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Method for producing 4-Acetylantroquinonol B through fermentation

A technology for liquid fermentation and strain, applied in the field of fermented production of 4-acetyl androquinol B, can solve the problems of low yield, insufficient fruiting bodies of Antrodia camphorata to supply market demand, shortage of camphor tree resources and the like

Active Publication Date: 2016-04-06
厦门北化生物产业研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to the strict host characteristics of Antrodia camphorata and the scarcity of natural output, and its only host camphor tree resources are also very scarce, the fruiting body of Antrodia camphorata is far from enough to supply market demand and the price is very expensive
The use of modern biotechnology to artificially cultivate Antrodia camphorata to obtain bacterial products is an effective solution to the contradiction between supply and demand of Antrodia camphorata. Yu-WeiLin et al. have identified 4-AcetylantroquinonolB from liquid cultured Antrodia camphorata bacteria, but its yield is low; Chien-ChiChiang etc. adopt the mode of liquid fermentation, effectively improved the content of 4-AcetylantroquinonolB in the bacterium of liquid state fermentation by adding inducer, finally its yield reaches 0.8mg / g; Therefore obtain the content of 4-AcetylantroquinonolB by liquid fermentation method The research is still in its infancy, and the research on the production of 4-AcetylantroquinonolB by liquid fermentation without adding any inducers or prerequisites has not been reported yet.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Taking the fermentation of a triangular flask with a liquid volume of 250mL as an example, the operation process of Antrodia camphorata liquid seed cultivation is as follows:

[0028] 1) preparation of spores of Antrodia camphorata:

[0029] Using the inoculation method of digging blocks, inoculate Antrodia antrodia into a new solid medium. The composition of the solid medium is: glucose 20g / L, peptone 2.5g / L, malt extract powder 20g / L, agar 25g / L, cultured in the dark After 20-25 days, a new circular colony is formed with a diameter of about 4-5cm. The center of the colony is orange and the edge is white. Store it at 4°C for later use.

[0030] 2) preparation of Antrodia spore suspension:

[0031] New colonies after expansion can be used to prepare spore suspension. After the strains stored at 4°C were placed at room temperature for 1-2 hours, 10-15mL of sterile water was added under aseptic conditions, and the spores on the surface of the colonies were gently scrape...

Embodiment 2

[0036] Taking the fermentation of a triangular flask with a liquid volume of 250mL as an example, the operation process of producing 4-AcetylantroquinonolB by liquid fermentation of Antrodia camphorata is as follows:

[0037] 1) The first stage of fermentation culture:

[0038] With the obtained Antrodia camphorata globule in embodiment 1 as seed liquid, inoculate in the first-stage liquid fermentation medium with 15% inoculum size, liquid loading is 50mL / 250mL, and liquid fermentation medium consists of: glucose 15g / L, Peptone 5g / L, K 2 HPO 4 0.5g / L, MgSO 4 ·7H 2 O0.1g / L, malt extract powder 15g / L, phenol 0.05g / L. Placed in a constant temperature shaker at 120 rpm at 22°C for 8 days.

[0039] 2) The second stage fermentation culture:

[0040] 1.5 g of glucose was added to the fermentation medium, and cultured in a constant temperature shaker at 180 rpm at 28° C. for 5 days.

[0041] 3) Extraction and determination of 4-AcetylantroquinonolB in bacteria:

[0042] After th...

Embodiment 3

[0045] Taking the fermentation of a triangular flask with a liquid volume of 250mL as an example, the operation process of producing 4-AcetylantroquinonolB by liquid fermentation of Antrodia camphorata is as follows:

[0046] 1) The first stage of fermentation culture:

[0047] With the obtained Antrodia camphorata globule in embodiment 1 as seed liquid, inoculate in the first-stage fermentation medium with 10% inoculum size, liquid load is 50mL / 250mL, and fermentation medium consists of: glucose 18g / L, peptone 6g / L,K 2 HPO 4 0.8g / L, MgSO 4 ·7H 2 O0.5g / L, malt extract powder 18g / L, phenol 0.1g / L. Placed in a constant temperature shaker at 100 rpm at 20°C for 10 days.

[0048] 2) The second stage fermentation culture:

[0049] 0.5 g of glucose was added to the fermentation medium, and cultured in a constant temperature shaker at 180 rpm at 27° C. for 5 days.

[0050] 3) Extraction and determination of 4-AcetylantroquinonolB in bacteria:

[0051] Take a certain amount o...

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Abstract

The invention belongs to the technical field of fermentation, and particularly relates to a method for producing 4-Acetylantroquinonol B through fermentation. Antrodia camphorate is a fungus growing on Taiwan rotten Cinnamomum kanehirai Hay. in an obligatory parasitism mode, fruiting bodies of antrodia camphorate contain various natural active substances, 4-Acetylantroquinonol B is a substance which is separated out of the fruiting bodies of antrodia camphorate in recent years and has a good anti-cancer effect, but at present, research on antrodia camphorate mainly focuses on the aspects of increasing the quantity of fungi and the yield of polysaccharide, and the amount of fermentation research with the unique active metabolite 4-Acetylantroquinonol B of antrodia camphorate as the target is small. The method starts from strains of antrodia camphorate, by means of optimization of staged fermentation cultivation, cultivation conditions and culture medium composition, 4-Acetylantroquinonol B is obtained from fungi obtained through antrodia camphorate liquid fermentation under the condition that no inductors or precursors are added, and the yield of 4-Acetylantroquinonol B reaches 2.6-5 mg / g. The method is a new method for producing 4-Acetylantroquinonol B through fermentation.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, in particular to a method for fermenting and producing 4-Acetylantroquinonol B (4-Acetylantroquinonol B). Background technique [0002] The fruiting body of Antrodia camphorata parasitizes on the decayed inner wall of the trunk of the camphor tree or lies on the ground near the barkless surface near the soil surface. It has a strong aroma of the camphor tree and contains high content of essential oils. The shape is plate-shaped, bell-shaped, horseshoe-shaped or irregular , perennial, sessile, clinging to the wood surface growth. Many scholars have studied the active components and pharmacological activities of Antrodia camphorata fruiting bodies, and confirmed that the fruiting bodies of Antrodia camphorata contain high-oxidation triterpenoids, active polysaccharides, sterols and other active ingredients. [0003] In recent years, triterpenoids such as 4-AcetylantroquinonolB have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/04C12N1/14C12R1/645
CPCC12N1/14C12P17/04
Inventor 邓利刘欢王萌曾耀铭
Owner 厦门北化生物产业研究院有限公司
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