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Biotransfer process for preparing artemisine compounds

A technology of biotransformation and artemisinin, applied in the direction of fermentation, etc., can solve the problem of no other uses found in the search

Inactive Publication Date: 2002-12-04
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Streptomyces griseus ATCC13273 was used to induce cytochrome P-450 enzymes in three US patents 5,674,714, 5,213,971, and 5,169,755, and no other applications were found

Method used

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  • Biotransfer process for preparing artemisine compounds

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Embodiment 1

[0045] Screening of microbial strains with transforming activity

[0046] In the present invention, 14 strains (strains) are transformed and screened, and the result shows that Streptomyces griseus (ATCC-13273) has strong transformation ability to artemisinin, dihydroartemisinin and artemether.

[0047] Liquid medium for strain screening—soybean flour glucose medium: Take 20g of glucose, 5g of yeast extract, 5g of soybean flour, 5g of NaCl, 5g of K2HPO4, 1000mL of distilled water, and adjust the pH to 7.0 with 6N HCl. Pack in 150ml Erlenmeyer flasks, 25ml per bottle, sterilize at 121°C, 0.15Mpa for 20min.

[0048] When screening strains, the slant solid medium used for preserving strains is to add 1.5% agar to the above-mentioned liquid medium, heat and dissolve, then divide into 15mL test tubes with screw caps, and sterilize at 121°C and 0.15Mpa for 20min. , Obliquely cooled.

[0049] Strain preservation and activation

[0050] The strains were inoculated on the slant soli...

Embodiment 2

[0079] Biotransformation of artemisinin compounds

[0080] Experimental Materials and Instruments

[0081] Microorganisms The microorganisms used in this experiment are Streptomyces griseus, the preservation number of which is ATCC-13273, which has been disclosed in US Patents 5,674,714, 5,213,971, and 5,169,755.

[0082] Reagents and reagents Artemisinin was prepared according to literature methods, see: Chen Yougen, Yu Boyang, etc., Extraction, separation and identification of artemisinin and its precursor compounds, Chinese herbal medicine, 2001; 32(4): 302-303, and identified by spectrum Its structure is consistent with the literature reports; dihydroartemisinin was purchased from Beijing Sixth Pharmaceutical Factory; artemether was purchased from Kunming Pharmaceutical Co., Ltd. The three compounds were respectively dissolved in acetone to make a solution with a concentration of 240 mg / mL, shaken well, and used as conversion substrates for later use.

[0083] Medium 20%...

Embodiment 3

[0109] Selection of Carbon Source, Nitrogen Source and Metal Ion in Artemisinin Microbial Transformation Medium

[0110] The main product of artemisinin transformed by microorganism Streptomyces griseus was screened, especially the transformation conditions of 3α-hydroxyartemisinin.

[0111] Medium:

[0112] Carbon source selection medium: 20% potato decoction 1000ml, carbon source (glucose or soluble starch, sucrose, maltose, citric acid, galactose, dextrin, corn flour, glycerin) 20g, yeast extract 5g, soybean powder 5g , KH 2 PO 4 3.0g, MgSO 4 ·7H 2 O 1.5g, VB 1 Trace amount (pH5.0~7.5).

[0113] Nitrogen source selection medium: 20% potato decoction 1000ml, glucose 20g, nitrogen source (peptone or beef extract, yeast extract, corn flour, soybean flour 10g, or urea, ammonium carbonate, sodium nitrate, L-lysine , DL-α-alanine is equivalent to sodium nitrate 2g / L), KH 2 PO 4 3.0g, MgSO 4 ·7H 2 O 1.5g, VB 1 Trace amount (pH5.0~7.5).

[0114] Metal ion selection me...

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Abstract

A hydroxylation process for biotransferring the artemisine, hydroartemisine, or artemsic methylether, with griseous streptomyces microbe (ATCC 13273) is disclosed. 12 hydroxylated products are separated from the fermented liquid, which include 3 ones for artemisine, 4 ones for dihydroartemisine and 5 ones for artemic methylether, among them, 10 ones are new compounds contg. mother kernels.

Description

technical field [0001] The invention relates to a preparation method for biotransformation of artemisinin compounds, in particular to a method for preparing artemisinin compounds through biotransformation of artemisinin and its derivatives by known microorganisms. Background technique [0002] Microbial transformation is the use of specific enzymes produced by microorganisms to complete specific biochemical reactions. Microorganisms can produce a variety of enzymes that catalyze a variety of reactions. For example, the types of microbial transformation reactions include: oxidation, reduction, hydrolysis, substitution, removal, condensation, and cracking reactions. If a single enzyme can be used to specifically catalyze a specific reaction, by-products can be reduced and the production efficiency of the target product can be improved. Therefore, if you want to use microorganisms to transform and prepare practical target products, you must first screen the strains that produ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18
Inventor 余伯阳陈有根吴志峰
Owner CHINA PHARM UNIV
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