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Method for preparing demethyl rapamycin by utilizing bacillus megaterium

A technology of methyl rapamycin and Bacillus megaterium, applied in the field of preparation of desmethyl rapamycin, achieving the effects of less environmental pollution, easy acquisition, and no need for group protection

Inactive Publication Date: 2010-10-06
FUJIAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The reports of obtaining desmethylrapamycin by techniques similar to the present invention are extremely limited

Method used

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  • Method for preparing demethyl rapamycin by utilizing bacillus megaterium
  • Method for preparing demethyl rapamycin by utilizing bacillus megaterium
  • Method for preparing demethyl rapamycin by utilizing bacillus megaterium

Examples

Experimental program
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Effect test

example 1

[0040] 1. Insert the Bacillus megaterium preserved in milk into the slant of nutrient agar, culture at 35°C for 15 hours, the slant lawn is plump, pick the platinum ring and insert it into the seed medium (30ml / 250ml triangular flask, g / 1000ml medium) : Soluble starch 24, glucose 1, yeast extract juice powder 3, peptone 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, pH natural, 28 ℃, 220rpm shaking culture after 15h, insert seed culture medium ( 80ml / 500ml Erlenmeyer flask, formula as above), 28°C, 220rpm shaking culture for 8 hours, then transfer the same inoculum amount into the transformation medium (60ml / 500ml Erlenmeyer flask, recipe as above), 28°C, 220rpm shaking culture for 15h, add ethanol to prepare Rapamycin stock solution (15mg / ml), so that the final concentration is 125mg / ml. Then, the transformation culture of rapamycin was carried out at the same temperature and rotation speed, and the transformation was completed after 64 hours, and the transformat...

example 2

[0044] 1. Insert the Bacillus megaterium preserved in milk into the nutrient agar slant, culture at 28°C for 24 hours, the slant lawn is plump, pick the platinum ring and insert it into the seed medium (30ml / 250ml triangular flask, g / 1000ml medium) : Corn starch 30, sucrose 10, soybean powder 10, peptone 2, sodium chloride 1, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, natural pH, 28°C, 220rpm shaking culture for 24 hours, inoculation transformation with 3% inoculum Culture medium (60ml / 500ml Erlenmeyer flask, formula as above), 28°C, 220rpm shaking culture for 18h, add rapamycin stock solution (15mg / ml) prepared with ethanol to make the final concentration 125mg / ml. Then, the transformation culture of rapamycin was carried out at the same temperature and rotation speed, and the transformation was completed after 72 hours, and the transformation liquid was collected.

[0045] 2. Take the transformation liquid, and extract twice with the ratio of transformation liq...

example 3

[0047] 1. Insert the Bacillus megaterium preserved in milk into the nutrient agar slant, culture at 32°C for 18 hours, the slant lawn is plump, pick-platinum ring into the seed medium (30ml / 250ml triangular flask, g / L) dextrin 30 , fructose 5, peanut cake powder 10, peptone 1, yeast extract powder 3, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, pH natural, 28 ℃, 220rpm shaking culture 24h, insert transformation medium with 2% inoculum size ( 60ml / 500ml Erlenmeyer flask, formula as above), 28°C, 220rpm shaking culture for 24h, add rapamycin mother solution (15mg / ml) prepared with ethanol, so that the final concentration is 125mg / ml. Then, the transformation culture of rapamycin was carried out at the same temperature and rotation speed, and the transformation was completed after 72 hours, and the transformation liquid was collected.

[0048] 2. Take the transformation liquid, and extract twice with the ratio of transformation liquid: ethyl acetate (1:1), 20 minutes ...

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Abstract

The invention relates to a method for preparing demethyl rapamycin by utilizing bacillus megaterium, including that bacillus megaterium is inoculated to slope culture medium to be cultured and then is inoculated into seed culture medium to be cultured, and then the seed culture solution is inoculated into a transformation culture medium. When the bacillus megaterium propagates in the transformation culture medium, substrate rapamycin is added, the bacillus megaterium carries out transformation on the rapamycin, so as to obtain bacillus megaterium transformed rapamycin transformation solution, and the transformation solution contains 29, 42-O-didemethyl rapamycin, 42-O-demethyl rapamycin and 29-O-demethyl rapamycin and other rapamycin derivates. The invention has the advantage that micro organism is adopted to obtain rapamycin derivate. Pollution to the environment is less, action condition is mild, and no group protection is required, and transformation efficiency of rapamycin derivate can be improved by improving micro organism and condition thereof for transforming rapamycin.

Description

technical field [0001] The invention relates to a preparation method of desmethylrapamycin. Background technique [0002] Rapamycin (Rapamycin; Sirolimus, Sirolimus) is a macrolide compound produced by the fermentation of Streptomyces hygroscopicus AY B-994 and Streptomyces hygroscopicus FC904 with immunosuppressive, antifungal, antiproliferative and antitumor activities ( figure 1 ), a mammalian target of rapamycin C1 (mTORC1) inhibitor. It has been used clinically as an anti-rejection drug for kidney transplantation, and a rapamycin-coated stent for the prevention and treatment of coronary artery restenosis after stent implantation; it is being used as an anti-tumor drug (anti-leukemia, breast cancer, gastric cancer) that inhibits the mTOR signal transduction pathway , intestinal cancer, liver cancer, etc.) for research or clinical trials; the latest research shows that rapamycin has the effect of prolonging the life cycle of old mice. [0003] The chemical semi-synthesi...

Claims

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Application Information

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IPC IPC(8): C12P17/14C07D498/18C12R1/11
Inventor 黄捷程元荣杨国新金东伟余辉李夸良陈夏琴陈有钟
Owner FUJIAN INST OF MICROBIOLOGY
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