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Sorangium cellulosum, preparation method and application for Phoxalone fermented by the same

A technology of Cystobacter fibroblastii and foveomycin, which can be used in fermentation, bacteria and other directions, and can solve the problems of low toxicity and unseen liver cells.

Inactive Publication Date: 2008-02-20
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Phoxalone (1,7,12,13-tetrahydroxyl-14-methoxyl-8,10-dimethyl-6-phenyl-5,15-dioxo-bicyclic [9.3.1] Pentadecane-4-one), not only is completely new in structure, but also the compound is isolated from the fermentation product of microorganisms for the first time; in terms of biological activity, it has great potential for multiple tumor cell models High anticancer activity and low toxicity to normal human liver cells, which has not been reported in the literature

Method used

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  • Sorangium cellulosum, preparation method and application for Phoxalone fermented by the same
  • Sorangium cellulosum, preparation method and application for Phoxalone fermented by the same
  • Sorangium cellulosum, preparation method and application for Phoxalone fermented by the same

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Explain how to use myxobacteria to ferment Phoxalone.

[0045] (1) Preparation of fermented seeds Inoculate an inoculum ring of S. cellulosus MWXAB-125 (So ceMWXAB-125) on a solid slant medium (g / L: α-cellulose 0.5-5.5, Angel active dry yeast 1.0 ~5.0, CaCl 2 0.3~3.0, agar 15.0, water 1000mL), cultured at 30℃ for 3~5d, inoculated in 25mL liquid seed medium (g / L: potato starch 0.5~20, glucose 0.5~10, yeast powder 0.5~10, skimmed milk powder 1.0 ~20, CaCl 2 0.3~5.0, MgSO 4 ·7H 2 O 0.3-5.0, Fe(III)-Na-EDTA 0.001-0.2, pH 7.0-7.6), cultured at 30°C, 160r / min for 3 days.

[0046] (2) The liquid composition (g / L) used in liquid fermentation is: potato starch 10.0, glucose 2.0, yeast powder 2.0, skimmed milk powder 4.0, CaCl 2 1.0, MgSO 4 ·7H 2 O 1.0, Fe(III)-Na-EDTA 0.008, XAD-16 macroporous resin 20.0, bean cake powder 10, glycerol 5.0. The medium was adjusted to pH 7.5 with 10% KOH, and sterilized at 121°C for 20 minutes.

[0047] (3) Extraction and analysis of m...

Embodiment 2

[0050] Describe how to achieve separation and purification of Phoxalone from Myxobacterium fermentation broth.

[0051] (1) Take out the XAD-16 resin in the fermentation broth by high-speed centrifugation, wash the resin with 5%-10% methanol for 2-3 times, then wash the resin with deionized water until the eluent is clear, and drain.

[0052] (2) rinse the washed resin with methanol, then concentrate the eluent to obtain a crude extract;

[0053] (3) Separation by low-pressure chromatography column: use 10.0% to 100% methanol balanced gradient elution for four times on C-18 column (10 μm × 25mm × 300mm), and use 3 column volumes of eluent for each elution, Components I, II, III, and IV were obtained respectively, and each elution time was 1 to 2 hours;

[0054] (4) Medium-pressure liquid phase separation: combine components II, III, and IV, and use 10% to 100% methanol solution for gradient elution on an AKTA rpc reverse-phase column of 15 μm×25mm×1000 mm: 10% to 30% methanol...

Embodiment 3

[0057] Describe how to realize the structure identification of focimycin (Phoxalone).

[0058] (1) HRMS (MALDI-TOF MS / MS) analysis

[0059] Spotting method: first spot 1.0 μL of the sample on the MALDI stainless steel target plate, after natural drying, then spot 0.5 μL of 0.5 g / L α-cyano-4-hydroxycinnamic acid (CCA) solution (solvent, 0.1% Trifluoroacetic acid (TFA) + 50% acetonitrile (ACN)), and dried naturally at room temperature. Another 1.0μL 0.5g / L CCA solution (no sample) was used as a blank control.

[0060] Mass Spectrometry:

[0061] The laser source is a Nd:YAG laser with a wavelength of 355nm, and the accelerating voltage is 20kV. The positive ion mode and the mode of automatic data acquisition are used to collect data. The PMF mass scan range of matrix and sample is from 100 to 1000 Da. After MS analysis, directly select ions that are different from the PMF map of the control matrix for MS / MS analysis.

[0062] MALDI-TOF-MS using reflection positive

[0063]...

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Abstract

A sorangia cellulosum and a method to prepare Phoxalone from the fermentation of the Sorangia cellulosum and the applications thereof pertain to the technical field of bio-engineering and natural medicine. The invention relates to a strain of high active anti-cancer substance - Sorangium cellulosum MWXAB-125 (So ce MWXAB-125), so as to produce Phoxalone and derivatives and analogs of Phoxalone from the fermentation of Sorangium cellulosum; the invention also relates to the preparation methods thereof, the identifications of substance structures thereof and the extrinsic anti-tumor activities as cell inhibitors.

Description

technical field [0001] The invention discloses a cellulosol and a preparation method and application thereof to ferment foliomycin, belonging to the technical fields of bioengineering and natural medicine. The present invention relates to a strain capable of producing highly active anticancer substance Sorangium cellulosum MWXAB-125 (So ce MWXAB-125), which is used to ferment and produce Phoxalone and its derivatives and Analogue, preparation method of Phoxalone, and anti-tumor application in vitro. Background technique [0002] Myxobacteria (myxobacteria) are the highest prokaryotic organisms with complex multicellular behavior and morphogenesis. They play an important role in the research of cell differentiation, development and biological evolution. Important drug-source microbial groups. According to cell morphology, myxobacteria are divided into 12 genera and about 40 species. So far, except Haploangium (Haploangium) which cannot be cultured artificially, the remainin...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/18
Inventor 陶文沂郭文杰董国秀
Owner JIANGNAN UNIV
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