under the condition of temperature difference. Table 6 Cultivation time (weeks) 4 5 6 7 8 Example 5 Mycelium content (mg/g) 70.65±7.52 139.46±10.33 160.72±14.45 185.52±10.53 189.40±12.89 Hardhair content (mg/g) 0.036±0.000 0.105±0.006 0.245±0.017 0.090±0.005 0.048±0.004 The specific yield of sclerotin (%) 0.051±0.001 0.075±0.004 0.152±0.006 0.048±0.003 0.025±0.005 Example 6 Mycelium content (mg/g) 72.35±6.29 142.01±11.37 165.64±10.77 187.65±16.65 189.92±19.85 Hardhair content (mg/g) 0.036±0.006 0.095±0.005 0.175±0.007 0.075±0.005 0.032±0.005 The specific yield of sclerotin (%) 0.050±0.013 0.067±0.004 0.106±0.002 0.040±0.004 0.017±0.005

As shown in Figure 5 and Table 6, when the brown rice medium is used as the grain medium of the culture material, the mycelium growth conditions of Example 5 and Example 6 are all good, and the sclerotin when cultured to the 6th week The unit content is 0.245 mg/g in Example 5 and 0.175 mg/g in Example 6, respectively, which shows that under the condition of using brown rice medium as the grain medium of the culture material, different temperature difference conditions will be effective in the fermentation culture. Hardhair.

Please refer to Fig. 6 and Table 7, Fig. 6 is the mycelial growth situation of Xylomycium sp. of the present invention cultivated in the rye medium under different temperature difference conditions, and Table 7 is the Mycelia of the present invention. Mycelium content, sclerostin content and sclerostin specific yield cultured in rye medium under different temperature conditions. Table 7 Cultivation time (weeks) 4 5 6 7 8 Example 7 Mycelium content (mg/g) 70.05±5.77 145.58±15.21 172.98±14.64 187.64±14.87 191.67±16.37 Hardhair content (mg/g) 0.036±0.008 0.085±0.007 0.235±0.021 0.102±0.022 0.043±0.009 The specific yield of sclerotin (%) 0.051±0.015 0.058±0.009 0.136±0.007 0.054±0.016 0.022±0.006 Example 8 Mycelium content (mg/g) 72.65±9.20 151.77±17.85 178.18±15.55 191.15±14.33 195.52±17.73 Hardhair content (mg/g) 0.030±0.002 0.076±0.006 0.134±0.012 0.085±0.008 0.035±0.004 The specific yield of sclerotin (%) 0.041±0.006 0.050±0.009 0.075±0.008 0.044±0.007 0.018±0.010

In this experiment, Example 7 was cultured at 25°C before the hyphae of Xylomicronia were completely entangled with the mixture, and then moved to 18°C after the hyphae were completely entangled with the mixture, while Example 8 was cultured at 25° C. before the hyphae of Xylomicronia were completely entangled with the mixture, and then moved to 18° C. for 5 days after the hyphae were completely entangled with the mixture, and then further transferred. The culture was continued at 25°C. As shown in Figure 6 and Table 7, when the rye culture medium was used as the grain culture medium of the culture material, the mycelium growth conditions of Example 7 and Example 8 were all good, and the bristles when cultured to the sixth week The unit content of the element is 0.253 mg/g in Example 7 and 0.134 mg/g in Example 8, respectively, which shows that when the rye medium is used as the grain medium of the culture material, different temperature differences will be effective in the fermentation culture. sclerostin is generated in the

＜Effect of light time on sclerostin production＞

This experiment is based on the above-mentioned Example 9 to Example 12 to analyze the effect of light time on the sclerostin yield of the solid-state fermentation method of sclerostin of the present invention.

Please refer to Fig. 7 and Table 8, Fig. 7 is the mycelial growth situation of Xylomicronia cleft hoof of the present invention cultivated in brown rice medium under different light conditions, and Table 8 is the hyphae growth situation of S. Mycelium content, sclerotin content and specific yield of sclerotin cultured in brown rice medium under light conditions. Table 8 Cultivation time (weeks) 4 5 6 7 8 Example 9 Mycelium content (mg/g) 84.60±15.15 148.96±8.65 191.07±14.52 195.69±18.32 200.68±14.35 Hardhair content (mg/g) 0.027±0.002 0.050±0.004 0.177±0.010 0.084±0.007 0.048±0.005 The specific yield of sclerotin (%) 0.032±0.004 0.034±0.004 0.063±0.004 0.043±0.005 0.024±0.003 Example 10 Mycelium content (mg/g) 85.82±15.98 146.36±10.55 186.75±16.83 195.50±8.54 198.57±12.72 Hardhair content (mg/g) 0.016±0.002 0.035±0.005 0.109±0.016 0.047±0.007 0.020±0.003 The specific yield of sclerotin (%) 0.019±0.003 0.024±0.003 0.058±0.007 0.024±0.003 0.010±0.01

In this test, Example 9 was cultured in the dark before the hyphae of Xylobacter sclerotiorum were completely entangled with the mixture, and after the hyphae were completely entangled with the mixture, the light was illuminated for 12 hours under the condition of illuminance of 550 lux. The 12-hour dark photoperiod was cultivated, while Example 10 was cultivated in an environment with a light illuminance of 550 lux throughout the whole process. As shown in Figure 7 and Table 8, the mycelial growth conditions of Example 9 and Example 10 are all good, and the sclerotin unit content when cultured to the 6th week is respectively 0.177 mg/g of Example 9 and implementation. The 0.109 mg/g of Example 10 shows that when brown rice medium is used as the grain medium for culturing raw materials, different light conditions can also effectively generate sclerostin in the fermentation culture.

