Microbial transformation preparation method for hydroxycamptothecin

A technology for microbial transformation and hydroxycamptothecin, applied in the biological field, can solve the problems of unutilized large-scale production, difficult to meet clinical needs, and high production costs, and achieve the effects of fewer separation steps, saving time and solvent, and simple separation methods.

Inactive Publication Date: 2009-08-19
DALIAN UNIV OF TECH
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Problems solved by technology

Hydroxycamptothecin is an alkaloid in the plant camptotheca. For a long time, Chinese enterprises have been directly extracting it from plants, but because of its low content and high production costs, it is difficult to meet clinical needs.
Subsequently, people have carried out a lot of

Method used

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  • Microbial transformation preparation method for hydroxycamptothecin
  • Microbial transformation preparation method for hydroxycamptothecin

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Embodiment Construction

[0033] 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.

[0034] 1. Establishment of Campylobacteria cell culture system:

[0035] Induction and subculture of A, camptophyllin callus:

[0036] Preparation of medium for inducing callus: prepare one liter of MS medium with distilled water. Add 1.1 mg / L 6-benzyl adenine, 2 g / l sucrose, 3 g / l urea, 0.7 mg / l 2,4-dichlorophenoxyacetic acid, the coagulant is agar 6 g / l, and the pH is 6.0. Evenly distributed in 25 100mL Erlenmeyer flasks, stoppered, placed in a pressure cooker, sterilized at 121°C and 15 atmospheres for 20 minutes.

[0037] Select the young stems of Campylomanti radicis as explants, clean them with clear water, soak them in 70% ethanol and rinse them for 35 minutes, take them out and rinse them with clear water three t...

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Abstract

The invention belongs to the field of biotechnology, and particularly relates to a microbial transformation preparation method of hydroxycamptothecin. The invention conducts biotransformation to the camptotheca acuminata cells of a suspension culture by using non-toxic Aspergillus flavus Cr-1 strains (Aspergillus.flavus) and Rhizopus T-34 strains (Rhizopus sp.); the camptotheca acuminate cell cultures after the transformation experience separation and refining such as raw material crushing, reflex extraction, crystallization, silica gel column chromatography, crystallization, and the like, a pure hydroxycamptothecin product is obtained. The invention conducts biotransformation to the camptotheca acuminata cells of the suspension culture by using a microorganism so as to obtain a plant secondary metabolite hydroxycamptothecin which has an anti-tumor activity, wherein, the content of the plant secondary metabolite hydroxycamptothecin achieves 0.4% (g/g) of the dry weight of the cell; a new way of obtaining high-yield hydroxycamptothecin is established, thereby meeting the requirements of sustainable development; the method for separating the hydroxycamptothecin is simple, convenient and feasible; the purity of the product is high; therefore, the invention has good development prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology. In particular, it relates to a preparation method for microbial conversion of hydroxycamptothecin. Background technique [0002] Since Wall et al. in the United States first extracted Campothecin (CPT) from Campotheca involucrata, a plant unique to China in 1966, the antitumor effect of this type of alkaloid has been widely concerned. In 1985, Hsiang expounded the mechanism of anti-tumor effect of this kind of drugs, and believed that camptothecin and its derivatives take DNA topoisomerase I as the target, and inhibit the synthesis of biological DNA to exert anti-tumor effect. This makes these drugs become the second hotspot in the research of plant-based anti-tumor drugs except paclitaxel. Among them, hydroxycamptothecin (Hydroxycamptoyhecin, HCPT) and topotecan (Topotecan, TPT) and irinotecan (Camptothecin-II, CPT-II) synthesized with hydroxycamptothecin as an intermediate have shown a wide range...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12R1/67C12R1/845
Inventor 姜波王贺双高华张涓涓毕静郝爽
Owner DALIAN UNIV OF TECH
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