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A method for fermenting and producing bafilomycin a1

A fermentation strain and fermentation culture technology, applied in the field of fermentative production of bafilomycin A1 and fermentative production of bafilomycin A1 by Streptomyces griseus, can solve the problems of low fermentation titer and meet the needs of industrialization , improve the yield, and benefit the effect of extraction and purification

Active Publication Date: 2021-02-02
FUJIAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] For this reason, the technical problem to be solved by the present invention is to provide a method for fermentative production of bafilomycin A1, to solve the problem of low fermentation titer of bafilomycin A1 compounds in the prior art

Method used

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  • A method for fermenting and producing bafilomycin a1
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  • A method for fermenting and producing bafilomycin a1

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1 strain source

[0031] In August 2012, our laboratory obtained several strains of actinomycetes in the sediments of the East China Sea through isolation, cultivation, fermentation, and activity determination. After preliminary determination, one strain of Streptomyces had strong antibacterial activity against Aspergillus niger .

[0032] The compound Ba0115 was obtained through natural separation and purification, fermentation and post-extraction process of the strain, which was identified as homogeneous to bafilomycin A1 by mass spectrometry and NMR. However, the fermentative production of bafilomycin A1 by the original strain has a low titer. In this embodiment, this strain is used as the starting strain for mutagenesis.

[0033] Preparation of spore suspension: inoculate the starting strain on the ISP2 slope at 28°C for about 10 days, take the fresh slope and add 10 mL of sterile saline, gently scrape it off with an inoculation spatula, pour it into a ster...

Embodiment 2

[0039] Morphological and culture characteristics of embodiment 2 Streptomyces FIM-Ba150115

[0040] After identification, Streptomyces FIM-Ba150115 described in this example was positive by Gram staining. After growing for 7 days on the culture mediums such as Gaoshi Synthetic No. 1 Agar and ISP2, the hyphae in the base were well developed, without septa, and without Fractured, aerial hyphae grow well, multi-branched, spore filaments are straight or flexible.

[0041] Seven kinds of media including ISP1, ISP2, ISP3, and ISP4 were used respectively, and after culturing at 30°C for 14 days, the color and pigment of mycelium were observed. The results are shown in Table 2 below.

[0042] Table 2 Culture characteristics of FIM-Ba150115

[0043] culture medium growing situation aerial hyphae base hyphae Soluble pigment ISP1 ++ gray grey none ISP2 +++ taupe taupe none ISP3 + Ash poor none ISP4 + light gray poor none ...

Embodiment 3

[0048] The detection of bafilomycin A1 in the fermented liquid of embodiment 3

[0049] In the following examples of the present invention, the detection of the content of bafilomycin A1 in the fermentation broth is carried out according to the HPLC method in the prior art.

[0050] Fermentation sample preparation: Take 10 mL of the fermentation liquid of the strain, add 20 mL of methanol and mix, oscillate and mix for 20 min, centrifuge, set aside, and use HPLC method to detect bafilomycin A1.

[0051] High performance liquid chromatography analysis conditions Chromatographic column detection conditions include:

[0052] Ultimate LP-C18 analytical column (5μm, 4.6×250mm);

[0053] Mobile phase: methanol-water solution with a volume ratio of 85:15;

[0054] Flow rate: 1.0mL / min;

[0055] Injection volume: 10μL;

[0056] Detection wavelength: 247nm;

[0057] Use time: 30min;

[0058] The content was calculated using the peak area.

[0059] Scan the bafilomycin A1 referen...

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Abstract

The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for fermenting and producing bafilomycin A1, and more particularly relates to a method for fermenting and producing bafilomycin A1 with Streptomyces grisea. The present invention obtains a high-yield bafliomycins compound Streptomyces griseus through strain screening and mutation breeding screening, the strain can effectively improve the potency of the bafliomycin A1 compound in the fermentation broth, and through the fermentation process of the fermentation medium and fermentation Careful study of the conditions has effectively improved the efficiency of fermentation and production of bafliomycin A1, and increased the yield of the target compound bafliomycin A1. Fermentation experiments show that the titer of bafliomycin A1 produced by the self-selected Streptomyces griseus FIM‑Ba150115 fermented by the present invention is as high as 615.87mg / L, which greatly facilitates the extraction and purification of bafliomycin A1 and can meet the needs of industrialization.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for fermenting and producing bafilomycin A1, and more particularly relates to a method for fermenting and producing bafilomycin A1 with Streptomyces grisea. Background technique [0002] Bafliomycins are a class of polyene macrolide antibiotics with a 16-membered ring skeleton produced by the fermentation of actinomycetes. The chemical structure is very special. Most of these compounds contain a six-membered ring hemicondensation Aldehyde structure. The reported bafilomycins compounds include bafilomycin A1, A2, B1, B2, C1, C2, D, E, F, K and L, etc. Bafilomycins have various biological activities such as antibacterial, antifungal, insecticidal, antitumor and immunosuppressive, and can also be used to treat osteoporosis and deafness. And existing studies have shown that Bafilomycins series compounds, especially bafilomycinA1, have specific au...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/16C12N1/20C12R1/465
CPCC12P17/162C12N1/205C12R2001/465
Inventor 周剑江红
Owner FUJIAN INST OF MICROBIOLOGY
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