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Emericella nidulans for producing echinocandin B and application of emericella nidulans

A technology of echinocandins and seeds, applied in the field of echinocandin B-producing naked shells, can solve problems such as lack of industrialization value, achieve good application value, stable genetic traits, and good reproducibility Effect

Active Publication Date: 2017-03-15
ZHEJIANG HISUN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The existing echinocandin B-producing strain NRRL8112 is quite primitive, and its fermentation titer is below 1000 mg / L, so it does not have industrial value. It is urgent to transform this strain to reduce the fermentation production of echinocandin B cost, in order to be commercially available

Method used

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  • Emericella nidulans for producing echinocandin B and application of emericella nidulans
  • Emericella nidulans for producing echinocandin B and application of emericella nidulans
  • Emericella nidulans for producing echinocandin B and application of emericella nidulans

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Embodiment 1, the fermentation of bacterial strain NRRL8112 on the 250ml shaking flask

[0081] 1. Preparation of seed medium

[0082] Seed medium: glucose 10.0g / L, soluble starch 20.0g / L, hot-pressed soybean cake powder 20.0g / L, cottonseed cake powder 15.0g / L, yeast extract powder 3.0g / L, dipotassium hydrogen phosphate 3.5g / L and calcium carbonate 3.0g / L; pH 6.8 before digestion. The filling volume is 25ml / 250ml, and it is sterilized at 121°C for 30 minutes.

[0083] 2. The strain NRRL8112 was inoculated into the seed medium of the shake flask for cultivation, the cultivation temperature was 25° C., the cultivation humidity was 40-60%, the shaker amplitude was 5 cm, the shaker speed was 250 rpm, and the culture period was 63 hours.

[0084] 3. Preparation of fermentation medium

[0085] Fermentation medium: glucose 15.0g / L, maltodextrin 10.0g / L, soybean oil 50.0g / L, cottonseed cake powder 20.0g / L, soybean cake powder 10.0g / L, yeast extract powder 3.0g / L, sulfuric a...

Embodiment 2

[0095] Embodiment 2, preparation of bacterial strain 709-N5253

[0096] 1. Transfer the NRRL8112 strain to the ISP2 slant for cultivation at 28°C for 12 days, wash the slant bacteria with sterile water, put them into a small test tube with 8 glass beads for oscillation, and then filter with lens paper , to obtain the NRRL8112 bacterial suspension, and the microscopic examination of the cell dispersion reached more than 95%, so as to ensure that most of them were single cells, which could be used for subsequent mutagenesis treatment.

[0097] 2. Accurately weigh 10.0 mg of NTG (N-methyl-N'-nitro-N-nitrosoguanidine) dissolved in 1 mL of acetone, and add NRRL8112 bacterial suspension to make the final concentration reach 1.0 mg / L. Slowly shake at 4°C for 30 minutes, centrifuge to discard the supernatant, break up the precipitate with sterile saline, and repeat washing twice to terminate the reaction.

[0098] 3. Draw the suspension containing the mutagenized bacteria into a petr...

Embodiment 3

[0104] Embodiment 3, Morphological and cultural characteristics of bacterial strain 709-N5253

[0105] Refer to the relevant content in "General Mycology", "Common Bacterial System Identification Manual", "Molecular Cloning Experiment Guide" and "Chinese Pharmacopoeia" (2010 Edition) and other books to carry out the following experiments; the color judgment refers to the RAL K7 color card series Colors are compared.

[0106] Culture characteristics: using ISP1, ISP2, ISP3, ISP4, ISP5, YMS, Gaoshi No. 1, calcium malate, nutrient agar and Chase ten kinds of medium, cultured 709-N5253 obtained in Example 2 at 28°C for 5 to 7 days, Observe the color and pigment situation of mycelia, the results are shown in Table 2.

[0107] Table 2 Culture characteristics of bacterial strain 709-N5253 on 10 kinds of media

[0108]

[0109]

[0110] *Note: 0, no growth; 1, very weak growth; 2, able to grow, with a few spores; 3, good growth, with a large number of spores; 4, best growth, w...

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Abstract

The invention discloses emericella nidulans for producing echinocandin B and application of the emericella nidulans. The preservation number of the emericella nidulans is CGMCC No.10988. When the emericella nidulans for producing echinocandin B is adopted for tank fermentation, the yield of echinocandin B can be up to 4016mg / L. The emericella nidulans for producing echinocandin B, which is disclosed by the invention, is high in emericella nidulans fermentation valence, and has a very good application value.

Description

technical field [0001] The invention belongs to the field of microbial breeding and fermentation engineering, and relates to an Emericella nidulans producing echinocandin B and its application. Background technique [0002] At present, the clinically used antifungal drugs mainly include polyenes, lipopeptids, nucleosides, and azoles. Due to safety reasons, many antifungal drugs need to monitor liver and kidney functions during the course of treatment. Therefore, it is necessary to further develop a series of high-efficiency and low-toxic antifungal drugs clinically. The lipopeptide echinocandin drug that has been on the market in recent years is the most effective one. [0003] According to reports, lipopeptide antifungal drugs mainly include: Caspofungin, Anidulafungin, Micafungin, Aminocandin and the like. These antifungal drugs are all obtained by microbial fermentation to produce active intermediates, and then undergo semi-synthetic modification. Lipopeptide antifunga...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P21/04C12R1/645
Inventor 郑玲辉马海霞李娜潘玲玲徐彬
Owner ZHEJIANG HISUN PHARMA CO LTD
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