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Method for promoting biosynthesis of 10-hydroxycamptothecine

A technology of hydroxycamptothecin and biosynthesis, which is applied in the biological field to achieve the effects of low production cost, short production cycle and favorable protection

Inactive Publication Date: 2011-07-20
赵荣国
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Problems solved by technology

At present, the research on the preparation of 10-hydroxycamptothecin includes chemical total synthesis, chemical semi-synthesis, biotransformation and searching for new drug sources in related plants, etc., but due to various reasons, these methods have not yet been used in industrialized large-scale production

Method used

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Embodiment Construction

[0036] The present invention will be further described below in conjunction with embodiment, can make those skilled in the art understand the present invention more comprehensively, but do not limit the protection scope of the present invention in any way.

[0037] 1. Establishment of Campylobacteria cell culture line:

[0038] Induction and subculture of A, camptophyllin callus:

[0039] Preparation of medium for inducing callus: prepare one liter of MS medium with distilled water. Add 0.1mg / L 6-benzyladenine, 30g / L sucrose, 2g / L urea, 0.25mg / L 2,4-dichlorophenoxyacetic acid, the coagulant is agar 6g / L, and the pH is 5.8. Divide into 25 100mL Erlenmeyer flasks evenly, place in a pressure cooker after stoppering, and sterilize for 30 minutes at 121°C and 15 atmospheres.

[0040] Select the leaves of Campylophora radicis as explants, first clean them with clean water, soak them in 70% ethanol and rinse them for 5 minutes, take them out and rinse them with clean water three ti...

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Abstract

The invention belongs to technical filed of biology, in particular to a method for promoting biosynthesis of 10-hydroxycamptothecine. The method comprises the following steps of: constructing a suspension culture system of camptotheca acuminata cells for an explant by using leaves of camptotheca acuminata plants; adding three kinds of exogenous substances such as cobalt nitrate, lanthanum nitrate and salicylic acid to regulate and control the secondary metabolism of the camptotheca acuminata cells; and inoculating two kinds of microorganisms such as Mucor spinosus and Aspergillus niger for biotransformation at the later stage of cell culture, placing the treated camptotheca acuminata cell culture into a pulp refiner, into which an ethanol solution is added, for homogenating, and filtering to obtain filtrate containing 10-hydroxycamptothecine. The method has the characteristics of short production period, low production cost and high yield of 10-hydroxycamptothecine, and is beneficial to the protection of the camptotheca acuminata plant resources.

Description

technical field [0001] The invention belongs to the field of biotechnology. It specifically relates to a method for promoting the biosynthesis of 10-hydroxycamptothecin. Background technique [0002] 10-Hydroxycamptothecin is a trace alkaloid contained in Camptotheca acuminala Decne, a unique plant in my country. It targets DNA topoisomerase I and inhibits the synthesis of DNA in organisms. And play an anti-tumor effect. It is not only an antitumor drug with better clinical effect itself, but also an important intermediate for preparing other camptothecin derivatives such as Topotecan and Irinotecan; it has clinically shown It has a wide range of anti-tumor activities and relatively small toxic and side effects. The domestic clinical demand and export volume are increasing year by year. For a long time, my country's enterprises have been directly extracting from camphor tree plants, but because of its low content (about 0.002%) and high production costs, it is difficult to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P17/18C12R1/685C12R1/785
Inventor 赵荣国
Owner 赵荣国
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