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Method for producing Vinca rosea alkaloid

A periwinkle and culture medium technology, applied to plant cells, etc., can solve the problems of chemical synthesis method production, practical application limitations, and high market prices, and achieve low production costs, industrialized production, and short cultivation cycles. Effect

Inactive Publication Date: 2010-06-02
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the content of these two alkaloids in plants is very low, respectively a few parts per hundred thousand and a few parts per million, and cannot be produced by chemical synthesis, the output of vinblastine and vincristine is far from satisfying the market demand, leading to high market prices, practical applications are greatly restricted

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, utilizing periwinkle in vitro cell large-scale culture to produce periwinkle alkaloids

[0026] 1. Obtaining periwinkle callus

[0027] Use the young stems of periwinkle as explants, sterilize the surface with 70% ethanol for 30-50 seconds, put them into 5-15% sodium hypochlorite solution for secondary sterilization for 10-20 minutes, and then rinse them with sterile water three times. Cut the young stems into 2 mm long stems in the ultra-clean workbench, inoculate them into the dedifferentiation medium, and cultivate them under 2000 lx, 12-16 hours of light, and room temperature of about 25 degrees for 3-4 weeks to obtain white cured stems. damage tissue. Then the callus is placed in the subculture acclimation medium described in step 2 and subcultured for more than 3 times, and at the same time, a cell line with fast growth rate, soft texture and stable growth amount is obtained through screening.

[0028] Preparation of dedifferentiation medium: in MS...

Embodiment 2

[0040] Embodiment 2, utilizing periwinkle in vitro cell large-scale culture to produce periwinkle alkaloids

[0041] 1. Obtaining periwinkle callus

[0042] Use the internodes of periwinkle as explants, sterilize the surface with 70% ethanol for 30-50 seconds, put them into 5-15% sodium hypochlorite solution for secondary sterilization for 10-20 minutes, and then rinse them with sterile water three times. Cut the internodes into 2 mm long stems and inoculate them into the dedifferentiation medium in an ultra-clean workbench, and culture them at 2000 lx, 12-16 hours of light, and room temperature of about 25 degrees for 3-4 weeks to obtain white calluses organize. Then the callus is placed in the subculture acclimation medium described in step 2 and subcultured for more than 3 times, and at the same time, a cell line with fast growth rate, soft texture and stable growth amount is obtained through screening.

[0043] Preparation of dedifferentiation medium: in MS medium, add i...

Embodiment 3

[0055] Embodiment 3, utilizing periwinkle in vitro cell large-scale culture to produce periwinkle alkaloids

[0056] 1. Obtaining periwinkle callus

[0057] Use the leaves of periwinkle as explants, sterilize the surface with 70% ethanol for 30-50 seconds, put them into 5-15% sodium hypochlorite solution for secondary sterilization for 10-20 minutes, then rinse them with sterile water three times, take them out and put them in In the ultra-clean workbench, cut the leaf veins into small pieces of 0.5 cm square and inoculate them in the dedifferentiation medium, and cultivate them at 2000 lx, 12-16 hours of light, and at room temperature of about 25 degrees for 3-4 weeks, and then white callus can be obtained. organize. Then the callus is placed in the subculture acclimation medium described in step 2 and subcultured for more than 3 times, and at the same time, a cell line with fast growth rate, soft texture and stable growth amount is obtained through screening.

[0058] Prep...

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PUM

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Abstract

The invention discloses a method for producing catharanthus roseus alkaloid. The method comprises the following steps: firstly, dedifferentiation culture is performed on the explant of the catharanthus roseus to get the callus of the catharanthus roseus, and through succesive transfer culture, loose and quick-growing cell line is obtained; secondly, the cell line of the callus of the catharanthusroseus obtained in the step one is processed through inducing the culture in basal culture medium provided with colchicines to get polyploidy cell line, and then expanding culture is performed to geta large amount of catharanthus roseus polyploidy cells; thirdly, the catharanthus roseus polyploidy cells are inoculated into a deffined medium to get cells being rich ofcatharanthus roseus secondarymetabolites. The method of the invention produces the catharanthine with the catharanthus roseus, the industrial production of the catharanthine can be firmly realized, the production cost is low, theculture period is short, the production is not influenced by natural environment and climate, and the year-round production can be realized.

Description

technical field [0001] The invention relates to a method for producing vinca alkaloids. Background technique [0002] Periwinkle (Catharanthus roseus (L) G. Don) is a genus with rich alkaloid content in Apocynaceae. Vinca alkaloids mainly include: Aspicospermatan type such as Vindoline (Vindoline) etc., Ibogan type such as Vinblastine (Catharanthine) etc., Corynanthean type such as Ajmalicine (Ajmalicine) and Serpentine (Serpentine) etc. and Bisindoles type such as Vinblastine and Vincristine etc. [0003] In the mid-1950s, some scholars found that the extract of periwinkle plants also had the effect of lowering white blood cells when studying the hypoglycemic effect of periwinkle plants. In 1958, Canadian Noble and Beer jointly isolated this active ingredient, named it Vinblastine (vinblastine), and prepared its sulfate. [0004] Subsequently, Lily Company of the United States proved that vinca alkaloids can treat acute lymphoblastic leukemia; Vinblastine is mainly used...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04C07D519/04
Inventor 郭志刚刘云
Owner TSINGHUA UNIV
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