Pediococcus acidilactici strain and method for producing pediocin by using same

A technology for pediococcus lactis and pediococcus lactis, which is applied in the field of pediocin preparation, and can solve the problems of low yield of pediococcus and low yield of extraction method, etc.

Inactive Publication Date: 2011-07-06
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Another object of the present invention is to provide a method for producing pediocin by culturing the lactic acid Pediococcus (Pediococcus acidilatici) P2 and recovering the method for pediocin from the culture medium and cells, to overcome the wild-type lactic acid in the prior art. Problems of low yield of pediocin from pediococcus and low yield of extraction method

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  • Pediococcus acidilactici strain and method for producing pediocin by using same
  • Pediococcus acidilactici strain and method for producing pediocin by using same
  • Pediococcus acidilactici strain and method for producing pediocin by using same

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Experimental program
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Effect test

Embodiment 1

[0050] The mutant strain P2 was obtained by chemical mutagenesis. Orthogonal test was used to determine the mutagenic conditions of DES as follows: Pediococcus liquid concentration is 10 7 cfu / mL, DES concentration was 0.8%, mutagenesis time was 30min, and mutagenesis temperature was 37℃. In the experiment, 20 mutant strains with increased pediocin yield were obtained. The five strains with the largest increase in yield were selected to continue the LiCl mutation breeding experiment, from which five mutant strains with increased yield were further screened, among which the mutant strain P2 had the highest pediocin titer.

[0051] The specific method of mutagenesis is as follows.

[0052] (1) DES chemical mutagenesis method of strain Pediococcus lactis PA003

[0053] ①The effect of single factor on the lethality of strain Pediococcus lactis PA003

[0054] Concentration of bacterial suspension

[0055] The DES concentration is 0.6%, the mutagenesis time is 20min, the mutagenesis tempera...

Embodiment 2

[0075] Isolation of Mutant Strains of Pediococcus lactis

[0076] Pediococin-producing Pediococcus lactis PA003. The yield of Pediococin was 640AU / mL. The strain was mutagenized with DES, and the high-yielding strain was used as the parent strain and then mutagenized with LiCl. After the mutagenesis test, the P2 mutant strain showed the characteristics of high-producing Pediococin, and the performance of Pediococin production remained stable after the strain was passed through the slope for 20 consecutive generations.

[0077] The P2 mutant strain was inoculated into 10 mL of MRS liquid medium and cultured at 37°C for 16 hours. The antibacterial activity in the culture solution was determined by the above agar diffusion method. The yield of the mutant strain P2 was 19.5 times that of the wild strain PA003. The output is 12,480AU / mL.

Embodiment 3

[0079] Medium and fermentation culture method of mutant strain P2

[0080] A single colony of the mutant strain P2 was inoculated into MRS medium, cultured at 37° C. for 16 hours for activation, and then inoculated into the improved MRS medium provided by the present invention for fermentation culture after activation.

[0081] The fermentation culture method is as follows: the activated mutant strain P2 is inoculated into the modified MRS fermentation medium at a rate of 0.5%, the fermentation temperature is 37℃, the dissolved oxygen in the fermentation broth is maintained at 20%, and the glucose is added at a concentration of 0.025g / L after 7 hours of fermentation For every 5 liters of fermentation broth volume, perform intermittent feeding at a flow rate of 2ml / min, and extract pediocin from the fermentation product after 16 hours of cultivation. Using the above method, the yield of Pediococin is 1.77 times that of normal MRS medium and no added glucose for culture and fermentat...

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Abstract

The invention discloses pediococcus acidilactici P2CGMCC No.4213 and a method for producing pediocin by using the same. The method comprises the following steps of: culturing and activating a pediococcus acidilactici strain P2 and inoculating to an improved Man Rogosa Sharpe (MRS) culture medium for fermentation culture, wherein in the fermentation culture process, the activated pediococcus acidilactici strain P2 is inoculated to the improved MRS fermentation culture medium, the fermentation temperature is between 35 and 37 DEG C, the pH value is 5.5 to 7.5, culture time is 15 to 22 hours, inoculation amount is 0.3 to 0.6 percent, 20 to 25 percent of dissolved oxygen is kept in fermentation liquid, and glucose is fed; and extracting the pediocin from a fermentation product after culturing for 15 to 22 hours. The fermentation is performed through the improved MRS culture medium under the optimized fermentation culture condition, so that the yield of the pediocin is over 22,000AU/ml; and by combining a method for extracting the non-secreted pediocin from cells, the total yield of the pediocin is over 27,500AU/ml, and the pediocin is suitable for large-scale industrial production.

Description

Technical field [0001] The present invention relates to a new strain of Pediococcus acidilatici (Pediococcus acidilatici) and a method for preparing Pediococcus by culturing the strain. Background technique [0002] In order to reduce food spoilage and food-borne diseases caused by microorganisms, humans use methods such as drying, heating, salting, fermentation and chemical preservatives for food preservation. Due to concerns about the side effects of long-term intake of chemical preservatives, consumers prefer not to add preservatives. Preservatives or foods with only natural preservatives, but foods without preservatives are prone to microbial contamination. For example, refrigerated foods contain the psychrophilic pathogens Listeria monocytogenes, Clostridium and Spoilage microorganisms Pseudomonas and Leuconostoc, they cause food safety issues or food shelf life issues. [0003] As the use of chemical preservatives may cause problems, the search for natural metabolites produc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P1/04
Inventor 周志江韩烨李静
Owner TIANJIN UNIV
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