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Separating and extracting method of Bacillus subtilis lipopeptide antibiotic substance

A Bacillus subtilis, extraction method technology, applied in the field of separation and extraction of Bacillus subtilis lipopeptide antibacterial substances, can solve the problems of high equipment requirements, unsuitable for large-scale production, complicated operation, etc. Spectrum antifungal activity, simple and rapid preparation process

Inactive Publication Date: 2009-02-25
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

The prior art extraction method is complex in operation and requires high equipment requirements, and can be used for a small amount of preparation of lipopeptide antibacterial substances, but is not suitable for large-scale production

Method used

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  • Separating and extracting method of Bacillus subtilis lipopeptide antibiotic substance

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Experimental program
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Embodiment Construction

[0015] Preparation of crude extract of Bacillus subtilis fermentation broth:

[0016] Bacillus subtilis was inoculated in LB medium, and cultured on a shaker at 37° C. at 200 rpm for 24 hours to obtain a Bacillus subtilis fermentation broth. Described LB culture medium: peptone 1g, yeast powder 0.5g, NaCl1g, water 100mL, pH7.2-7.4.

[0017] Take 50 mL of the Bacillus subtilis fermentation broth prepared above, centrifuge at 6,000 rpm for 10 minutes, take the supernatant, add 100 mL of acetone to the supernatant, precipitate overnight at 4°C, and collect the supernatant by centrifugation at 12,000 rpm , concentrated to 5 mL by rotary evaporation at 60°C, and freeze-dried to obtain 200 mg dry matter of the crude extract of the fermentation broth, which was set aside.

[0018] Weigh 40g of Sephdex G-25 gel, fully swell, and routinely pack the column with a bed volume of 100mL (φ1.7cm×44cm). The crude extract of the lyophilized fermentation broth obtained above is formulated into...

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Abstract

The invention relates to the field of biotechnology, in particular to a method for separating and extracting bacillus subtilis lipopeptid antibacterial material. The method more particularly comprises the steps Sephadex G-25 column chromatography is carried out on crude extract of fermentation liquor of the bacillus subtilis, active peak is collected by chromatography, and the Sephadex G-25 column chromatography is carried out on the active peak which is collected by Sephadex G-25 column; the active peak is collected, and the obtained substance is seven lipopeptid antibacterial materials after being detected by mass spectrum. Furthermore, the extracted lipopeptid antibacterial material has good inhibition effect on multiple pathogenic fungis, and has the advantages of stable temperature and pH value, prolease hydrolysis resistance, etc.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for separating and extracting bacillus subtilis lipopeptide antibacterial substances. technical background [0002] Bacillus subtilis can produce a variety of antibacterial substances, and lipopeptide antibacterial substances are one of the most important types. They have strong antibacterial activity. These antibacterial substances are stable in nature and have good resistance to high temperature, acid-base, and protease. Receptivity, and low toxicity, low residue, has potential application value in biological pesticides, feed additives, food additives, antifungal drugs, antiviral drugs, etc., and has become a hot research topic at home and abroad. [0003] Bacillus subtilis lipopeptide antibacterial substances are usually divided into three groups due to their different structures: iturins, surfactin and plipastatin-fengycin family antibacterial substances, iturins are pep...

Claims

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Application Information

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IPC IPC(8): C07K1/16C12P21/00C12R1/125
Inventor 李旭张惠文苏振成张成刚
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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