Please refer to Fig. 8 and Table 9, Fig. 8 is the mycelial growth situation of Xylem in rye cultured in rye medium under different lighting conditions, and Fig. 8 is Xylem of the present invention in Mycelium content, sclerostin content and sclerostin specific yield in rye medium cultured under different light conditions. Table 9 Cultivation time (weeks) 4 5 6 7 8 Example 11 Mycelium content (mg/g) 82.25±18.65 146.47±13.55 180.05±14.75 189.93±17.85 194.25±15.90 Hardhair content (mg/g) 0.024±0.003 0.040±0.005 0.105±0.014 0.075±0.005 0.040±0.004 The specific yield of sclerotin (%) 0.029±0.005 0.027±0.003 0.058±0.006 0.039±0.003 0.021±0.002 Example 12 Mycelium content (mg/g) 80.66±10.37 146.86±7.59 177.52±12.87 186.71±8.65 190.22±13.75 Hardhair content (mg/g) 0.014±0.001 0.025±0.006 0.084±0.007 0.055±0.006 0.036±0.004 The specific yield of sclerotin (%) 0.017±0.002 0.017±0.002 0.047±0.004 0.029±0.002 0.019±0.002

In this test, Example 11 was cultured in the dark before the hyphae of Xylomicronia were completely entangled with the mixture, and after the hyphae were completely entangled with the mixture, the light was illuminated for 12 hours under the condition of illuminance of 550 lux for 12 hours. The cells were cultured in a light cycle of 1 hour in the dark, while Example 12 was cultured in an environment with an illumination intensity of 550 lux for the whole time. As shown in Figure 8 and Table 9, the mycelium growth conditions of Example 11 and Example 12 were all good, and the sclerostin unit content when cultured to the 6th week was 0.105 mg/g of Example 11 and 0.105 mg/g of Example 11 respectively. The 0.084 mg/g of Example 12 shows that when the rye medium is used as the grain medium of the culture material, different light conditions can also effectively generate sclerostin in the fermentation culture.

2. The effect of initial pH value of culture material and culture conditions on sclerostin content

Specific examples of the present invention will be presented below to describe in detail the grain types of the culture material, the initial pH value of the culture material, the water content of the culture material, and the pores of the cleft hoof wood layer of the present invention in the solid state fermentation method of sclerostin of the present invention The activation time of the bacteria, the liquid culture medium used in the step of activating the strain of Xylomicron of the present invention, and the inoculum amount of the strain of Xylomicron of the present invention in the culture material, and further analyze the bristles of the present invention The sclerostin content of the fermented culture and the sclerostin specific yield of the mycelium under the aforementioned different conditions of the solid-state fermentation method of gluten, and the culture conditions of each example in this experiment are listed in Table 10. Table 10 culture material Schizophrenia of the present invention cereals starting pH Moisture content (%) carbon source nitrogen source Activation time (weeks) liquid medium Inoculum amount (%) Example 13 brown rice 5.5 55 - - 1 PDB 10 Example 14 brown rice 6.5 55 - - 1 PDB 10 Example 15 brown rice 5.5 50 - - 1 PDB 10 Example 16 brown rice 5.5 60 - - 1 PDB 10 Example 17 Millet 5.5 55 - - 1 PDB 10 Example 18 black rice 5.5 55 - - 1 PDB 10 Example 19 wheat 5.5 55 - - 1 PDB 10 Example 20 Buckwheat 5.5 55 - - 1 PDB 10 Example 21 rye 5.5 55 - - 1 PDB 10 Example 22 oat 5.5 55 - - 1 PDB 10 Example 23 barley 5.5 55 - - 1 PDB 10 Example 24 brown rice 5.5 55 glucose - 1 PDB 10 Example 25 brown rice 5.5 55 Mannose - 1 PDB 10 Example 26 brown rice 5.5 55 sucrose - 1 PDB 10 Example 27 brown rice 5.5 55 - yeast extract 1 PDB 10 Example 28 brown rice 5.5 55 - Malt Extract 1 PDB 10 Example 29 brown rice 5.5 55 - protein gluten 1 PDB 10 Example 30 brown rice 5.5 55 - - 2 PDB 10 Example 31 brown rice 5.5 55 - - 3 PDB 10 Example 32 brown rice 5.5 55 - - 1 PDB 5 Example 33 brown rice 5.5 55 - - 1 PDB 15 Example 34 brown rice 5.5 55 - - 1 YGB 10 Example 35 brown rice 5.5 55 - - 1 MEB 10

In terms of the determination of the sclerostin content of the fermentation culture and the specific yield of sclerostin of the mycelium of the fermentation culture, firstly, the fermentation obtained by the solid-state fermentation method of the sclerostin of Example 13 to Example 35 was cultured. The culture was dried in an oven at 60°C to a constant weight, then pulverized with a pulverizer and sieved to obtain a fermented culture powder, and the aforementioned fermented culture powder will be further used to measure its mycelium content and sclerotin content. And the aspect of mycelium content determination and the sclerostin content determination of fermented culture is to utilize high performance liquid chromatography and the calculation formula I of the aforementioned sclerostin specific yield to analyze and calculate, and its operation method is the same as the previous paragraph. The description is the same and will not be repeated here.

