Method for synthesizing salidroside and icarisid D2 through biotransformation and application thereof
A technology for icariside and salidroside, which is applied in the field of biotransformation and synthesis of salidroside and icariside D2, can solve the problems of many by-products, high loss of raw materials, high cost, etc., and achieve the required Short time, high conversion rate, and the effect of reducing loss
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Embodiment 1
[0022] 1. Leonurus suspension cell culture: Take 5g of the purified and vigorously growing Leonurus loose callus, insert it into 100mL culture medium (MS medium+0.5mg / L 2,4-dichlorophenoxyacetic acid+2.0mg / L6- benzylaminoadenine) in a 250 mL shake flask. According to the new solution: old solution = 3:1, change the solution every 5 days. In the early stage of culture, remove large cell clusters each time the medium is changed. After cultivating for a period of time, when there are sufficient small cell clusters, pass the suspended cell liquid through a 50-mesh sieve when changing the medium, harvest the filtrate, let it stand for a while until the cells sink, and discard the upper part with a ratio of new solution to old solution of 3:1. solution, supplemented with new solution, and continued the shaker culture. After repeatedly passing through a 50-mesh sieve for a period of time, the motherwort suspension cell line was established. Thereafter, pass through a 50-mesh sieve...
Embodiment 2
[0029] 1. Motherwort suspension cell culture: the culture method is the same as in Example 1.
[0030]Transformation method: add an aqueous solution of p-hydroxyphenylethanol to the transformation system of motherwort suspension cells to make the initial concentration 5mmol / L, and harvest 24h and 48h after the addition, and harvest twice in total.
[0031] A blank group is set at the same time, the difference between the blank group and the present embodiment is only that the blank group does not add p-hydroxyphenylethanol aqueous solution.
[0032] 2. Salidroside and Icariside D 2 The detection of: detection method is with embodiment 1.
[0033] It can be seen from the detection results that the total conversion rate of salidroside in the Leonurus suspension cell transformation system obtained in this example (29.78% in the cell + 5.71% in the culture medium) in 24 hours was 35.49%, and icariside D 2 The total conversion rate (5.03% in cells + 1.02% in culture medium) was 6...
Embodiment 3
[0035] 1. Motherwort suspension cell culture: the culture method is the same as in Example 1.
[0036] Transformation method: add an aqueous solution of p-hydroxyphenylethanol to the transformation system of motherwort suspension cells to make the initial concentration 7mmol / L, harvest 1h and 48h after the addition, and harvest twice in total.
[0037] A blank group is set at the same time, the difference between the blank group and the present embodiment is only that the blank group does not add p-hydroxyphenylethanol aqueous solution.
[0038] 2. Salidroside and Icariside D 2 The detection of: detection method is with embodiment 1.
[0039] It can be seen from the detection results that the total conversion rate of salidroside in the Leonurus suspension cell transformation system obtained in this example (3.40% in the cell + 1.09% in the culture medium) in 1h was 4.49%, and icariside D 2 The total conversion rate (1.08% in cells + 0.76% in culture medium) was 1.84%; the to...
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