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Sorangiumcellulosum strain and application thereof to synthesis of epothilone

The technology of a kind of epothilone and bacterial strain is applied in the application field of epothilone synthesizing aspect of the bacterial strain, and achieves the effect of huge economic benefit and market value

Active Publication Date: 2015-04-01
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the chemical modification method must also rely on the supply of a large amount of raw materials before it can be implemented.

Method used

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  • Sorangiumcellulosum strain and application thereof to synthesis of epothilone
  • Sorangiumcellulosum strain and application thereof to synthesis of epothilone
  • Sorangiumcellulosum strain and application thereof to synthesis of epothilone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Isolation method of strain So2163

[0025] Grind the soil sample and evenly sprinkle it on the CNST medium, observe it after cultivating at 30°C for 7-10 days, and pick and inoculate it into a new CNST medium for purification and storage after the initial growth of bacteria.

[0026] CNST medium formula ( / L): KNO 3 0.5g / L; Na 2 HPO 4 0.25g / L; MgSO 4 1g / L; pH 7.5; add 1.2% agar powder to the solid medium, and spread sterilized filter paper sheets as carbon source; sterilize with moist heat at 102.34 kPa for 30min and set aside.

Embodiment 2

[0028] Species Identification of Strain So2163

[0029] (1) Morphological method identification

[0030] The strain So2163 was cultured by CNST at 30°C for 5 days, and microscopic examination was carried out after the colony expanded sufficiently and the fruiting body structure was formed, and the bacterial cells, fruiting body shape and expanded bacterial film shape were observed respectively (see the appendix for the experimental results. figure 1 ), the results showed that the strain possessed typical S. cellulosus ( Sorangium cellulosum ) Morphological features.

[0031] (2) Identification by molecular biology methods

[0032] By designing 16S rDNA bacterial universal amplification primers 27F (5'-AGA GTT TGA TCC TGG CTC AG-3') and 1492R (5'-TAC CTT GTT ACG ACT T-3'), PCR was carried out using the genome of strain So2163 as a template Amplified and obtained a 16S rDNA fragment with a length of 1466bp, compared with the sequence in GeneBank after sequencing, it was foun...

Embodiment 3

[0035] Cultivation method of bacterial strain So2163 and pretreatment process of fermentation product

[0036] EPM medium is used to synthesize epothilone, the culture temperature is 30°C, the rotation speed is 200rpm, and the culture time is 7-8 days. After the fermentation, the XAD-16 macroporous adsorption resin is collected, washed 2-3 times with distilled water, and then used for 2-fold volume chromatography Extract with pure methanol for 2 hours, centrifuge at 10,000 rpm for 10 minutes, and obtain the fermented crude extract.

[0037] EPM medium formula ( / L): 2.0 g potato starch, 2.0 g glucose, 2.0 g bean cake powder; 1.0 g skimmed milk, MgSO 4 1.0 g, CaCl 2 1.0 g, VB 12 0.5 mg, Amberlite XAD-16 macroporous adsorption resin 2% (v / v), adjust the pH to 7.2, and sterilize at 121°C for 20 minutes.

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Abstract

The invention belongs to the fields of bioengineering and biological pharmacy, and in particular to a Sorangiumcellulosum So2163 strain, and application of the strain to synthesis of epothilone. The Sorangiumcellulosum So2163 strain was preserved on June 10, 2012 in China Center for Type Culture Collection, and has a collection number CCTCC M 2012211. The Sorangiumcellulosum So2163 strain involved in the invention provides new strain resource for industrialization production of epothilone.

Description

technical field [0001] The invention belongs to the fields of bioengineering and biopharmaceuticals, and in particular relates to fibrocystic bacterial strains Sorangium cellulosum So2163 also relates to the application of this strain in the synthesis of epothilones. Background technique [0002] Cancer is a common and frequently-occurring disease that seriously threatens human health and life. It is an important scientific research topic in the field of medical science in all countries in the world. At present, there is no satisfactory treatment. Paclitaxel is currently the most widely used antineoplastic drug, with an annual sales volume of US$2 billion, accounting for 10% of the antineoplastic drug market. Due to its anti-cancer general expansion and good treatment prospects for non-tumor diseases, it is estimated that the world demand will reach 2,000 kilograms per year in the future, and the price of paclitaxel preparations from the US company BMS is 5-10 million ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/18C12R1/01
Inventor 赵林刘新利李越中李鹏飞孙欣
Owner QILU UNIV OF TECH
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