In addition, the present invention further comprises Comparative Example 9 to Comparative Example 12, which are experiments with Xyloporus schizophrenia of the present invention under different culture conditions to further illustrate the fermentation culture of the solid-state fermentation method of sclerostin of the present invention. The content of sclerotin and the specific yield of sclerotin of mycelium, and the culture conditions of Comparative Examples 9 to 12 are listed in Table XI. Table 11 culture material Schizophrenia of the present invention cereals starting pH Moisture content (%) carbon source nitrogen source Activation time (weeks) liquid medium Inoculum amount (%) Comparative Example 9 brown rice 7.5 55 - - 1 PDB 10 Comparative Example 10 brown rice 5.5 55 fructose - 1 PDB 10 Comparative Example 11 brown rice 5.5 55 lactose - 1 PDB 10 Comparative Example 12 brown rice 5.5 55 - glutamic acid 1 PDB 10

<Analysis of the effect of the initial pH value of the culture material on the yield of sclerostin>

In this experiment, the above-mentioned Example 13, Example 14 and Comparative Example 9 were used to analyze the bristles of the fermentation culture obtained by the solid-state fermentation method of sclerostin of the present invention at different initial pH values of the culture materials. The ratio of sclerostin content to mycelium, and the relevant steps and details of the solid-state fermentation method of sclerostin of the present invention, please refer to the above, and will not be repeated here.

Please refer to Table 12, which serially shows the mycelium content, the sclerotin content and the sclerotin ratio of the mycelium of the fermented cultures of Example 13, Example 14 and Comparative Example 9 cultured for 5 to 8 weeks. Rate. As can be seen from the results in Table 12, the sclerotin content of Example 13 was the highest when the culture time was 6 weeks, and its sclerotin content was 0.350 mg/g, and the sclerosin specific yield of its mycelium reached 0.212 %, the sclerotin content of embodiment 14 was respectively 0.228 mg/g and 0.216 mg/g when the culture time was 7 weeks and 8 weeks, and the sclerotin specific productivity of its mycelium was 7 weeks respectively. 0.125% and 0.124% in the 8th week, and the sclerotin content of Example 13 and Example 14 compared with Comparative Example 9 and the sclerotin specific yield of mycelium are all good, showing the sclerotin of the present invention The solid-state fermentation method can effectively generate and increase the content of sclerostin in the fermentation culture under the condition that the initial pH value of the culture material is 5.5 to 6.5, so that it has the development potential of health care and medical related markets. Table 12 Cultivation time (weeks) 5 6 7 8 Example 13 Mycelium content (mg/g) 145.20±15.10 165.42±15.12 175.50±18.25 200.32±18.12 Hardhair content (mg/g) 0.093±0.006 0.350±0.014 0.227±0.014 0.104±0.004 The specific yield of sclerotin (%) 0.064±0.004 0.212±0.009 0.129±0.012 0.052±0.002 Example 14 Mycelium content (mg/g) 92.46±1.42 171.64±2.70 183.46±2.01 174.89±7.15 Hardhair content (mg/g) 0.082±0.012 0.128±0.012 0.228±0.001 0.216±0.003 The specific yield of sclerotin (%) 0.089±0.007 0.075±0.004 0.125±0.004 0.124±0.002 Comparative Example 9 Mycelium content (mg/g) 83.33±1.39 156.14±1.58 164.04±1.94 149.68±11.50 Hardhair content (mg/g) 0.056±0.005 0.075±0.004 0.088±0.002 0.069±0.002 The specific yield of sclerotin (%) 0.067±0.004 0.048±0.001 0.054±0.001 0.046±0.004

<Analysis of the influence of the water content of the culture material on the yield of sclerostin>

In this experiment, the above-mentioned Example 13, Example 15 and Example 16 are used to carry out the experiment to analyze the sclerotin content of the fermentation culture obtained by the solid-state fermentation method of sclerostin of the present invention when the water content of the culture material is different And the specific yield of sclerostin of mycelium, and the relevant steps and details of the solid-state fermentation method of sclerostin of the present invention refer to the above, and will not be repeated here.

