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The co-culture method of sphingomonas sp. bacterial strain and aspergillus sp. fungus strain, new anti-cancer and antibiotic glionitrins derived from this co-culture method, and pharmaceutical composition containing glionitrins or pharmaceutically ac

A technology of Sphingomonas and Aspergillus, which is applied in the directions of methods of using bacteria, methods of using fungi, drug combinations, etc., and can solve the problems of spending a lot of time, money and labor.

Inactive Publication Date: 2010-10-20
KOREA INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to improve the efficiency of developing new biologically active substances from nature, the cost of a large amount of time, money and labor in repeated isolation processes from raw materials is a major obstacle

Method used

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  • The co-culture method of sphingomonas sp. bacterial strain and aspergillus sp. fungus strain, new anti-cancer and antibiotic glionitrins derived from this co-culture method, and pharmaceutical composition containing glionitrins or pharmaceutically ac
  • The co-culture method of sphingomonas sp. bacterial strain and aspergillus sp. fungus strain, new anti-cancer and antibiotic glionitrins derived from this co-culture method, and pharmaceutical composition containing glionitrins or pharmaceutically ac
  • The co-culture method of sphingomonas sp. bacterial strain and aspergillus sp. fungus strain, new anti-cancer and antibiotic glionitrins derived from this co-culture method, and pharmaceutical composition containing glionitrins or pharmaceutically ac

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0122] Embodiment 1: screening and selected bacterial strains and fungal strains for co-cultivation

[0123] The present inventors collected water of pH 3.0 from the inside of Imgok Mine (Gangneung-si, Gangwondo, Korea), and performed centrifugation to obtain a precipitate. The precipitate was suspended and diluted in saline, and the diluted precipitate was inoculated on YM agar medium (YMA) and Capex-Dox agar medium (CDA), followed by culturing at 25° C. for 10 days. A single strain was isolated, resulting in 300 strains. Among the obtained strains, selected bacterial strains and fungal strains for co-cultivation were inoculated on Capex-Dox liquid medium, and then cultured in a shaking incubator at 25° C. for 5-7 days. Chromosomal DNA of the resulting strains was isolated and 16S rDNA sequencing was performed to identify the strains. As a result, the strain KMK-001 had 98.0% homology with Sphingomonas A1XXyl1-5, which indicated that it was a novel Sphingomonas strain. K...

Embodiment 2

[0125] Embodiment 2: Other of novel Sphingomonas KMK-001 and Aspergillus strain KMC-901 to cultivate

[0126] The inventors inoculated the novel bacterial strain KMK-001 and the fungal strain KMC-901 in Capex-Dox liquid medium respectively, and then cultured them in a shaking incubator at 25°C for 3 days. The two strains were cultured in a 1 L Erlenmeyer flask containing 0.5 L of Capex-Dox broth for mass production. Two days later, 250 μL of the culture solution of the fungal strain KMC-901 was added to the culture solution of the Sphingomonas strain KMK-001, followed by co-cultivation. The co-culture containing the above strains was observed under a microscope. Such as Figure 5 As shown, the bacterial strains together with the fungal strains are shown around the mycelium (see Figure 5 ).

[0127] The production of new compounds in the co-cultures was studied by HPLC. Carry out HPLC under the following conditions (apparatus: Agilent 1100LC / MS system; Elution rate: 0...

Embodiment 3

[0128] Example 3: Isolation and purification of glionitrin

[0129] The present inventors extracted organic compounds from the culture solution obtained in Example 2 using ethyl acetate. The extract was dried at 30° C. under reduced pressure using a rotary vacuum evaporator, and then the obtained extract was subjected to reverse-phase column chromatography, resulting in 6 fractions. Elution was performed with water and acetonitrile. More precisely, elution was started with 20% acetonitrile / water and the amount of acetonitrile was increased by 20%. Use 100% methanol as the final solvent. As a result of analyzing the obtained fractions by HPLC, it was confirmed that the novel glionitrin was contained in the 60% acetonitrile / water fraction. The fractions were purified by normal phase liquid chromatography under isocratic conditions of 90% dichloromethane / ethyl acetate and 60% dichloromethane / ethyl acetate using ethyl acetate and dichloromethane as solvents . Glionitrin A w...

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Abstract

The present invention relates to a co-culture method of Sphingomonas sp. bacterial strain and Aspergillus sp. fungus strain, in which the novel Sphingomonas sp. bacterial strain KMK-001 is cultured in a liquid medium and the novel Aspergillus sp. strain KMC-901 separately cultured in another liquid medium is added to the above culture solution, a novel glionitrin biosynthesized therefrom and a pharmaceutical composition comprising the said glionitrin or its pharmaceutically acceptable salt as an active ingredient. The glionitrin herein has strong cytotoxic effect on cancer cells and has antibiotic effect on 10 pathogenic bacteria including the novel Sphingomonas sp. bacterial strain KMK-001, so that it can be effectively applied in antibiotics or anti-cancer agents.

Description

technical field [0001] The present invention relates to a method for co-cultivating bacterial strains of the genus Sphingomonas (Sphingomonas sp.) and fungal strains of the genus Aspergillus (Aspergillus ap.), wherein the novel bacterial strain KMK-001 of the genus Sphingomonas is cultivated in a liquid In the medium, and to the above-mentioned culture medium, add the novel Aspergillus strain KMC-901 that is separately cultivated in another liquid medium. The present invention also relates to a novel glionitrin biosynthesized by the above method, and a pharmaceutical composition comprising the glionitrin or a pharmaceutically acceptable salt thereof as an active ingredient. Background technique [0002] In order to obtain new substances from natural sources, several studies have been carried out. However, spending a lot of time, money and labor in repeated isolation from raw materials is a major obstacle to improving the efficiency of developing new substances with biologic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L29/00
CPCC12N1/20C12R1/01C12P39/00C12P17/185A23L1/3014A23L1/0345C12P17/182A23L1/30C12R1/66C12N1/14A23L29/065A23L33/10A23L33/135A61P31/04A61P35/00C12N1/145C12N1/205C12R2001/01C12R2001/66C12N1/00C12N1/16
Inventor 梁贤玉权学哲朴贤峰柳智慧
Owner KOREA INST OF SCI & TECH
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