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Micromonospora echinospora var. Sheshanensis C-3509 generating antitumor antibiotic calicheamicin

A calimycin, anti-tumor technology, applied in the direction of microorganism-based methods, bacteria, microorganisms, etc., to achieve the effect of high-efficiency anti-tumor antibiotics, convenient storage, and simple culture conditions

Inactive Publication Date: 2009-12-02
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, this variant has not been reported at home and abroad

Method used

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  • Micromonospora echinospora var. Sheshanensis C-3509 generating antitumor antibiotic calicheamicin
  • Micromonospora echinospora var. Sheshanensis C-3509 generating antitumor antibiotic calicheamicin
  • Micromonospora echinospora var. Sheshanensis C-3509 generating antitumor antibiotic calicheamicin

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Experimental program
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Effect test

Embodiment 1

[0038] Various Micromonospora strains isolated from the soil were respectively connected to TYM agar slant (Tryptone 5g, yeast extract 3g, maltose 10g, K 2 HPO 4 1g, KH 2 PO 41g, 18g of agar, adjusted to PH 7.0-7.2, deionized water to 1000ml), cultured at 28°C for 12 days, and then inoculated in fermentation medium (glucose 10g, sucrose 15g, yeast extract powder 3g, hairtail peptone 4g, KI 0.1g, trace element solution 1ml, cold boiled water 1000ml) 220rpm, cultured at 28°C for 3 days, centrifuged at low temperature to get the supernatant, detected by spermatogonial cell method, and screened the positive bacterial strain; take the fermentation liquid of the bacterial strain, The ultraviolet absorption spectrum was measured, and the strain C-3509 similar to calimycin was screened out.

Embodiment 2

[0039] Classification of Micromonospora C-3509

[0040] Classification culture adopts the usual representative culture medium and the culture medium proposed by the International Streptomyces Planning Conference (abbreviated as ISP medium), and cultured statically at 28°C for 21 days to observe the culture characteristics of the C-3509 strain. The color identification refers to the Ridgwa map, the physiological and biochemical experiments refer to the literature methods, the chemical composition of the cell wall refers to the methods of Backer, Lechevalier and Hasegawa, and the analysis of phospholipids, quinones and G+C mol% follows Ross and Ruan Jisheng from the Institute of Microbiology, Chinese Academy of Sciences. method to proceed.

[0041] 1. Morphological characteristics of bacteria

[0042] C-3509 strain has bare, smooth or rough colonies on various agar media, and does not form aerial hyphae.

[0043] Vegetative mycelium is generally light pink orange, apricot ye...

Embodiment 3

[0060] Fermentation of Micromonospora strain C-3509

[0061] The slant of C-3509 uses TYM medium (Tryptone 5g, yeast extract 3g, maltose 10g, K 2 HPO 4 1g, KH 2 PO 4 1g, 18g of agar, adjust the pH to 7.0-7.2, and make 1000ml of deionized water). Inoculate C-3509 on the slant medium and culture at 28°C for 12 days, so that the slant is covered with abundant black spores.

[0062] Dig the well-grown slant culture and inoculate it into the primary seed medium (dextrin 15g, Malt Extract 10g, Proteose Peptone 5g, Yeast extract 5g, adjust the pH to 7.0, make 1000ml of deionized water), and culture with shaking at 28°C for 96 hours .

[0063] The primary seed is transferred to the secondary seed medium (dextrin 15g, glucose 10g, Yeast extract 3g, hairtail peptone 5g, pH 7.0, deionized water to 1000ml), inoculum size is 5%, 28 ℃ shaking culture for 48 hours .

[0064] Secondary seeds were inoculated in fermentation medium (glucose 10g, sucrose 15g, yeast extract powder 3g, ...

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Abstract

The invention relates to Micromonospora echinospora var. Sheshanensis C-3509 generating enediyne antitumor antibiotics. Experimental results show that the fermentation liquor of the Micromonospora echinospora var. Sheshanensis C-3509 is tested positive by a spermatogonium method, and is subjected to isolation and purification to obtain high-efficiency antitumor bioactive substances, namely four compositions of Calicheamicins gamma1, gamma1, beta1 and beta1; therefore, the Micromonospora echinospora var. Sheshanensis C-3509 is hopeful to be used for preparing the enediyne antitumor antibiotics.

Description

Technical field: [0001] The invention relates to a Micromonospora aculeatus varietal C3509 that produces enediyne anti-tumor antibiotic calimycin and is screened from Chinese soil. Background technique: [0002] Since the middle of this century, the incidence of certain malignant tumors has doubled almost every ten years, and has become a common and frequently-occurring disease that seriously threatens people's lives. The prevention and treatment of tumors has always been a hot spot in medical research. Chemotherapy has played an irreplaceable role since nitrogen mustard alkylating agents were used in tumor treatment in the 1950s. However, in order to meet urgent clinical needs, researchers have to intensify efforts, expand the depth and breadth of research, and constantly open up ways to find new chemotherapeutic drugs. [0003] Microorganisms widely exist in nature, and there are many kinds of them, and the number of their metabolites is also huge. Over the years, micro...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/56C12R1/29
Inventor 邵荣光甄永苏胡继兰张文军赵春燕杨秀萍李电东
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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