Please refer to Table 13, which shows the mycelium content, the sclerotin content and the sclerotin ratio of the mycelium of the fermented cultures of Example 13, Example 15 and Example 16 in 5 to 8 weeks of culture. Rate. As can be seen from the results of Table 13, the sclerotin content of Example 13, Example 15 and Example 16 is the highest when the culture time is 6 weeks, and its sclerotin content is all higher than 0.270 mg/g, and the embodiment 13. The specific yields of sclerostin of the mycelia of Example 15 and Example 16 all reach more than 0.140%, indicating that the solid-state fermentation method of sclerostin of the present invention has a water content of 50% to 60% in the culture material. Under the condition of weight ratio, it can effectively generate and increase the content of sclerostin in the fermentation culture, so that it has the development potential of health care and medical related markets. Table 13 Cultivation time (weeks) 5 6 7 8 Example 13 Mycelium content (mg/g) 145.20±15.10 165.42±15.12 175.50±18.25 200.32±18.12 Hardhair content (mg/g) 0.093±0.006 0.350±0.014 0.227±0.014 0.104±0.004 The specific yield of sclerotin (%) 0.064±0.004 0.212±0.009 0.129±0.012 0.052±0.002 Example 15 Mycelium content (mg/g) 121.86±6.06 180.57±10.91 172.78±5.00 168.93±4.34 Hardhair content (mg/g) 0.081±0.005 0.252±0.040 0.103±0.007 0.098±0.011 The specific yield of sclerotin (%) 0.067±0.005 0.140±0.012 0.060±0.004 0.058±0.005 Example 16 Mycelium content (mg/g) 131.06±8.10 190.51±7.92 182.70±5.53 170.25±2.69 Hardhair content (mg/g) 0.082±0.005 0.272±0.028 0.118±0.017 0.092±0.008 The specific yield of sclerotin (%) 0.063±0.004 0.143±0.010 0.065±0.007 0.054±0.003

<Analysis of the influence of the grain type of the culture material on the yield of sclerostin>

In this experiment, the above-mentioned Example 13 and Example 17 to Example 23 were used to conduct experiments to analyze the sclerostin content and The specific yield of sclerostin of mycelium, and the relevant steps and details of the solid-state fermentation method of sclerostin of the present invention, please refer to the above, and will not be repeated here.

Please refer to Table 14, which shows the mycelium content, sclerotin content and the sclerotin ratio of the mycelium of the fermented cultures of Example 13, Example 17 to Example 23 after culturing for 5 weeks to 8 weeks. Rate. As can be seen from the results of Table 14, the sclerotin content of Example 13, Example 17 to Example 23 has a good sclerotin content and the sclerotin ratio of the mycelium when culturing 6 weeks and 7 weeks. The rate performance, wherein the black rice of Example 18 and the rye of Example 21 were the best, and the sclerotin specific yield of its mycelium reached more than 0.610% when cultured for 6 weeks, showing that brown rice, millet, Black rice, rye, oats, buckwheat, wheat and barley can all be used as the culture materials of the solid-state fermentation method for sclerostin of the present invention, and can effectively generate and improve the content of sclerostin in the fermentation culture, so that it has Growth potential of healthcare and medical-related markets. Table 14 Cultivation time (weeks) 5 6 7 8 Example 13 Mycelium content (mg/g) 145.20±15.10 165.42±15.12 175.50±18.25 200.32±18.12 Hardhair content (mg/g) 0.093±0.006 0.350±0.014 0.227±0.014 0.104±0.004 The specific yield of sclerotin (%) 0.064±0.004 0.212±0.009 0.129±0.012 0.052±0.002 Example 17 Mycelium content (mg/g) 30.65±0.05 75.13±3.75 45.40±1.24 48.85±2.76 Hardhair content (mg/g) 0.047±0.007 0.083±0.003 0.058±0.004 0.111±0.008 The specific yield of sclerotin (%) 0.153±0.002 0.111±0.004 0.129±0.008 0.227±0.012 Example 18 Mycelium content (mg/g) 52.76±5.01 110.41±3.67 114.60±6.42 91.38±5.04 Hardhair content (mg/g) 0.084±0.001 0.819±0.018 0.558±0.012 0.167±0.021 The specific yield of sclerotin (%) 0.159±0.012 0.742±0.017 0.487±0.024 0.183±0.036 Example 19 Mycelium content (mg/g) 106.37±2.16 180.57±0.37 140.64±0.66 120.56±1.47 Hardhair content (mg/g) 0.060±0.001 0.138±0.012 0.029±0.008 0.023±0.007 The specific yield of sclerotin (%) 0.057±0.002 0.077±0.003 0.021±0.002 0.019±0.001 Example 20 Mycelium content (mg/g) 75.38±0.544 92.19±0.20 155.01±1.24 125.51±0.72 Hardhair content (mg/g) 0.098±0.010 0.122±0.017 0.199±0.015 0.054±0.010 The specific yield of sclerotin (%) 0.131±0.003 0.133±0.006 0.128±0.003 0.043±0.003 Example 21 Mycelium content (mg/g) 110.39±6.30 180.45±10.22 195.87±10.74 200.91±10.82 Hardhair content (mg/g) 0.303±0.017 1.107±0.050 0.599±0.021 0.185±0.012 The specific yield of sclerotin (%) 0.274±0.020 0.613±0.035 0.306±0.010 0.092±0.005 Example 22 Mycelium content (mg/g) 95.58±5.13 155.51±10.28 147.62±15.24 143.88±10.24 Hardhair content (mg/g) 0.331±0.025 0.145±0.011 0.050±0.011 0.027±0.009 The specific yield of sclerotin (%) 0.346±0.028 0.093±0.007 0.034±0.004 0.019±0.003 Example 23 Mycelium content (mg/g) 90.65±6.05 135.72±12.70 180.71±11.26 181.89±12.31 Hardhair content (mg/g) 0.080±0.012 0.354±0.024 0.815±0.060 0.342±0.041 The specific yield of sclerotin (%) 0.089±0.007 0.261±0.015 0.451±0.035 0.188±0.014

