Method for preparing ginsenoside Rd by utilizing microbial conversion

A microbial transformation and ginsenoside technology, applied in the field of biomedicine, can solve the problems of single acquisition channel, low content, complex structure, etc., and achieve the effect of expanding the source of drug resources, simple process, and high yield

Inactive Publication Date: 2013-01-23
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

At present, domestic Guangdong Taihe Pharmaceutical Technology Co., Ltd. has developed Rd injection as a new drug for the treatment of cerebral apoplexy, and has entered the phase III clinical trial summary stage, but the medicinal source of Rd is mainly directly extracted from natural ginseng saponins; For well-known reasons, because the natural Rd content is very rare and the acquisition channel is single, its market price is very expensive
[0004] It is well known in the art that the content of ginsenoside Rd in ginseng is relatively low, only about 0.2%. Because of its complex structure, the chemical synthesis of this ginsenoside Rd has not been successful so far; The Rd monomer is obtained by extracting from the leaves, because the extraction method yield is low and the cost is high, which obviously affects the further expansion of the medical care market demand, so it is necessary to develop a kind of high yield, good purity, and simple and easy preparation The method of taking ginsenoside Rd
At present, there are studies trying to convert the main saponins into rare saponins with higher activity by adding chemical methods, but the hydrolysis conditions in the above-mentioned chemical methods are severe and difficult to control, and the saponins often undergo dehydration, cyclization, and differentiation during the hydrolysis process. Reactions such as isomerization and hydroxylation produce many by-products, and it is difficult to obtain the target product; however, due to the strong regioselectivity and mild reaction conditions of biotransformation, researchers in this field favor enzymatic transformation and microbial transformation Rare ginsenosides are prepared by the method; but there are still defects and deficiencies as follows in the obtained ginsenoside Rd and its conversion process:
Ye L. et al obtained a highly transformed strain Paecilomyces Bainier sp.229-7 by LiCl mutagenesis, and carried out 6L fermentation, the conversion rate can reach 90%, but the purity of the final product obtained is only 92%

Method used

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  • Method for preparing ginsenoside Rd by utilizing microbial conversion
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  • Method for preparing ginsenoside Rd by utilizing microbial conversion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 strain screening

[0045] Purchase 50 strains of different strains from the Industrial Microbiology Collection Center of China Microbiological Culture Collection Management Committee, and use the following shake flask process to screen:

[0046] Fermentation medium: 3% sucrose, 3% soybean flour, 0.1% ammonium sulfate, 0.1% magnesium sulfate, 0.1% calcium chloride, 0.5% bran. All culture media were packed in 250ml Erlenmeyer flasks with a volume of 30ml, and sterilized at 121°C for 20min. Inoculate from potato slant, culture on shaker at 28°C and 250rpm for 24h, add 5% ginsenoside Rb1, and continue to cultivate for 48h. After the fermentation is over, an equal volume of ethanol is added for extraction. Through HPLC analysis, it is determined that Aspergillus niger with strain preservation number CICC40426 is the bacterial strain for preparing ginsenoside Rd through microbial transformation of the present invention, and it has the best conversion rate, which c...

example 2

[0047] Example 2 passage test

[0048] Using the fermentation method of Example 1, ferment the Aspergillus niger whose strain preservation number is CICC40426, and carry out the passage test. The results are shown in Table 1. After 20 generations of passage, the Rd conversion rate of the strain is still very stable.

[0049] Table 1 Passage results

[0050]

[0051]

example 3

[0052] The optimization of example 3 culture medium

[0053] Using the fermentation method of Example 1, ferment the Aspergillus niger with the strain preservation number CICC40426, optimize the medium, and optimize the carbon source: glucose, sucrose, maltose, lactose, potato, starch, and bran were respectively investigated as carbon sources. , the influence on the conversion rate of Rd. Results When bran, glucose, sucrose and starch were used as carbon sources, the conversion rate of Rd was higher. Delayed nitrogen source: The effects of corn steep liquor, cottonseed powder, yeast powder, soybean powder, peptone and peanut powder on the conversion rate of Rd were investigated respectively as delayed nitrogen sources. Results When yeast powder, soybean powder and peptone were used as nitrogen sources, the conversion rate of Rd was higher. On this basis, statistical methods are used to comprehensively investigate the effects of carbon sources, nitrogen sources, trace element...

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Abstract

The invention belongs to the technical field of biological medicine, relates to a method for preparing ginsenoside Rd by utilizing microbial conversion and in particular relates to a microbial conversion method for preparing the ginsenoside Rd by fermenting and converting aspergillus niger into various glycol-type ginsenosides. In the method disclosed by the invention, a high-conversion industrial aspergillus niger strain with culture preservation number being CICC40426 is adopted, various glycol-type ginsenosides are obtained by fermentation and conversion in a full-automatic fermentation tank, and high-purity ginsenoside Rd is prepared through purification and crystallization by virtue of macroporous resin. The method disclosed by the invention is simple in process, low in cost and high in yield and is environment-friendly; and with adoption of the method disclosed by the invention, industrial preparation of the ginsenoside Rd can be realized, source of the ginsenoside Rd drug resource is expanded, and wide medical care market demand is met, so that the method disclosed by the invention has great value.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to a method for preparing ginsenoside Rd by microbial transformation, in particular to a microbial transformation method for preparing ginsenoside Rd by fermenting and transforming various diol-type ginsenosides with Aspergillus niger. technical background [0002] Ginseng (Panax ginseng C.A. Meyer) is a traditional Chinese medicinal material with complex chemical components, extensive biological activities and unique pharmacological effects. With the advancement of modern separation and analysis techniques, the chemical components in ginseng have been further elucidated. Ginsenosides are the main biologically active substances in these isolated components. So far, there are 50 ginsenoside monomers that have been isolated and identified. More than one species. [0003] Studies have disclosed that ginsenoside Rd has been confirmed by Guan Yongyuan, School of Medicine, Sun Yat-Sen U...

Claims

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Application Information

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IPC IPC(8): C12P33/20C12R1/685
Inventor 周珮史训龙周伟冯美卿李继扬叶丽周超群
Owner FUDAN UNIV
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