Method for synthesizing betulic acid from betulin through microbial cell bioconversion

A technology of microbial cells and betulin, applied in the fields of bioengineering and microbial fermentation, can solve problems such as research gaps, and achieve the effects of short culture time, high conversion rate and easy operation.

Inactive Publication Date: 2010-11-03
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) Insert Armillaria chrysanthemum ZJUQH spore suspension 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; wherein the potato dextrose liquid and yellow The volume ratio of the Armillaria viridans ZJUQH spore suspension is 30, and the spore concentration in the Armillaria viridans 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) Insert Armillaria chrysanthemum ZJUQH wet cells into the sterilized glucose-containing phosphate buffer (pH is 6, wherein the mass percent concentration of glucose is 2%) to obtain a pre-reaction system; wherein the phosphate buffer The ratio of milliliters to grams of Armillaria chrysanthemum ZJUQH wet cells 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) insert Armillaria chrysanthemum ZJUQH wet cell in the sterilized aqueous glucose solution (mass percentage concentration of glucose is 2%), obtain pre-reaction system; The ratio of wet cells to grams 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 betulin is 0.05g per liter of culture solution;

[0064] (3) The transformed culture fluid is post-treated to obtain betulinic acid.

[00...

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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 reliableconversion 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12R1/645C12P33/00
Inventor 陈启和刘婧章海锋何国庆傅明亮
Owner ZHEJIANG UNIV
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