<Analysis of the influence of the carbon source of the culture material on the yield of sclerotin>

In this experiment, the above-mentioned Example 24 to Example 26 and Comparative Example 10 to Comparative Example 11 were used to conduct experiments to analyze the fermentation culture obtained by the solid-state fermentation method of sclerostin of the present invention when the culture material contains different kinds of carbon sources The sclerostin content of the material and the sclerostin specific yield of the mycelium, and the relevant steps and details of the solid-state fermentation method of sclerostin of the present invention, please refer to the above, and will not be repeated here.

Please refer to Table 15, which series shows the mycelium content, sclerostin content and the hardness of the mycelium of the fermented cultures from Examples 24 to 26 and Comparative Examples 10 to 11 when cultured for 5 to 8 weeks. Gross ratio yield. As can be seen from the results in Table 15, the sclerotin content was the highest with Example 26 when cultured for 7 weeks, and its sclerotin content was 0.290 mg/g, and the sclerotin content of Example 13 was also the highest when cultured for 7 weeks. It can reach 0.270 mg/g, and the bristle content of Examples 24 to 26 is better than that of Comparative Example 10 and Comparative Example 11, indicating that glucose, mannose and sucrose can all be used as the bristle of the present invention It is the carbon source of the raw material for the solid-state fermentation of vegetarian food, and has related market potential. Table 15 Cultivation time (weeks) 5 6 7 8 Example 24 Mycelium content (mg/g) 105.63±1.24 184.07±18.12 164.15±1.76 163.81±2.45 Hardhair content (mg/g) 0.031±0.005 0.105±0.011 0.106±0.005 0.039±0.001 The specific yield of sclerotin (%) 0.029±0.003 0.057±0.005 0.065±0.002 0.024±0.000 Example 25 Mycelium content (mg/g) 129.93±3.32 188.08±5.70 189.90±4.70 192.47±1.64 Hardhair content (mg/g) 0.041±0.004 0.112±0.015 0.270±0.010 0.141±0.013 The specific yield of sclerotin (%) 0.032±0.002 0.060±0.005 0.142±0.005 0.073±0.004 Example 26 Mycelium content (mg/g) 120.80±5.35 160.09±3.42 158.64±1.66 140.63±3.97 Hardhair content (mg/g) 0.030±0.002 0.107±0.020 0.290±0.002 0.155±0.015 The specific yield of sclerotin (%) 0.025±0.001 0.067±0.009 0.183±0.005 0.110±0.008 Comparative Example 10 Mycelium content (mg/g) 125.78±4.90 167.78±3.81 165.43±1.75 138.09±4.70 Hardhair content (mg/g) 0.032±0.005 0.090±0.005 0.083±0.008 0.011±0.000 The specific yield of sclerotin (%) 0.025±0.003 0.054±0.002 0.050±0.003 0.008±0.000 Comparative Example 11 Mycelium content (mg/g) 110.40±10.35 190.60±5.00 154.44±7.04 160.88±1.10 Hardhair content (mg/g) 0.023±0.005 0.060±0.004 0.022±0.000 0.009±0.000 The specific yield of sclerotin (%) 0.021±0.003 0.032±0.002 0.014±0.001 0.006±0.000

<Analysis of the influence of the nitrogen source of the culture material on the yield of sclerotin>

This experiment is based on the aforementioned Example 27 to Example 29 and Comparative Example 12 to analyze the bristles of the fermented culture obtained by the solid-state fermentation method of bristle of the present invention when the culture material contains different kinds of nitrogen sources The content of sclerostin and the specific yield of sclerostin of mycelium, and the relevant steps and details of the solid-state fermentation method of sclerostin of the present invention, please refer to the above, and will not be repeated here.

Please refer to Table 16, which shows the mycelium content, sclerotin content and the sclerotin ratio of the mycelium of the fermented cultures of Example 27 to Example 29 and Comparative Example 12 in 5 to 8 weeks of culture. Rate. As can be seen from the results in Table 16, the sclerotin content was the highest with Example 28 when culturing for 7 weeks, and its sclerotin content was 0.233 mg/g, and the sclerotin specific yield of its mycelium could also reach 0.134 %, and compared with Comparative Example 12, the sclerotin content of Examples 27 to 29 are all better, indicating that yeast extract, malt extract and egg white can be used as part of the solid-state fermentation method of sclerostin of the present invention. Nitrogen source for cultivating raw materials, and has related market potential. Table 16 Cultivation time (weeks) 5 6 7 8 Example 27 Mycelium content (mg/g) 125.79±1.82 166.31±4.41 178.59±2.41 167.77±2.26 Hardhair content (mg/g) 0.055±0.006 0.113±0.002 0.195±0.006 0.081±0.001 The specific yield of sclerotin (%) 0.044±0.003 0.068±0.001 0.110±0.003 0.048±0.005 Example 28 Mycelium content (mg/g) 115.32±1.74 189.89±3.44 174.53±5.06 164.95±3.20 Hardhair content (mg/g) 0.063±0.002 0.109±0.008 0.233±0.009 0.103±0.008 The specific yield of sclerotin (%) 0.055±0.001 0.057±0.003 0.134±0.004 0.062±0.004 Example 29 Mycelium content (mg/g) 108.72±2.11 176.73±3.03 181.92±5.03 147.34±4.91 Hardhair content (mg/g) 0.085±0.005 0.134±0.006 0.087±0.003 0.036±0.000 The specific yield of sclerotin (%) 0.079±0.005 0.076±0.003 0.048±0.002 0.024±0.001 Comparative Example 12 Mycelium content (mg/g) 103.49±6.35 154.18±5.65 143.81±2.88 139.28±1.75 Hardhair content (mg/g) 0.013±0.002 0.018±0.000 0.010±0.001 0.017±0.001 The specific yield of sclerotin (%) 0.013±0.001 0.012±0.000 0.007±0.000 0.012±0.000

