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Microbe generating antifungal lipopeptid and preparation method and application of antifungal lipopeptid thereof

An antifungal and microbial fermentation technology, applied in the biological field, can solve problems such as drug resistance of pathogenic fungi

Inactive Publication Date: 2010-11-24
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current medicine for the treatment of skin ringworm is the ointment containing ketoconazole or clotrimazole, which belongs to chemically synthesized drugs, and long-term use will lead to drug resistance of pathogenic fungi

Method used

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  • Microbe generating antifungal lipopeptid and preparation method and application of antifungal lipopeptid thereof
  • Microbe generating antifungal lipopeptid and preparation method and application of antifungal lipopeptid thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0061] (1) Cultivation of microbial strains and fermentation of antifungal lipopeptides

[0062] [Broth medium (g / L): peptone 10.0, beef extract 5.0, NaCl 5.0, pH 7.2; solid medium needs to add 20.0 agar.

[0063] YEPD medium (g / L): peptone 20.0, yeast powder 10.0, glucose 20.0; solid medium needs to add 20.0 agar. ] Here, the known content can be deleted.

[0064] The composition of the slant and seed medium: (g / L): glucose 3, tryptone 5, yeast extract powder 6, NaCl6, deionized water preparation, pH 7.5; solid medium needs to add 15g agar powder.

[0065] Composition of fermentation medium (g / L): 5 g of sugary material (glucose), 15 g of nitrogen source (ammonium nitrate), 1 g of sodium citrate, MgSO 4 .7H 2 0, 0.5g, K 2 HPO 4 2~6g, KH 2 PO 4 5g, pH 7.0-7.2. The sterilization temperature of the seed medium and the fermentation medium is 115°C for 20 minutes.

[0066] The strain BI1 was inoculated from a slant into a 250 mL Erlenmeyer flask filled with 50 mL of seed ...

Embodiment 2

[0068] (1) The fermentation strain used is CGMCC No.3412, and the composition of the fermentation medium is (g / L): glucose 10g, nitrogen source 20g, sodium citrate 3g, MgSO 4 ·7H 2 0, 0.2g, K 2 HPO 4 2g, KH 2 PO 4 3g, pH 7.0;

[0069] (2) Fermentation culture: Insert the cultured seeds into the fermentation medium at an inoculum size of 2% (v / v), the culture temperature is 30°C, and the fermentation ventilation volume is 5m 3 / h, tank pressure 0.03MPa, stirring speed 80 rev / min, fermentation time 30 hours, feed sterile air during fermentation, maintain the pH of the fermentation broth at 7.5 with ammonia water, the sterilization temperature of the seed medium and the fermentation medium is 121°C, 20min. The output of antifungal lipopeptide in the fermentation broth was 1.5g / L.

Embodiment 3

[0071] (1) The fermentation strain used is CGMCC No.3412, and the composition of the fermentation medium is (g / L): corn flour 35, nitrogen source peptone 15, sodium citrate 1, MgSO 4 ·7H 2 0, 0.8, K 2 HPO4 4 , KH 2 PO 4 6, pH 7.2.

[0072] (2) Fermentation culture: Insert the cultured seeds into the fermentation medium at an inoculum size of 2% (v / v), the culture temperature is 30°C, and the fermentation ventilation volume is 5m 3 / h, tank pressure 0.05Pa, stirring speed 60 rpm, fermentation time 45h, feed sterile air during the fermentation process, maintain the pH of the fermentation broth with ammonia water at 7.2, and the output of antifungal lipopeptide in the fermentation broth is 1.4g / L .

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Abstract

The invention relates to a microbe generating antifungal lipopeptid and a preparation method and application of the antifungal lipopeptid thereof, wherein a strain generating the antifungal lipopeptid through microbial fermentation is classified to belong to Bacillus subtilis BI1, and the collection number is CGMCC No. 3412. The invention also relates to a preparation method of fermented antifungal lipopeptid, comprising the following steps of: inoculating cultured seeds into a fermentation culture medium according to 1%-10% (v / v) of inoculation quantity by adopting fermentation strain BI1, wherein culture temperature is 30-40 DEG C, fermentation ventilatory capacity is 5-13 m<3> / h, tank pressure is 0.02-0.09 MPa, stirring speed is 100-150 turns / minute, and fermentation time is 30-60 hours; introducing sterile air in the fermentation process; and maintaining the pH of fermentation liquor between 6.5 and 7.5 by using ammonia water or liquid caustic soda, wherein the output of the antifungal lipopeptid in the fermentation liquor is 1-2 g / L. The invention also discloses the application of the antifungal lipopeptid in preparing drugs treating dermatophytosis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to microorganisms producing antifungal lipopeptides and preparation methods and applications of antifungal lipopeptides. More specifically: a microbial fermentation to produce antifungal lipopeptide strains, which is a new strain found in the present invention that can effectively produce antifungal lipopeptides, and can be used to ferment and produce antifungal lipopeptides. Effective for the treatment of skin ringworm. Background technique [0002] Lipopeptide is a secondary metabolite produced by Gram-positive bacillus. The molecule of lipopeptide is composed of two parts: hydrophilic peptide bond and lipophilic aliphatic hydrocarbon chain. Due to its special chemical composition and amphiphilic molecular structure, it has the characteristics of biosurfactant. It is widely used in medicine, agriculture, food and cosmetics. It has important application prospects in fields such as oil ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00A61K38/16A61P17/00A61P31/10C12R1/125
Inventor 王德培徐进
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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