Method for liquid fermentation of red ganoderma

Abstract

The present application relates to a kind of ganoderma lucidum liquid fermentation method, specifically to a kind of method for improving ganoderma triterpenes content in liquid fermentation by adding specific components to fermentation medium.The method can significantly improve the content of ganoderma triterpenes in fermentation broth, and has synergistic effect.

Description

Technical Field

[0001] This invention relates to a method for liquid fermentation of Ganoderma lucidum, and more specifically, to a method for increasing the content of Ganoderma lucidum triterpenes in liquid fermentation by adding specific components to the fermentation medium. Background Technology

[0002] Ganoderma lucidum (Leyss. ex Fr.) Karst, belonging to the Polyporaceae family of fungi, has mycelium, fruiting bodies, and spores that can be used for health maintenance or medicinal purposes. Ganoderma triterpenes are a class of triterpenoid compounds found in Ganoderma lucidum, belonging to highly oxidized lanosterane derivatives, and are one of the main chemical and pharmacologically active components of Ganoderma lucidum. The ganoderic triterpenes contained in Ganoderma lucidum possess particularly significant physiological activity.

[0003] Therefore, a simple and effective method is still needed to increase the content of Ganoderma triterpenes in fermentation broth for use in the production of Ganoderma-related products. Summary of the Invention

[0004] This invention provides a method for liquid fermentation of Ganoderma lucidum, comprising adding allantoin or further adding vitamin C phosphate to the fermentation medium. This method can significantly increase the content of Ganoderma lucidum triterpenes in the fermentation broth and has a synergistic effect.

[0005] This invention has found that adding allantoin to the fermentation medium can increase the content of Ganoderma lucidum triterpenes by 10-18%. Specifically, the amount of allantoin added to the fermentation medium is 0.1-0.25% (w / v), for example, 0.1%, 0.15%, 0.2%, and 0.25%. If the amount of allantoin added is less than 0.1%, the effect of increasing the content of Ganoderma lucidum triterpenes is not significant; if it exceeds 0.25%, it does not help to further increase the content of Ganoderma lucidum triterpenes.

[0006] Further research revealed that adding allantoin followed by vitamin C phosphate to the fermentation medium could further increase the content of Ganoderma lucidum triterpenes by 5-8%. The addition amount of vitamin C phosphate in the fermentation medium was 0.05-0.1% (w / v), specifically 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, and 0.1%. However, adding vitamin C phosphate alone to the fermentation medium had little effect on increasing the content of Ganoderma lucidum triterpenes, meaning there was essentially no increase. This indicates that allantoin and vitamin C phosphate have a synergistic effect in increasing the content of Ganoderma lucidum triterpenes.

[0007] 35% vitamin C phosphate was used in this article.

[0008] In some preferred embodiments, the fermentation medium used is: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg VB1 (vitamin B1), 0.5g casein; with water added to 1000ml.

[0009] As mentioned above, the specific amounts of allantoin and vitamin C phosphate added to the fermentation medium are crucial for achieving the desired effect. Adding allantoin in the range of 0.1-0.25% (w / v) can increase the content of Ganoderma lucidum triterpenes by 10-18%, while adding vitamin C phosphate in the range of 0.05-0.1% (w / v) can further increase the content of Ganoderma lucidum triterpenes by 5-8%. The combination of allantoin and vitamin C phosphate can have a synergistic effect on increasing the content of Ganoderma lucidum triterpenes.

[0010] In some specific examples, the fermentation was carried out in a 250ml Erlenmeyer flask containing 75ml of fermentation medium, with an inoculum size of 10%, a fermentation time of 96 hours, a temperature of 28°C, and a stirring speed of 120 rpm. The content of Ganoderma triterpenes in the fermentation broth can be determined using conventional methods in the art.

[0011] Based on the above research, the present invention also provides a fermentation culture medium composition, which is made by adding 0.1-0.25% (w / v) of allantoin to the above fermentation culture medium. Another preferred embodiment is that 0.05-0.1% (w / v) of vitamin C phosphate is also added to the above fermentation culture medium.

[0012] A fermentation medium composition has the following formulation: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, 0.5g casein, and 1-2.5g allantoin; add water to 1000ml. Optionally, the fermentation medium composition further contains 0.5-1g of vitamin C phosphate (i.e., per 1000ml).

[0013] This invention does not have any particular limitation on the Ganoderma lucidum strain used; all known Ganoderma lucidum strains can achieve essentially the same fermentation effect.

[0014] The fermentation medium of the present invention can be prepared using conventional methods in the art, and there are no particular limitations.

[0015] 1. A method for liquid fermentation of Ganoderma lucidum, comprising adding allantoin to the fermentation culture medium.

[0016] 2. According to the method described in 1, the amount of allantoin added to the fermentation medium is 0.1-0.25% (w / v).

[0017] 3. According to the method described in 1, the amount of allantoin added to the fermentation medium is 0.1%, 0.15%, 0.2% or 0.25%.

[0018] 4. The method according to 1 further includes adding vitamin C phosphate to the fermentation medium.

[0019] 5. According to the method described in 4, the amount of vitamin C phosphate added to the fermentation medium is 0.05-0.1% (w / v).

[0020] 6. According to the method described in 4, the amount of vitamin C phosphate added to the fermentation medium is 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1%.

[0021] 7. According to the aforementioned method, the fermentation medium used is: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, and 0.5g casein; add water to 1000ml.

[0022] 8. The method described above includes providing a fermentation medium; adding allantoin to the fermentation medium, such that the amount of allantoin added to the fermentation medium is 0.1-0.25% (w / v); and inoculating with Ganoderma lucidum strains for fermentation.