<Analysis of the effect of strain activation time on the yield of sclerostin>

In this experiment, the above-mentioned Example 13, Example 30 and Example 31 were used to analyze the solid-state fermentation method of sclerostin of the present invention after treatment with different bacterial species activation time. The sclerostin content of the fermented culture obtained during the experiment and the sclerostin specific yield of the mycelium, and the relevant steps and details of the solid-state fermentation method of sclerostin of the present invention, please refer to the above, and will not be repeated here. Repeat.

Please refer to Table 17, which shows the mycelium content, sclerotin content and the sclerotin ratio of the mycelium of the fermented cultures of Example 13, Example 30 and Example 31 in 5 to 8 weeks of culture. Rate. As can be seen from the results in Table 17, the sclerotin contents of Example 13, Example 30 and Example 31 were 0.350 mg/g, 0.219 mg/g and 0.209 mg/g respectively when the culture time was 6 weeks, and the bacteria were The specific yields of sclerotin of silk are 0.212%, 0.129% and 0.119% respectively, showing that the solid-state fermentation method of sclerotin of the present invention is carried out by the S. schizophrenia of the present invention after being treated with different strains of activation time. The solid-state fermentation of sclerostin can effectively generate sclerostin, which has the potential for the development of health care and medical related markets. Table 17 Cultivation time (weeks) 5 6 7 8 Example 13 Mycelium content (mg/g) 145.20±15.10 165.42±15.12 175.50±18.25 200.32±18.12 Hardhair content (mg/g) 0.093±0.006 0.350±0.014 0.227±0.014 0.104±0.004 The specific yield of sclerotin (%) 0.064±0.004 0.212±0.009 0.129±0.012 0.052±0.002 Example 30 Mycelium content (mg/g) 152.28±10.24 170.12±18.60 189.55±12.36 198.11±20.15 Hardhair content (mg/g) 0.103±0.002 0.219±0.024 0.116±0.008 0.076±0.000 The specific yield of sclerotin (%) 0.068±0.001 0.129±0.014 0.061±0.004 0.038±0.004 Example 31 Mycelium content (mg/g) 160.20±9.17 175.25±20.33 185.30±10.16 196.20±14.30 Hardhair content (mg/g) 0.074±0.002 0.209±0.015 0.082±0.002 0.057±0.002 The specific yield of sclerotin (%) 0.046±0.001 0.119±0.009 0.044±0.001 0.029±0.002

<Analysis of the influence of the inoculation amount of the Xyloporus schizophrenia bacterial solution in the culture material on the yield of sclerotin>

In this experiment, the above-mentioned Example 13, Example 32 and Example 33 are used to conduct experiments to analyze the solid-state fermentation method of sclerostin of the present invention when the inoculum of Xyloporus schizophylla of the present invention is different in the culture material The sclerostin content of the obtained fermented culture and the sclerostin specific yield of the mycelium, and the relevant steps and details of the solid-state fermentation method of sclerostin of the present invention, please refer to the above, and will not be repeated here.

Please refer to Table 18, which serially shows the mycelium content, the sclerotin content and the sclerotin ratio of the mycelium of the fermented cultures of Example 13, Example 32 and Example 33 after culturing for 5 to 8 weeks. Rate. It can be seen from the results in Table 18 that the sclerotin contents of Example 13, Example 32 and Example 33 were 0.227 mg/g, 0.219 mg/g and 0.149 mg/g respectively when the culture time was 7 weeks, showing that the The solid-state fermentation method of sclerostin of the invention can effectively generate sclerostin when the inoculum of the culturing material is 5%, 10% and 15% of the inoculum of the present invention, wherein the inoculum is 10%. When the sclerostin content is optimal, the solid-state fermentation method of sclerostin of the present invention can be used to mass-produce sufficient sclerostin when the low inoculation amount of the present invention's sclerostin Development potential of the relevant market Table 18 Cultivation time (weeks) 5 6 7 8 Example 13 Mycelium content (mg/g) 145.20±15.10 165.42±15.12 175.50±18.25 200.32±18.12 Hardhair content (mg/g) 0.093±0.006 0.350±0.014 0.227±0.014 0.104±0.004 The specific yield of sclerotin (%) 0.064±0.004 0.212±0.009 0.129±0.012 0.052±0.002 Example 32 Mycelium content (mg/g) 127.53±2.70 169.58±5.72 185.85±6.43 161.46±1.50 Hardhair content (mg/g) 0.059±0.000 0.082±0.008 0.219±0.021 0.088±0.011 The specific yield of sclerotin (%) 0.046±0.002 0.048±0.005 0.118±0.011 0.055±0.007 Example 33 Mycelium content (mg/g) 140.46±4.14 176.26±3.66 201.10±13.06 193.02±5.82 Hardhair content (mg/g) 0.102±0.001 0.127±0.002 0.149±0.040 0.126±0.003 The specific yield of sclerotin (%) 0.073±0.001 0.072±0.001 0.074±0.020 0.065±0.002

