Process for preparing rare ginsenoside Compound K by fermenting panax notoginseng saponins with streptomycete

A technology of notoginseng saponins and ginsenosides, applied in the fields of medicine and biology

Inactive Publication Date: 2008-03-12
KUNMING NOVOGINSENG BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are no reports in the literature at home and abroad on the research involving Streptomyces fermentative...

Method used

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  • Process for preparing rare ginsenoside Compound K by fermenting panax notoginseng saponins with streptomycete
  • Process for preparing rare ginsenoside Compound K by fermenting panax notoginseng saponins with streptomycete

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1 Isolation of bacterial strains: cut the fresh notoginseng stems collected from Wenshan County, Yunnan Province into small sections (3-4cm), soak them in 70% alcohol for 0.5min, carry out surface disinfection, and then sterilize with 0.1% mercuric chloride Take it out for 8 minutes, wash it with sterile water 4 to 5 times, inoculate it on MS medium of plant tissue, firstly white granular hyphae with a diameter of 0.1-0.5mm will appear around the incision of the stem of Panax notoginseng, and gray powder will appear on the mycelia after 6 days spores and cover the stem cut. Pick the gray spores from the stem and place them on the escin-iron citrate plate medium, and after culturing at 28°C for 12 hours, irregular light brown-green hydrolysis circles appear, indicating that the strain has extracellular β-glucosidase activity. Inoculate the gray spores at the stems into the solid medium of Panax notoginseng powder for cultivation, and continue to transfer after a ...

Embodiment 2

[0020] Example 2 Streptomyces fradiae NTGA-334 was transferred from Gaoshi No. 1 slant to ISP2 seed medium containing 1% (w / v) Panax notoginseng saponins, cultured at 28°C for 12h; then inoculated Into 60L humic acid culture medium containing 3000g Panax notoginseng saponins, and ferment at 28°C in a 100L fully automatic fermenter. After 36 hours, adjust and increase the ventilation ratio to 1:3 (v / v), and adjust the stirring rate to 400r / min. After 48 hours, adjust the fermentation temperature to 40°C, and dynamically adjust the pH to 5.0 with 5% (w / v) ammonia water. After 72 hours of fermentation, add Amberlite XAD-16 macroporous adsorption resin at 3% (w / v), stir for 2 hours, and put Can. The fermented liquid was filtered through a filter cloth to obtain a precipitate with a wet weight of 5112 g. The precipitate was ultrasonically extracted with 30 L of ethanol (95%) for 2 hours and filtered through a filter cloth. The filter residue was washed 5 times with 1 L of ethanol ...

Embodiment 3

[0021] Example 3 Streptomyces fradiae NTGA-334 was transferred from Gaoshi No. 1 slant to ISP2 medium containing 1% (w / v) Panax notoginseng saponins, cultured at 28°C for 12h; then inoculated In the 60L humic acid medium containing 3000g of Panax notoginseng saponins, ferment at 28°C in a 100L fully automatic fermenter. After 36 hours, adjust and increase the ventilation ratio to 1:2 (v / v), and adjust the stirring rate to 350r / min. After 48 hours, adjust the fermentation temperature to 38°C, and dynamically adjust the pH to 5.0 with 5% (w / v) ammonia water. After 96 hours of fermentation, add DH101 macroporous adsorption resin at 4% (w / v), stir for 4 hours, and put it into the tank. The fermented liquid was filtered through a filter cloth to obtain a precipitate with a wet weight of 5909 g. The precipitate was ultrasonically extracted with 20 L of ethanol (95%) for 3 h and filtered through a filter cloth. The filter residue was washed 5 times with 1 L of ethanol (95%). The eth...

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Abstract

The invention pertains to the technical field of biology and medicine, and relates to a bacterial strain of streptomyces fradiae NTGA-334 with a preseravation no. of CGMCC No.2074, and a process for using the bacterial strain to ferment and transform Compound K in glycol group in various notoginsenosides so as to prepare 20-O-beta-D-glucopyranoside-20 (S)-protopanaxadiol. The culture media are Gause's no.1, ISP2, corroding acid (HSG culture medium) and asparagine culture medium. During the fermentation, the dosing proportions of various notoginsenosides are 0.1-10% (w/v), fermenting temperature is 18-50 DEG C, dynamically adjusted pH value of 0.1-35%(w/v) aqueous ammonia is 0.1-30% (w/v). The invention transforms and ferments in a scaled way various notoginsenosides, prepares Compound K, provides a streptomycete stain that is of steady condition, simple nutrition requirement and good growth, and provides a process for fermenting and separating by a simple and easy fully automatic fermenting tank. Using this bacterial strain and process to produce Compound K, the production cost is low, the foreign matter is less, and the yield rate is high.

Description

technical field [0001] The invention belongs to the technical fields of biology and medicine, and relates to a kind of Streptomyces fermenting and transforming diol group ginsenosides in various notoginseng saponins, and preparing 20-O-β-D-glucopyranosyl-20(S)-protoginseng Diol (ginsenoside Compound K, hereinafter referred to as Compound K) technology. In particular, it provides an artificially domesticated Streptomyces fradiae NTGA-334 strain, and a process for fermenting and transforming various notoginseng saponins and separating and extracting Compound K by using the strain in an automatic fermenter. Background technique [0002] Rb 1 , Rb 2 , Rc, Rg 3 and Rh 2 Various natural diol group ginsenosides are reported to have antitumor effect (SteveHelms ND: Alternat.Med.Rev., 2004, 9 (3), 259-274; Shoji S: J.Korean Med.Sci., 2001, 16 (Suppl.), S28-37.). Human metabolism experiments show that ginsenosides after oral administration only undergo slight oxidation under the...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P33/00C12R1/54
Inventor 文孟良李铭刚杜刚赵江源
Owner KUNMING NOVOGINSENG BIOENG
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