[0023] 9. The method according to 8 further includes adding vitamin C phosphate to the fermentation medium, such that the amount of vitamin C phosphate added to the fermentation medium is 0.05-0.1% (w / v); and then inoculating with Ganoderma lucidum for fermentation.

[0024] 10. A fermentation culture medium composition, with the following formula: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, 0.5g casein, 1-2.5g allantoin; add water to 1000ml.

[0025] 11. The fermentation medium composition according to 10, wherein the fermentation medium composition further contains 0.5-1g of vitamin C phosphate (i.e., per 1000ml).

[0026] This invention provides an improved method for producing Ganoderma lucidum triterpenes, in which allantoin and vitamin C phosphate are added to the fermentation medium, which has a synergistic effect on increasing the content of Ganoderma lucidum triterpenes. Detailed Implementation

[0027] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention.

[0028] The fermentation conditions were as follows: 75 mL of fermentation medium in a 250 mL Erlenmeyer flask, 10% inoculum, 96 h culture time, 28 °C culture temperature, and 120 rpm rotation speed.

[0029] The Ganoderma lucidum used below is Hanzhi No. 3 (purchased commercially).

[0030] The following method is used to detect Ganoderma lucidum triterpenes: The fermentation broth is centrifuged at 5000 r / min for 15 minutes to obtain the supernatant, which is then concentrated. Five times the volume of industrial alcohol is added, and after precipitation, 200 mL of the supernatant is taken. After evaporating the water, it is extracted by reflux with 500 mL of 95% ethanol solution for 2 hours. The extract is then extracted with n-butanol at 50℃ with shaking for 3 hours. The n-butanol is evaporated at 60℃ to obtain the extract. After dissolving it in methanol, it is used as a sample. After a colorimetric reaction, the absorbance is measured at 250 nm, and the content of Ganoderma lucidum triterpenes is calculated.

[0031] Example 1 This embodiment provides a method for liquid fermentation of Ganoderma lucidum, including inoculating a culture of Ganoderma lucidum fungi into a fermentation medium for fermentation. After fermentation, the concentration of Ganoderma lucidum triterpenes in the fermentation broth was found to be 344.9 mg / L.

[0032] Fermentation medium: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, 0.5g casein, 1g allantoin; add water to 1000ml.

[0033] Example 2 This embodiment provides a method for liquid fermentation of Ganoderma lucidum, including inoculating a culture of Ganoderma lucidum fungi into a fermentation medium for fermentation. After fermentation, the concentration of Ganoderma lucidum triterpenes in the fermentation broth was found to be 371.1 mg / L.

[0034] Fermentation medium: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, 0.5g casein, 2.5g allantoin; add water to 1000ml.

[0035] Example 3 This embodiment provides a method for liquid fermentation of Ganoderma lucidum, including inoculating a culture of Ganoderma lucidum fungi into a fermentation medium for fermentation. After fermentation, the concentration of Ganoderma lucidum triterpenes in the fermentation broth was found to be 388.5 mg / L.

[0036] Fermentation medium: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, 0.5g casein, 1g allantoin, 1g vitamin C phosphate; add water to 1000ml.

[0037] Example 4 This embodiment provides a method for liquid fermentation of Ganoderma lucidum, including inoculating a culture of Ganoderma lucidum fungi into a fermentation medium for fermentation. After fermentation, the concentration of Ganoderma lucidum triterpenes in the fermentation broth was found to be 403.4 mg / L.

[0038] Fermentation medium: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, 0.5g casein, 2.5g allantoin, 0.8g vitamin C phosphate; add water to 1000ml.

[0039] Comparative Example 1 This comparative example provides a method for liquid fermentation of Ganoderma lucidum, including inoculating a culture of Ganoderma lucidum fungi into a fermentation medium for fermentation. After fermentation, the concentration of Ganoderma lucidum triterpenes in the fermentation broth was found to be 315 mg / L.

[0040] Fermentation medium: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, 0.5g casein; add water to 1000ml.

[0041] Comparative Example 2 This comparative example provides a method for liquid fermentation of Ganoderma lucidum, including inoculating a culture of Ganoderma lucidum fungi into a fermentation medium for fermentation. After fermentation, the concentration of Ganoderma lucidum triterpenes in the fermentation broth was found to be 318.2 mg / L.

[0042] Fermentation medium: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, 0.5g casein, 1g vitamin C phosphate; add water to 1000ml.

[0043] It should be understood that the embodiments described above are for illustrative purposes only, and modifications and variations may be made without departing from the scope of the invention as defined in the appended claims.

Claims

1. A method for liquid fermentation of Ganoderma lucidum, characterized in that, include: Provide fermentation culture medium; Allantoin and vitamin C phosphate are added to the fermentation medium, with allantoin added at a w / v level of 0.1-0.25% and vitamin C phosphate added at a w / v level of 0.05-0.1%. Ganoderma lucidum is inoculated for fermentation. The fermentation medium consists of: 25g wheat bran, 30g glucose, 1.5g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.6g diammonium hydrogen phosphate, 10mg vitamin B1, and 0.5g casein; water is added to 1000ml.

2. The method according to claim 1, wherein the amount of allantoin added to the fermentation medium is 0.1%, 0.15%, 0.2%, or 0.25%.

3. The method according to claim 1, wherein the amount of vitamin C phosphate added to the fermentation medium is 0.05%, 0.06%, 0.07%, 0.08%, 0.09% or 0.1%.