<Analysis of the influence of the liquid medium type of Xyloporus schizophrenia of the present invention on the yield of sclerotin>

In this experiment, the above-mentioned Example 13, Example 34 and Example 35 were used to conduct experiments to analyze the solid-state fermentation method of sclerostin of the present invention when culturing Xyloporus schizophrenia of the present invention activated with different liquid media The sclerostin content of the obtained fermented culture and the sclerostin specific yield of the mycelium, and the relevant steps and details of the solid-state fermentation method of sclerostin of the present invention, please refer to the above, and will not be repeated here.

Please refer to Table 19, which shows the mycelium content, the sclerotin content and the sclerotin ratio of the mycelium of the fermented cultures of Example 13, Example 34 and Example 35 for 5 to 8 weeks of culture. Rate. It can be seen from the results in Table 19 that the sclerotin contents of Example 13, Example 34 and Example 35 were 0.227 mg/g, 0.127 mg/g and 0.191 mg/g respectively when the culture time was 7 weeks, showing that the The solid-state fermentation method of sclerostin of the invention can effectively generate sclerotin when using the PDB medium, MEB medium and YGB medium to activate the schizophrenia of the present invention for culturing, so that the solid-state sclerotin of the present invention can be effectively produced. Fermentation methods are more widely applicable and have potential for health and medical related markets. Table 19 Cultivation time (weeks) 5 6 7 8 Example 13 Mycelium content (mg/g) 145.20±15.10 165.42±15.12 175.50±18.25 200.32±18.12 Hardhair content (mg/g) 0.093±0.006 0.350±0.014 0.227±0.014 0.104±0.004 The specific yield of sclerotin (%) 0.064±0.004 0.212±0.009 0.129±0.012 0.052±0.002 Example 34 Mycelium content (mg/g) 108.48±9.37 154.35±10.18 170.20±12.50 178.24±10.33 Hardhair content (mg/g) 0.079±0.009 0.113±0.002 0.127±0.014 0.238±0.070 The specific yield of sclerotin (%) 0.073±0.008 0.073±0.001 0.075±0.008 0.134±0.039 Example 35 Mycelium content (mg/g) 105.24±8.35 144.27±8.25 165.24±10.50 168.10±15.35 Hardhair content (mg/g) 0.070±0.010 0.098±0.005 0.191±0.003 0.082±0.001 The specific yield of sclerotin (%) 0.067±0.010 0.068±0.003 0.116±0.003 0.049±0.001

To sum up, the solid-state fermentation method of sclerostin of the present invention conducts solid-state fermentation through the Schizophrenia schizophrenia of the present invention, and can be matched with suitable initial pH value of culture raw materials, inoculum amount, and other culture conditions and steps as required. , or by adding metal ion additives KH ₂ PO ₄ or ZnCO ₃ to the culture raw materials, changing the temperature difference conditions of solid-state fermentation or changing the light conditions of solid-state fermentation, it will be effective to generate sclerostin in the fermentation culture, and then make this The invented solid-state fermentation method of sclerostin can quickly and efficiently generate and improve the yield of sclerostin, and has the development potential of the relevant market.

However, the present invention has been disclosed as above in an embodiment, but it is not intended to limit the present invention. Anyone who is familiar with the art can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection of the present invention The scope shall be determined by the scope of the appended patent application.

100, 200, 300, 400: Solid-state fermentation method of sclerostin 110, 120, 130, 140, 210, 220, 230, 240, 310, 320, 330, 340, 410, 420, 430, 440: Steps 241: The first thermophilic culture step 242: Second thermophilic culture step 341: First Light Cultivation Step 342: Second Light Cultivation Step

FIG. 1 is a flow chart showing the steps of a solid-state fermentation method for sclerostin according to an embodiment of the present invention; Fig. 2 is a flow chart showing the steps of the solid-state fermentation method of sclerostin according to another embodiment of the present invention; Fig. 3 is a flow chart showing the steps of the solid-state fermentation method of sclerostin according to another embodiment of the present invention; FIG. 4 is a flow chart showing the steps of a solid-state fermentation method for sclerostin according to still another embodiment of the present invention; The 5th figure is the mycelium growth situation of the schizophrenia of the present invention cultivated in brown rice medium under different temperature difference conditions; Fig. 6 is the mycelial growth situation of Xyloporus cleft hoof of the present invention cultivated in rye culture medium under different temperature difference conditions; Fig. 7 is the mycelium growth situation of Xyloporus cleft hoof of the present invention cultivated in brown rice medium under different light conditions; and Figure 8 shows the mycelium growth of Xyloporus schizophrenia in rye culture medium under different light conditions.

