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Method for synthesizing betulic acid from betulin through microbial cell bioconversion

A technology of microbial cells and betulin, which is applied in the fields of bioengineering and microbial fermentation, can solve problems such as research blanks, and achieve the effects of short cultivation time, high conversion rate, and safe and reliable conversion process

Inactive Publication Date: 2009-06-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There are very few studies on the biological synthesis of betulinic acid. There are only some researches on the microbial transformation of betulinic acid and its derivatives in foreign countries, while domestic research is almost blank.

Method used

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  • Method for synthesizing betulic acid from betulin through microbial cell bioconversion
  • Method for synthesizing betulic acid from betulin through microbial cell bioconversion
  • Method for synthesizing betulic acid from betulin through microbial cell bioconversion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Synthesis of betulinic acid by biotransformation of betulin by direct growth transformation method

[0044] (1) Add the spore suspension of Clostridium chrysogenum ZJUQH into the sterilized potato dextrose liquid, and cultivate it in a shaker at 28°C with a rotation speed of 120r / min for 3 days to obtain a pre-reaction system; The volume ratio of the ZJUQH spore suspension is 30, and the spore concentration in the Armillaria chrysanthemum ZJUQH spore suspension is 1×10 6 a / ml;

[0045] (2) Add betulin solution (wherein the concentration of betulin is 7.5 mg / ml) to the pre-reaction system obtained in step (1), and transform in a shaker at 28° C. with a rotating speed of 120 r / min for 6 days to obtain transformation After the culture fluid, betulin consumption is calculated as 0.05g per liter of potato glucose liquid;

[0046] (3) The transformed culture fluid is post-treated to obtain betulinic acid.

Embodiment 2

[0057] Example 2 Synthesis of betulinic acid by biotransformation of betulin by quiescent cell transformation method

[0058] (1) Add the wet cells of Clostridium chrysogenum ZJUQH into the sterilized glucose-containing phosphate buffer (pH is 6, wherein the mass percent concentration of glucose is 2%) to obtain the pre-reaction system; wherein the milliliter of the phosphate buffer The ratio of the number to the gram number of the wet cells of Closesia chrysogenum ZJUQH is 5;

[0059] (2) Add betulin solution (wherein the concentration of betulin is 7.5 mg / ml) to the pre-reaction system obtained in step (1), and convert and react in a shaker at 28° C. with a rotating speed of 120 r / min for 6 days to obtain For the culture fluid after conversion, the consumption of betulin is 0.05g per liter of culture fluid;

[0060] (3) The transformed culture fluid is post-treated to obtain betulinic acid.

Embodiment 3

[0061] Example 3 Synthesis of betulinic acid through biotransformation of betulin by microemulsion system transformation method

[0062] (1) Into the sterilized aqueous glucose solution (the mass percent concentration of glucose is 2%), insert the wet cells of Clostridium chrysogenum ZJUQH to obtain the pre-reaction system; wherein the milliliters of aqueous glucose solution is the same as the gram of the wet cells of Clostridium chrysogenum ZJUQH The ratio of the numbers is 5;

[0063] (2) Add the betulin solution containing absolute ethanol and Tween 80 in the pre-reaction system that step (1) obtains (by volume ratio, absolute alcohol: Tween 80: dimethyl methylene of betulin Sulfone solution=8:1:2.25, the concentration of betulin in the dimethyl sulfoxide solution of betulin is 7.5 mg / ml), and the transformation reaction was carried out in a shaker at 28° C. with a rotation speed of 120 r / min for 6 days to obtain the transformed For the culture solution, the dosage of betu...

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Abstract

The invention discloses a method for synthesizing betulic acid by microbial cell bioconversion of betulin comprising the following steps: inoculating an Armillaria luteo-virens Sacc. ZJUQH to sterilized culture solution and obtaining a pre reaction system; adding a substrate solution containing the betulin and reacting for 5-7 days at between 25 and 30 DEG C, processing the converted culture solution and obtaining the betulic acid, wherein the betulin dosage is 0. 01-0. 1 g per each litre of culture solution. Comparing with the direct plant extraction of the betulic acid method and the chemical synthesis of the betulic acid method The method of the invention has advantages of short microbial cell culture time, easy operation and control, short whole bioconversion period, safe and reliable conversion process, low cost and is suitable for the industrial production.

Description

technical field [0001] The invention relates to the fields of bioengineering and microbial fermentation, in particular to a method for synthesizing betulinic acid through microbial conversion reaction. Background technique [0002] Betulin (betulin, lup-20(29)-ene-3β, 28-diol) and betulinic acid (betulinic acid, 3β-hydroxy-lup-20(29)-en-28-oic acid) belong to lupane type Triterpenoids exist widely in nature, and the most abundant natural resource is birch (Betula spp.). In recent years, scientific research has found that betulin and betulinic acid are very valuable natural products. Betulin, betulinic acid and their derivatives have shown great potential as biological agents in anti-AIDS (HIV) and cancer treatment, and have shown a different mechanism of action from previous drugs. Therefore, the research on betulin, betulinic acid and their derivatives has become a hot spot in the research of natural organic medicines in recent years. Betulinic acid, in particular, has a...

Claims

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Application Information

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IPC IPC(8): C12P33/00C12R1/645
Inventor 陈启和刘婧章海锋何国庆傅明亮
Owner ZHEJIANG UNIV
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