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Method for separating and purifying antifungal lipopeptide

A separation and purification, anti-fungal technology, applied in the field of lipopeptide separation and purification to prevent and control plant fungal diseases, can solve the problems of complex control of process conditions, cumbersome purification steps, poor purification effect, etc., achieve good purification effect, difficult activity, separation and purification The effect of simple process

Inactive Publication Date: 2010-10-06
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

Although these methods can purify antimicrobial peptides, there are often problems such as cumbersome purification steps, complex control of process conditions, poor purification effect, and loss of activity after treatment.

Method used

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  • Method for separating and purifying antifungal lipopeptide
  • Method for separating and purifying antifungal lipopeptide

Examples

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Embodiment 1

[0026] Adopt 3 Erlenmeyer flasks of 500ml capacity, each bottle packs 100ml fermentation medium (sucrose 2%, KH 2 PO 4 0.3%, Na 2 HPO 4 1%, NH 4 NO 3 0.2%, MgSO 4 ·7H 2 O 0.02%, yeast powder 0.02%, CaCl 2 0.7μg / 100ml, MnSO 4 ·H 2 O 1μg / 100ml, pH 7.0-7.2, the balance is water), sterilized at 121°C for 30 minutes, after cooling, inoculate a ring of fresh activated antifungal lipopeptide producing bacteria marine Bacillus subtilis 3512A, and place at 28°C In the shaker, shake at 180rpm for 24 hours. After the cultivation, take it out and take it out for microscopic examination. It can be used as the seed solution.

[0027]Inoculate the cultivated seed liquid with an inoculation amount of 5% by volume in a 10L fermentation bioreactor with 6L sterilized fermentation medium inside. 5-7L / min, the stirring speed is 180-210rpm, and the aeration and stirring fermentation can be put into the tank after 46-50 hours of fermentation. This fermentation broth can be used to extract ...

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Abstract

The invention relates to separation and purification of antifungal lipopeptide of marine microorganisms, in particular to a method for separating and purifying the lipopeptide of anti-plant-fungal diseases. The method comprises the following steps that: (1) a marine bacillus subtilis3512A is adopted to conduct fermentation and culture under the suitable condition; (2) centrifugation, acid precipitation and methyl alcohol extraction are conducted after fermentation and culture, and then lipopeptide crude extract is obtained; (3) silica gel high-speed chromatography is adopted to collect active components with antifungal property; and (4) RP-HPLC is utilized to prepare and obtain an antifungal lipopeptide pure product. The invention effectively provides a method which is separated and purified from the metabolite of the marine bacillus subtilis and has anti-plant-pathogenic-fungi lipopeptide, and has the advantages of fast, simple and convenient operation, good purification effect, difficult loss of activity, low cost and the like.

Description

technical field [0001] The invention relates to the separation and purification of antifungal lipopeptides produced by marine microorganisms, in particular to a method for separating and purifying lipopeptides from marine Bacillus subtilis fermentation broth to prevent and control plant fungal diseases. Background technique [0002] Plant fungal diseases often lead to huge losses in agricultural production, ranging from reduced yields of crops to severe economic losses. However, at present, chemically synthesized pesticides are mainly used to prevent and treat fungal diseases at home and abroad. Although chemical pesticides have played a certain role in disease control, they have caused serious damage to the ecological environment during the use of pesticides, such as environmental pollution. Pesticide residues and resistance of pathogenic bacteria and so on. Therefore, looking for some environmentally friendly fungal disease control methods has been paid more and more atte...

Claims

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Application Information

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IPC IPC(8): C12P21/02C07K1/36A01P3/00C12R1/125
Inventor 徐杨王楠胡江春潘华奇刘丽王书锦
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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