Domestic storage information (please note in the order of storage institution, date and number) Food Industry Development Institute September 20, 108 Deposit number: BCRC 930211

100: Solid-state fermentation method of sclerostin

110, 120, 130, 140: Steps

Claims (14)

A solid-state fermentation method for hispidin, comprising the steps of: providing a culture material, wherein the culture material comprises a grain culture medium, the grain culture medium comprises a brown rice, a rye or a combination thereof, a The water content is 50% to 70% by weight, and one of the initial pH values of the culture raw materials is 5.5 to 6.5; a bacterial species activation step is carried out, which is to inoculate a Phellinus linteus into a liquid culture medium In and cultivate an activation time, so as to obtain a Xylophoa spp. bacterium liquid, wherein this activation time is 2 weeks, and the deposit number of this Xylophoa spp is BCRC 930211; Carry out an inoculation step, it is linked to this cultivation Inoculate this Xylomicronia bacterium liquid in the raw material, to obtain a mixture, wherein this Xylem inoculum is 5%～15% weight ratio in one of this culturing raw material inoculation amount; And carry out a cultivating step, its The mixture is cultured in the dark for 6 weeks to 8 weeks at a culture temperature to obtain a fermentation culture, and the fermentation culture comprises a sclerostin; wherein, the culture step comprises: a first temperature difference culture step, wherein The culturing temperature of the first temperature difference culturing step is 20°C to 25°C; and a second temperature difference culturing step, wherein the culture temperature of the second temperature difference culturing step is 16°C to 19°C; wherein, the mixture is in the The first thermophilic culture step is cultured until one of the hyphae of the Xylem indica is completely entangled with the mixture. The solid state fermentation method of sclerostin according to claim 1, wherein the initial pH value of the culture material is 5.5. The solid-state fermentation method of sclerostin according to claim 1, wherein the activation time is 1 week. The solid-state fermentation method of sclerostin as claimed in claim 1, wherein the inoculation amount of the Xyloporus spp. in the culture raw material is 10% by weight. The method for solid-state fermentation of sclerostin according to claim 1, wherein the liquid medium is PDB medium (Potato dextrose broth), MEB medium (Malt extract broth) or YGB medium (Yeast extract glucose broth). The solid state fermentation method of sclerostin according to claim 1, wherein the culture material further comprises a carbon source, and the carbon source is glucose, mannose or sucrose. The method for solid-state fermentation of sclerostin according to claim 1, wherein the culture material further comprises a nitrogen source, and the nitrogen source is yeast extract, malt extract or protein gluten. A kind of solid-state fermentation method of sclerostin, comprising the steps: A culture material is provided, wherein the culture material comprises a grain medium, the grain medium comprises brown rice, a rye or a combination thereof, a water content of the culture material is 50% to 70% by weight, and one of the culture materials is The initial pH value is 5.5 to 6.5; a bacterial classification activation step is carried out, which is to inoculate a Xylomicronia in a liquid medium and cultivate for an activation time to obtain a Xylomicron bacteria liquid, wherein the activation The time is 2 weeks, and the deposit number of the Xylomya cleft hoof is BCRC 930211; an inoculation step is carried out, and it is inoculated with the Xylophyllanum spp. bacterial liquid in the culture material to obtain a mixture, wherein the S. One of the inoculum amounts of the pore bacteria liquid in the culture material is 5% to 15% by weight; and a culture step is carried out, wherein the mixture is cultured at a culture temperature for 6 weeks to 8 weeks to obtain a fermentation culture, And the fermentation culture comprises a sclerostin; wherein, the culture temperature is 20°C to 25°C, and the culture step comprises: a first illumination culture step, which is to culture the mixture in the dark; and a second illumination culture step, wherein the second light culture step is carried out under a photoperiod of 12 hours of light and 12 hours of darkness, and one of the light illumination of the second light culture step is 450 lux ~ 550 lux; wherein, the mixture is in the first The light culture step is cultured until one of the hyphae of the Xyloporus schizophrenia is completely entangled with the mixture. The solid state fermentation method of sclerostin as claimed in claim 8, Wherein the initial pH value of the culture material is 5.5. The solid-state fermentation method of sclerostin as claimed in claim 8, wherein the inoculation amount of the Xyloporus spp. in the culture raw material is 10% by weight. The solid-state fermentation method of sclerostin according to claim 8, wherein the activation time is 1 week. The method for solid-state fermentation of sclerostin according to claim 8, wherein the liquid medium is PDB medium (Potato dextrose broth), MEB medium (Malt extract broth) or YGB medium (Yeast extract glucose broth). The solid-state fermentation method of sclerostin according to claim 8, wherein the culture material further comprises a carbon source, and the carbon source is glucose, mannose or sucrose. The method for solid-state fermentation of sclerostin according to claim 8, wherein the culture material further comprises a nitrogen source, and the nitrogen source is yeast extract, malt extract or protein gluten.

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