Method and bacterial strain for fermentation preparation of Echinocandin B

A technology for seeds and Aspergillus, applied in the field of preparation of echinocandin B, can solve the problems of low unit yield and long fermentation period, and achieve the effects of convenient cultivation, efficient production and good application prospects

Active Publication Date: 2014-02-05
ZHEJIANG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0008] The purpose of the present invention is to provide a high-yielding strain of echinocandin B and a method for producing echinocandin B by fermen

Method used

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  • Method and bacterial strain for fermentation preparation of Echinocandin B
  • Method and bacterial strain for fermentation preparation of Echinocandin B
  • Method and bacterial strain for fermentation preparation of Echinocandin B

Examples

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Example Embodiment

[0024] Example 1: Aspergillus nidulans mutant strain CCTCC NO: M 2012300 fermentation to produce echinocandin B

[0025] The spore suspension of the mutant strain of strain Aspergillus nidulans CCTCC NO:M 2012300 was inoculated into a 250ml Erlenmeyer flask containing 50.0ml seed culture medium and cultured at 25°C for 48h to obtain seed liquid. Seed medium preparation: glucose 1.0%, glycerol 2.5%, cottonseed powder 5%, solvent is distilled water, distilled water 1000.0ml, pH value is natural, and sterilized at 121°C for 20 minutes.

[0026] In order to investigate the influence of the compound nitrogen source of the medium on the fermentation level of Echinocandin B, five different fermentation mediums were designed, the volume of shake flask was 150mL / 500mL, and the composition of the fermentation medium were:

[0027] Medium I: glycerol 1%, peptone 1%, L-proline 0.5%, mannitol 10%, soybean meal 10%, peanut oil 2%, K 2 HPO 4 0.8%, MgSO 4 7H 2 O 0.05%, MnPO 4 H 2 O0.01%, FeSO 4 7H 2...

Example Embodiment

[0035] Example 2: Preparation of echinocandin B by fermentation of Aspergillus nidulans ZJB09223 mutant strain

[0036] The spore suspension of Aspergillus nidulans ZJB09223 mutant strain Aspergillus nidulans ZJB09223 was inoculated into a 250ml Erlenmeyer flask containing 50.0ml seed culture medium, cultured at 25°C for 48h to obtain seed liquid. Seed culture medium preparation: glucose 1.0%, glycerol 2.5%, cottonseed powder 5%, solvent is distilled water, distilled water 1000.0ml, pH value is natural; seed culture medium is sterilized at 121℃ for 20 minutes.

[0037] In order to investigate the effect of the compound carbon source of the medium on the fermentation level of echinocandin B, five different fermentation mediums were designed, the volume of shake flask was 150mL / 500mL, and the composition of the fermentation medium were:

[0038] Medium I: glycerol 0.5%, peptone 3%, L-proline 0.5%, mannitol 10%, soybean meal 5%, peanut oil 2%, K 2 HPO 4 0.8%, MgSO 4 7H 2 O 0.05%, MnPO 4...

Example Embodiment

[0046] Example 3: Preparation of Echinocandin B by Aspergillus nidulans ZJB09223 mutant strain

[0047] The spore suspension of Aspergillus nidulans ZJB09223 mutant strain Aspergillus nidulans ZJB09223 was inoculated into a 250ml Erlenmeyer flask containing 50.0ml seed culture medium, cultured at 25°C for 48h to obtain seed liquid. Seed medium preparation: glucose 1.0%, glycerol 2.5%, cottonseed powder 1-10%, solvent is distilled water, distilled water 1000.0ml, pH value is natural; seed medium is sterilized at 121°C for 20 minutes.

[0048] In order to investigate the influence of medium inorganic salt ions on the fermentation level of echinocandin B, five different fermentation mediums were designed, the volume of shake flask was 150mL / 500mL, and the composition of the fermentation medium were:

[0049] Medium I: glycerol 1%, peptone 1%, L-proline 0.5%, mannitol 6%, soybean meal 5%, peanut oil 2%, K 2 HPO 4 0.8%, MgSO 4 7H 2 O 0.05%, MnPO 4 H 2 O0.01%, FeSO 4 7H 2 O 0.005%, CaCl 2 ...

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Abstract

The invention provides a strain of Aspergillus nidulans mutant, Aspergillus nidulans ZJB09223, and a fermentation method for preparing Echinocandin B by using the strain. The strain is preserved in China Center for Type Culture Collection with address of Wuhan University, Wuhan, China and zip code of 430072, and has a preservation number of CCTCC NO:M 2012300, and the collection date is 25, July, 2012. The invention employs a LiCl / UV / sodium nitrite compound mutation combined with resistance screening to obtain the Aspergillus nidulans mutant ZJB09223; the mutant strain is capable of efficient production of Echinocandin B, and is convenient for culture. The preparation process of Echinocandin B is simple and efficient, can obtain high-purity Echinocandin B and has good application prospects.

Description

(1) Technical field [0001] The invention relates to a mutant strain of Aspergillus nidulans—Aspergillus nidulans (Aspergillus nidulans) ZJB09223, and a method for preparing echinocandin B by using the fermentation method of the Aspergillus nidulans ZJB09223. (2) Background technology [0002] Echinocandin B (Echinocandin B) is a natural cyclic lipopeptide compound with a cyclic hexapeptide core and fatty acid side chain structure. The chemical name is and the molecular formula is C 52 h 81 N 7 o 16 , the molecular weight is 1060, and the structural formula is as follows: [0003] [0004] In recent years, due to the extensive use of immunosuppressants due to organ transplantation, the application of chemotherapy and more aggressive medical methods and other reasons, the number of immunocompromised patients has increased, and the incidence of fungal infections has increased significantly, especially deep fungal infections. Morbidity and mortality are increasing year b...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P21/04C12R1/66
Inventor 郑裕国邹树平牛坤钟伟毛健沈寅初
Owner ZHEJIANG UNIV OF TECH
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