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A kind of preparation method of iturin A

A technology for iturin and bacteria-producing bacteria, which is applied in the field of separation and extraction of iturin A, can solve the problems of inability to perform filtration operation, complicated operation and low efficiency, etc., so as to improve the possibility of popularization and application and improve the processing efficiency. , the effect of simple process route

Inactive Publication Date: 2011-12-21
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This is because the efficiency of the traditional filtration sterilization method is low, and the labor intensity is high. Even when filtration separation occurs, due to the small diameter of the bacteria, it is easy to form a sticky filter cake with proteins, sugars, etc. to block the filter, making the filtration operation difficult. Impossible
However, using centrifugation to remove bacteria and protein can achieve the desired effect, but the equipment is expensive, the operation is complicated, the investment is large, the energy consumption is high, and the production cost of the factory is high.

Method used

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  • A kind of preparation method of iturin A

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Effect test

Embodiment 1

[0024] Use 3 250mL Erlenmeyer flasks, each containing 100mL medium A (glucose 2.0%, peptone 3.0%, beef extract 0.5%, magnesium sulfate 0.35%, potassium dihydrogen phosphate 0.3%), sterilize at 120°C for 30 minutes, after cooling The activated Bacillus subtilis ZK-1 strain capable of producing Iturin A and its homologues was inoculated, placed at a temperature of 30° C., and shaken for 18 hours as a seed solution. Use 10 1000mL Erlenmeyer flasks, each containing 300mL medium A (2.0% glucose, 3.0% peptone, 0.5% beef extract, 0.35% magnesium sulfate, 0.3% potassium dihydrogen phosphate), and sterilize at 120°C for 30 minutes. The seed solution is inoculated with an inoculum amount of 5%-10%, placed at a temperature of 30° C., shaken for 72 hours-120 hours, and fermented at a pH of 6-8.

[0025]After the fermentation is finished, take 2.0L fermented liquid and add water with 200% of the fermented liquid volume to dilute, adjust the pH to 6.0-9.0 with 15% hydrochloric acid solution...

Embodiment 2

[0029] Use 3 250mL Erlenmeyer flasks, each containing 100mL medium A (glucose 2.0%, peptone 3.0%, beef extract 0.5%, magnesium sulfate 0.35%, potassium dihydrogen phosphate 0.3%), sterilize at 120°C for 30 minutes, after cooling The activated Bacillus subtilis ZK-1 strain capable of producing Iturin A and its homologues was inoculated, placed at 30° C., and shaken for 18 hours. Inoculate the cultured strain solution into 3L sterilized medium B (medium B: starch 2.0%, lactose 1.0%, sucrose 1.0%, glycerol 0.5%, glucose 1.0%, 1.0% of peptone, 5.0% of peanut powder, 2.0% of soybean powder hydrolyzate, 3.0% of bionitrogen, 0.1% of potassium dihydrogen phosphate, 0.5% of sodium chloride, and 0.5% of ferric sulfate). Under the temperature of 28°C-30°C, aerated and stirred for 72 hours-120 hours, the fermentation is controlled at pH 6-8.

[0030] After the fermentation is finished, take 2.0L of fermentation broth and add water with 100% of the volume of the fermentation broth to dilu...

Embodiment 3

[0034] Use 3 250mL Erlenmeyer flasks, each containing 100mL medium A (glucose 2.0%, peptone 3.0%, beef extract 0.5%, magnesium sulfate 0.35%, potassium dihydrogen phosphate 0.3%), sterilize at 120°C for 30 minutes, after cooling The activated Bacillus subtilis ZK-1 strain capable of producing Iturin A and its homologues was inoculated, placed at a temperature of 30° C., and shaken for 18 hours as a seed solution. Use 10 1000mL Erlenmeyer flasks, each containing 300mL medium A (glucose 2.0%, peptone 3.0%, beef extract 0.5%, magnesium sulfate 0.35%, potassium dihydrogen phosphate 0.3%), sterilize at 120°C for 30 minutes, after cooling The bacterial seed liquid is inoculated according to the inoculum amount of 5%-10%, placed at a temperature of 30° C., shaken for 72 hours-120 hours, and fermented at a pH of 6-8.

[0035] After the fermentation, take 2.0L fermented liquid and add water with 200% of the fermented liquid volume to dilute, adjust the pH to 6.0-9.0 with 15% hydrochlor...

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Abstract

The invention belongs to the technical field of biochemical engineering of antibiotics and surfactants, and in particular relates to a method for separating and extracting iturin A (Iturin A), which is produced by microbial fermentation and has surfactant properties and antibacterial activity. The fermented liquid is first treated with flocculants such as aluminum sulfate, polyaluminum chloride, polyferric sulfate, alum, and chitosan, and then iturin A is concentrated and extracted from the fermented liquid. The invention has the advantages of simple process route and low cost, and greatly improves the possibility of industrial and agricultural popularization and application of Iturin A.

Description

[0001] Field [0002] The invention belongs to the technical field of biochemical engineering of antibiotics and surfactants, and in particular relates to a method for separating and extracting iturin A (Iturin A), which is produced by microbial fermentation and has surfactant properties and antibacterial activity. Background technique [0003] Iturin A is a kind of homologue with surfactant properties and antibacterial activity produced by a Bacillus subtilis bacteria isolated from soil by Delcambe and Devignat in 1957. Its structure is as follows: [0004] [0005] Iturin A is one of the earliest and best-known biosurfactants derived from microorganisms. Compared with ordinary surfactants, microbial surfactants not only reduce surface tension, stabilize emulsions and increase foam, but also have non-toxic, biodegradable and stronger surfaces and interfaces that some surfactants do not have. Activity, stability to heat, stability to ionic strength, demulsification, effect...

Claims

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Application Information

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IPC IPC(8): C07K7/64C07K1/30C12P21/04C12R1/125
Inventor 周金燕谭红陈芬钟娟杨杰李志东
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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