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A kind of method and fermentation broth for improving fermentation to produce polysialic acid

A technology of polysialic acid and fermentation broth, applied in the field of fermentation engineering, can solve the problems of low product concentration, long fermentation period, low production efficiency, etc., and achieve the effects of high cumulative content, simple process and high production efficiency

Active Publication Date: 2021-01-26
武汉中科光谷绿色生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Continuous fermentation can greatly increase the production intensity, but it is easy to infect bacteria, strain degradation, large equipment investment and low product concentration.
Batch fermentation consumes a lot of manpower and material resources. Each batch of fermentation needs to be loaded, sterilized, inoculated, discharged, and cleaned. The process is cumbersome, the fermentation cycle is long, and the production efficiency is low.
Although fed-batch fermentation can supplement the deficiency of nutrients or precursors through feeding, the product synthesis will inevitably be blocked due to the continuous accumulation of harmful metabolites

Method used

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  • A kind of method and fermentation broth for improving fermentation to produce polysialic acid
  • A kind of method and fermentation broth for improving fermentation to produce polysialic acid

Examples

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preparation example Construction

[0037] In the present embodiment, the preparation method of seed culture solution comprises the following processes:

[0038] (1) Activation of strains: Inoculate frozen polysialic acid-producing strains on solid LB medium and culture at 35-40°C. Among them, the solid LB medium contains: peptone 8-12g / L, yeast extract powder 4-6g / L, sodium chloride 8-12g / L, agar 15-25g / L, pH7.0±0.2.

[0039] (2) Cultivation of first-class seeds: after a single colony grows, pick a single colony, inoculate it into a liquid LB medium, and cultivate it overnight at 150-250 rpm and 35-40° C. to obtain a first-class seed liquid. Among them, the liquid LB medium contains: peptone 8-12g / L, yeast extract powder 4-6g / L, sodium chloride 8-12g / L, pH7.0±0.2.

[0040] (3) Cultivation of secondary seeds: the primary seed liquid is received in the liquid LB medium according to the inoculum size of 0.8%-1.2%, and cultivated at 150-250rpm, 35-40°C for 5-7h to obtain the secondary seed liquid , which is the s...

Embodiment 1

[0047] The present embodiment provides a fermented liquid, which is prepared according to the following preparation method:

[0048] 1. Strain activation: Streak inoculation of frozen polysialic acid-producing Escherichia coli (preservation number CCTCC NO: M 2018103) on solid LB medium, and culture at 37°C. Among them, the solid LB medium contains: peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L, agar 20g / L, pH7.0.

[0049] 2. Cultivation of first-class seeds: After a single colony grows, pick a single colony, inoculate it into a conical flask containing 50 mL of liquid LB medium, and cultivate it overnight at 200 rpm at 37° C. to obtain a first-class seed liquid. Among them, the liquid LB medium contains: peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L, pH7.0.

[0050] 3. Cultivation of secondary seeds: The primary seed solution was transferred to a conical flask containing 50 mL of liquid seed medium at an inoculation amount of 1%, and cultu...

Embodiment 2

[0058] This embodiment provides a fermented liquid, the preparation method of this fermented liquid is roughly the same as the preparation method in Example 1, the difference is:

[0059] Every 60 hours, 90% of the volume of the fermentation broth was drained, and the same volume of fresh liquid culture medium without glycerol was added.

[0060] Get the fermented liquid discharged from each batch, and detect its OD600 and polysialic acid content.

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Abstract

The invention provides a method for increasing polysialic acid produced by fermentation and fermentation liquor and relates to the technical field of fermentation engineering. The method for increasing polysialic acid produced by fermentation in the embodiment comprises steps as follows: a liquid fermentation medium is inoculated with seed culture fluid capable of producing polysialic acid strainsfor fermentation, a volume ratio of the liquid fermentation medium to the seed culture fluid is 25-35:1, pH keeps 6-7 in the fermentation process, and after a carbon source in the liquid fermentationmedium is used up, the carbon source is supplemented at a rate of 2-3 g / (L.h); 85%-95% by volume of fermentation liquor is discharged every 48-60 h, and the same volume of fresh liquid medium containing no carbon source is supplemented. The process of the method is simple, and production efficiency is high. The fermentation liquor in the embodiment is prepared with the method for increasing polysialic acid produced by fermentation, and the accumulative content of polysialic acid is high.

Description

technical field [0001] The invention relates to the technical field of fermentation engineering, and in particular to a method and fermentation liquid for improving polysialic acid produced by fermentation. Background technique [0002] On May 31, 2017, the National Health and Family Planning Commission reviewed and approved the use of 10 new food ingredients including N-acetylneuraminic acid (Neu5Ac). Neu5Ac is the first contact site for cell information transmission, and its molecular structure is diverse, so Neu5Ac is involved in multiple physiological processes such as cell recognition, signal transduction, tumorigenesis, and fertilization. In addition, Neu5Ac can regulate the anti-inflammatory activity of IgG, enhance infant immunity, affect the integrity, permeability and activity of nerve cells, and promote the function of infant brain development, so N-acetylneuraminic acid and its production have caused more Follow and research. [0003] The current production met...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/26C12N1/20C12R1/19
CPCC12N1/20C12P19/26
Inventor 吴金勇陈祥松李翔宇刘洋姚建铭
Owner 武汉中科光谷绿色生物技术有限公司