Method for converting echinocandin B into echinocandin B parent nucleus through microbial fermentation

A technology of microbial fermentation and echinocandin, applied in the field of biopharmaceuticals, can solve problems such as complicated operation, and achieve the effects of simple operation, good commercial value and high molar conversion rate

Active Publication Date: 2013-10-30
ZHEJIANG ZHENYUAN PHARMA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this me...

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1 produces the fermentation of echinocandin B deacylase bacterial strain

[0037] Species: Actinoplanes utahensis NRRL 12052

[0038] Medium formula:

[0039] Incline medium: yeast extract 0.3%, malt extract 0.3%, tryptone 0.5%, glucose 1.0%, agar 2.5%, pH 7.0-7.2, culture at 30°C for 5-7 days;

[0040] Seed medium: sucrose 2.5%, oat flour 2%, yeast powder 0.25%, K 2 HPO 4 0.1%, KCl 0.05%, MgSO 4 ·7H 2 O 0.05%, FeSO 4 ·7H 2 O 0.0002%, pH=6.8, cultivated at 30°C for 3-4 days;

[0041] Fermentation medium: 2.5% sucrose, 1.2% peanut meal powder, K 2 HPO 4 0.1%, MgSO 4 ·7H 2 O0.3%, pH=6.8, fermented at 30°C.

Embodiment 2

[0044] 1) Dissolve ECB in pure methanol at a concentration of 100 mg / ml, and after 48 hours of fermentation by the transformed bacteria, add 1.5 mg / ml of ECB substrate to start the transformation;

[0045] 2) During the conversion process, HPLC quantitatively monitors the content of the ECB mother nucleus in the fermentation broth to determine the time for adding the next batch of substrates. After 60-70% of the added ECB substrates are converted, add the next batch of substrates.

[0046] 3) After 12 hours of conversion, add ECB substrate 1.5mg / ml for the second time, and continue the conversion;

[0047] 4) After continuing to transform for 16 hours, add ECB substrate 3.0mg / ml for the third time

[0048] 5) After continuing the transformation for 24 hours, add 2.0 mg / ml of ECB substrate for the fourth time for transformation;

[0049] 6) After continuing the transformation for 24 hours, add 2.0 mg / ml of ECB substrate for the fifth time, and monitor the transformation situat...

Embodiment 3

[0052] 1) Dissolve ECB in pure ethanol at a concentration of 120 mg / ml, and after 72 hours of fermentation by the transformed bacteria, add 3.0 mg / ml of ECB substrate to start the transformation;

[0053] 2) During the conversion process, HPLC quantitatively monitors the content of the ECB mother nucleus in the fermentation broth to determine the time for adding the next batch of substrates. After 60-70% of the added ECB substrates are converted, add the next batch of substrates.

[0054] 3) After 17 hours of conversion, add ECB substrate 4.0 mg / ml for the second time, and continue the conversion;

[0055] 4) After continuing the transformation for 24 hours, add 2.0 mg / ml of ECB substrate for the third time, and monitor the transformation situation by HPLC. When the ECB substrate is exhausted, the transformation stops;

[0056] 7) Suction filtration of the fermentation broth, ECBN exists in the filtrate, and the molar conversion rate calculated by HPLC detection is 90%.

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Abstract

The invention discloses a method for converting echinocandin B into an echinocandin B parent nucleus through microbial fermentation. The method is characterized by comprising the following step of: adding an echinocandin B substrate in a fermentation process to carry out conversion, wherein the echinocandin B substrate is added in batches or added in a flowing manner.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular, the invention relates to a method for converting echinocandin B into echinocandin B mother nucleus through microbial fermentation. Background technique [0002] Both the frequency and the type of fungal infections are increasing, especially in immunosuppressed patients including HIV-infected patients, organ transplant recipients, cancer patients and those with other diseases who are undergoing immunotherapy. The main drugs currently used to treat clinical deep fungal infections are azole antifungal drugs and amphotericin B. Although these two types of drugs have played an important role in controlling clinical deep fungal infections, due to poor selectivity and drug resistance of these drugs The emergence of fungi and the lack of susceptibility to fungi such as Aspergillus and Candida albicans make the mortality rate of deep fungal infections high. Therefore, it is particularly i...

Claims

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Application Information

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IPC IPC(8): C12P21/04C12R1/045
Inventor 李继安陈详姚道明林慧敏朱秀兰张菲菲王白龙韩建栋董华成阮霞琴
Owner ZHEJIANG ZHENYUAN PHARMA CO LTD
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