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.
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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.
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