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Method for producing pediococcusacidilactici by accumulation

A technology of pediocin lactate and inoculum amount, which is applied in fermentation and other directions, can solve the problems of complex operation process, lack of large-scale proliferation of bacteria, long fermentation cycle, etc., and achieve high titer effect

Inactive Publication Date: 2011-07-20
EAST CHINA UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

But the fermentation strategy reported at present mainly adopts the batch fermentation mode, and the minority fed-batch fermentation mode that occurs in recent years, its strategy is relatively single, mainly adopts single pH reduction strategy (Guerra et al, Lett.Appl.Microbiol., 2003) and a single pH cycling strategy (Guerra et al, Process Biochem., 2005), etc., the initial medium used in only a few documents (such as Guerra et al, Enzyme and Microbial Technology, 2007) is generally diluted whey and The mixture of yeast powder, the feed medium is a mixture of lactose (or glucose) and yeast powder, the highest titer is 712AU / ml, and its biggest defect is that the residual sugar (residual glucose in the fermentation broth) concentration is controlled very low (basically 0), so that the bacterium cannot be multiplied in a large amount, and the output of pectin can not be significantly improved, the fermentation cycle is too long (generally exceeding 250 hours), and the operation process is complicated, so it is difficult to realize industrialized production

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  • Method for producing pediococcusacidilactici by accumulation
  • Method for producing pediococcusacidilactici by accumulation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Take 200 μl of Pediococcus lactis preserved in a refrigerator at -20°C and inoculate it into the improved MRS shake flask medium. The medium composition is (g / L): glucose 15.0, peptone 10.0, beef extract 10.0, yeast extract 5.0, (NH 4 ) 2 HC 6 h 5 o 7 2.0, Tween 801.0, H 3 COONa·3H 2 O 5.0, K 2 HPO 4 ·3H 2 O 2.0, MgSO 4 ·7H 2 O 0.58, MnSO 4 ·H 2 O 0.25, distilled water 1000, the pH value was adjusted to 6.5 with 3mol / L hydrochloric acid solution and 3mol / L NaOH solution.

[0021] The Pediococcus LH31 cultivated above was inoculated in a 3.7L fermenter, the inoculum amount was 4wt% based on the volume of the medium, and the initial medium of the fermenter was an improved MRS liquid medium with a pH value of 6.5;

[0022] When the pH value of the fermented liquid drops to 4.2, start feeding. The feeding medium consists of glucose 350g / L and peptone 100g / L. Between 5-10g / L.

[0023] When the pH value of the fermentation broth dropped to 4.5, the pH value of t...

Embodiment 2

[0027] Take 200 μl of Pediococcus lactis preserved in a refrigerator at -20°C and inoculate it into the modified MRS shake flask medium. The medium composition is (g / L): glucose 20.0, peptone 10.0, beef extract 10.0, yeast extract 5.0, (NH 4 ) 2 HC 6 h 5 o 7 2.0, Tween 801.0, H 3 COONa·3H 2 O 5.0, K 2 HPO 4 ·3H 2 O 2.0, MgSO 4 ·7H 2 O 0.58, MnSO 4 ·H 2 O 0.25, distilled water 1000, the pH value was adjusted to 6.5 with 3mol / L hydrochloric acid solution and 3mol / L NaOH solution.

[0028] The Pediococcus LH31 cultivated above was inoculated in a 3.7L fermenter, and the inoculation amount was 4% based on the volume of the medium, and the initial medium of the fermenter was an improved MRS liquid medium, and the pH value was 6.5;

[0029] When the pH value of the fermentation broth dropped to 4.5, feeding was started, and the feeding medium consisted of glucose 300g / L and peptone 40g / L. Feeding adopts a constant-speed feeding method, so that the residual glucose conce...

Embodiment 3

[0034] Take 200 μl of Pediococcus lactis preserved in a refrigerator at -20°C and inoculate it into the modified MRS shake flask medium. The medium composition is (g / L): glucose 18.0, peptone 10.0, beef extract 10.0, yeast extract 5.0, (NH 4 ) 2 HC 6 h 5 o 7 2.0, Tween 801.0, H 3 COONa·3H 2 O 5.0, K 2 HPO 4 ·3H 2 O 2.0, MgSO 4 ·7H 2 O 0.58, MnSO 4 ·H 2 O 0.25, distilled water 1000, the pH value was adjusted to 6.5 with 3mol / L hydrochloric acid solution and 3mol / L NaOH solution.

[0035] The Pediococcus LH31 cultivated above was inoculated in a 3.7L fermenter, and the inoculation amount was 4% based on the volume of the medium, and the initial medium of the fermenter was an improved MRS liquid medium, and the pH value was 6.5;

[0036] When the pH value of the fermentation broth dropped to 4.1, feeding was started, and the feeding medium consisted of glucose 400g / L and peptone 80g / L. Feeding adopts a constant-speed feeding method, so that the residual glucose conce...

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Abstract

The invention relates to a method for producing pediococcusacidilactici by accumulation, comprising the following steps of: inoculating the pediococcusacidilactici in a modified MRS liquid nutrient medium, culturing at 30DEG C for 8 hours, then inoculating the pediococcusacidilactici with the inoculum concentration of 4wt% into a fermentation tank, performing batch fermentation at the first stage of fermentation, performing the second stage of fermentation when the concentration of residual sugar (residual glucose) in the fermentation liquid is below 5-10g / L, and fermenting for 16-30 hours to obtain pediococcusacidilactici with the valence of 18000-24000AU / ml. Compared with the prior art, the method provided by the invention has a short fermentation period and simplicity of operation, and is favorable to quick production of pediococcusacidilactici, and the highest valence of the obtained product is 24000AU / ml.

Description

technical field [0001] The invention relates to a method for producing pediocin lactate, in particular to a method for accumulatively producing pediocin lactate. Background technique [0002] Pediocin is an antibacterial short peptide substance produced by Pediococcus acidilactici belonging to the genus Pediococcus. Studies have shown that nisin is a primary metabolite and a bacteriocin with strong antibacterial activity. It has similar physical and chemical properties and uses to nisin, which has been produced and applied on a large scale. It is resistant to high and low temperature, acid, It is degradable in the human body, so it is safe, non-toxic and convenient to use. Pediocin can inhibit a large number of pathogenic bacteria including Candida albicans, Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Salmonella, Pseudomonas aeruginosa, etc., so it can be used for food preservation and treatment of human diseases. High use value and application prospect. ...

Claims

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Application Information

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IPC IPC(8): C12P21/02
Inventor 孙爱友魏东芝秦鲁敏董玉国徐瑞张星
Owner EAST CHINA UNIV OF SCI & TECH
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