Method for preparing bilobalide B by using Stereum hirsutum thalli as catalyst

A technology of ginkgolide and tremella fungus, which is applied in the field of bioengineering and can solve problems such as difficulty in mass production of ginkgolide and the like

Active Publication Date: 2013-12-25
BEIJING LYUKE TIANCHENG BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And there are only nine articles about the preparation of ginkgolide B (a new method for extracting ginkgolide B from ginkgo biloba or ginkgo leaf extract, application number 200510063407.X; a kind of separation of ginkgolide from ginkgolide mixture The method of B, application number: CN201010608847.X, etc.), the ginkgolides contain ginkgolides A, B, C, M and other homologues in the ginkgolides, these patents adopt separation technology to separate ginkgolides from the ginkgolide mixture B, it is difficult to produce ginkgolides in large quantities in industry B

Method used

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  • Method for preparing bilobalide B by using Stereum hirsutum thalli as catalyst

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1 Preparation of test tube strains

[0015] Preparation of potato dextrose agar medium: Weigh 200 g of potatoes, cut into pieces, add 1000 mL of water, boil for 30 minutes, filter, dilute the filtrate to 1000 mL, add 20 g of agar, heat until the agar is completely melted, and then divide into 18 × 200 mm 15mL of culture medium per tube, sterilized at 120°C for 30 minutes, cooled to 50°C, placed on a slope, the length of the slope is half the length of the test tube; the preserved Tremella strains isolated from the fruiting bodies of Tremella were cut into 4× 4mm small pieces of strain, pick a small piece and transfer it to a test tube containing potato dextrose agar medium, and cultivate at 20°C for 20 days to obtain a test tube slanted strain, which is stored at 4°C for future use:

Embodiment 2

[0016] Example 2 Preparation of the fermentation broth containing ginkgolide B

[0017] 1. Preparation of liquid shake flask strains

[0018] The formula of liquid shake flask culture medium is (unit is g / L) glucose 5, corn meal 5, peptone 1, KH 2 PO 4 1. MgSO 4 1. The initial pH is 5.0. A total of 5500mL was prepared, divided into 250mL conical flasks, each 60mL, a total of 84 bottles, sterilized at 100 ℃ for 60 minutes. A total of 5500mL was prepared, and 250mL conical flasks were divided into 250mL conical flasks, each 60mL, a total of 84 flasks. The preserved Tremella test tube slanted strains isolated from the fruiting bodies of Yunnan Tremella were cut into small pieces of 4×4mm size, and 3 pieces were picked for transfer. Put it into a 250 mL conical flask containing a shake flask medium, and place it in a shaker at 22°C at 90 rpm for 24 hours.

[0019] 2. Preparation of seed pot strains

[0020] The 75L seed tank contains 45L seed medium, wherein the formula of t...

Embodiment 3

[0025] Example 3 Preparation of the fermentation broth containing ginkgolide B

[0026] 1. Production of shake flask strains

[0027] The formula of the shake flask medium is (in g / L): glucose 20, corn meal 15, peptone 6, KH 2 PO 4 3. MgSO 4 2. The initial pH value is 6.5, a total of 4500mL is prepared, and 250mL conical flasks (50 bottles in total) are divided into 90mL bottles, and sterilized at 120°C for 40 minutes. After cooling, the preserved Tremella test-tube slanted strains separated from the fruiting bodies of Shanxi Tremella were cut into small pieces of 4×4 mm size, and the test-tube slanted strains were cut into small pieces of 4×4 mm size, and 4 pieces were picked and transferred to Placed in a conical flask with shake flask medium, placed on a shaker at 25°C, 120 rpm, and cultured for 36 hours.

[0028] 2. Preparation of seed pot strains

[0029] The 100L seed tank contains 50L seed medium, and the formula of the seed tank medium is (in g / L): glucose 20, cor...

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Abstract

The invention discloses a method for synthesizing bilobalide B by using Stereum hirsutum thalli as a catalyst to convert a bilobalide mixture, belonging to the biotechnical field. The method disclosed by the invention comprises the following steps: carrying out enlargement cultivation on the Stereum hirsutum thalli; converting bilobalide to synthesize the bilobalide B by using the thalli as a catalyst by means of a test tube slant, a liquid shake flask, seed culture and thallus separation; and centrifugalizing the conversion liquid, extracting by ethyl acetate and crystallizing by anhydrous alcohol to separate the bilobalide B from fermentation liquor. The purity of the bilobalide B obtained by the method is greater than or equal to 99%, and the bilobalide conversion ratio is greater than 90%.

Description

technical field [0001] The invention relates to the technical field of bioengineering, and relates to a new process for preparing ginkgolide B through liquid culture of Tremella fungus bacteria and simultaneous transformation of Ginkgo biloba extract. Background technique [0002] Studies have shown that ginkgolide is one of the main active components in Ginkgo biloba, and is a kind of specific and effective platelet activating factor (PAF) receptor antagonist. Platelet activating factor is an endogenous phospholipid secreted by platelets and various inflammatory tissues. It is the most effective platelet aggregation inducer found so far, and is closely related to the occurrence and development of many diseases. Ginkgolide, as a specific antagonist of PAF receptors, has a wide range of pharmacological effects. Studies have found that ginkgolide B has protective effects on the central nervous system and ischemic injury, anti-shock, anti-allergic, antibacterial and anti-infla...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12R1/645
Inventor 张志才
Owner BEIJING LYUKE TIANCHENG BIO TECH
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