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Production method of Sublancin antibacterial peptide

A production method and antimicrobial peptide technology are applied in the production field of Sublancin antimicrobial peptides to achieve the effects of expanding fermentation scale, easy popularization and application, and excellent antibacterial effect

Inactive Publication Date: 2013-01-02
中农颖泰林州生物科园有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The object of the present invention is to provide a method for the fermentative production and purification of a high-purity Sublancin antibacterial peptide, to overcome the deficiencies in the prior art that there is no industrialized production of high-purity Sublancin antimicrobial peptide

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  • Production method of Sublancin antibacterial peptide
  • Production method of Sublancin antibacterial peptide
  • Production method of Sublancin antibacterial peptide

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Embodiment 1

[0043] Fermentation scale: 10m 3 . 1L of fermentation medium contains: 20g of corn flour, 20g of soybean meal, 5g of peptone, 5g of glucose, 5g of ammonium sulfate, and 0.8g of magnesium sulfate. Adjust the pH to 7.5 with sodium hydroxide. The culture solution was sterilized by temperature of 121°C, tank pressure of 0.08Mpa, and holding time of 30min. Bacillus subtilis B. subtilis CVCC20076 was purchased from China Industrial Microorganism Culture Collection Management Center. The inoculum amount of the bacteria was 1×10 6 CFU / mL. Fermentation temperature: 34°C, ventilation: 600m 3 / h, stirring speed: 120r / min, pH7.0. Fermentation culture time: 32h. The membrane pore diameter of ceramic membrane filtration is 20nm to obtain the fermentation broth of Sublacin antimicrobial peptide.

Embodiment 2

[0045] Fermentation scale: 20m 3 . 1L of fermentation medium contains: 50g of corn flour, 50g of soybean meal, 20g of peptone, 20g of glucose, 5g of ammonium sulfate, 5g of yeast powder, and 5g of dipotassium hydrogen phosphate. Adjust the pH to 8.0 with sodium hydroxide. The culture solution is sterilized by a temperature of 121°C, a tank pressure of 0.15Mpa, and a holding time of 30 minutes. The inoculum amount of Bacillus subtilis CVCC20076 was 1×10 7 CFU / mL. Fermentation temperature: 30°C, ventilation: 1000m 3 / h, stirring speed: 160r / min, pH: 6.5. Fermentation culture time: 24h. The membrane pore diameter of ceramic membrane filtration is 10nm to obtain the fermentation broth of Sublacin antimicrobial peptide.

Embodiment 3

[0047] Fermentation scale: 10m 3 . 1L of fermentation medium contains: 10g of corn flour, 10g of soybean meal, 5g of peptone, 2g of sucrose, and 5g of ammonium sulfate. Adjust the pH to 7.0 with sodium hydroxide. The culture solution is sterilized by temperature of 121°C, tank pressure of 0.10Mpa, and holding time of 30 minutes. The inoculum amount of Bacillus subtilis CVCC20076 was 1×10 6 CFU / mL. Fermentation temperature: 37°C, ventilation: 720m 3 / h, stirring speed: 200r / min, pH: 6.0. Fermentation culture time: 28h. The membrane pore diameter of ceramic membrane filtration is 30nm to obtain the fermentation broth of Sublacin antimicrobial peptide.

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Abstract

The invention provides a preparation method of Sublancin antibacterial peptide, which comprises the following steps: (1) inoculating bacillus subtilis into a medium for fermentation, filtering the fermentation liquor; (2) orderly performing centrifugation, cation exchange chromatography, hydrophobic chromatography, size exclusion chromatography, and anion exchange chromatography of the fermentation liquor in step (1), finally performing ultrafiltration concentration and drying to obtain the Sublancin antibacterial peptide. The method of the invention is low in cost, increased in production efficiency, and high in product purity; the prepared Sublancin antibacterial peptide has purity of up to more than 99%; the process is simple, and easy to popularize and apply, and can realize large-scale production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a production method of Sublancin antimicrobial peptide. Background technique [0002] Sublancin antimicrobial peptide is an endogenous polypeptide with antibacterial activity produced by Bacillus subtilis. Synthesized by the ribosome, post-transcriptionally modified and transported by the leader sequence. Its physical property is colorless crystal or white crystalline powder, easily soluble in water. Sublancin contains disulfide bonds, so it has good stability. [0003] Sublancin antimicrobial peptide has a strong inhibitory and killing effect on Gram-positive bacteria, and can be applied to food preservation, anti-infection and inhibition of environmental disinfection. At present, there is no production method of Sublancin antibacterial peptide that can realize industrial production at home and abroad. There are also relatively few reports about the preparation method of Sublanc...

Claims

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Application Information

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IPC IPC(8): C12P21/02C07K1/20C07K1/18C07K1/16C07K14/32C12R1/125
Inventor 郭文江祝发明
Owner 中农颖泰林州生物科园有限公司
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