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Method for promoting Escherichia coli fermentation to produce polysialic acid

A technology of Escherichia coli and polysialic acid, applied in the field of fermentation engineering, can solve the problems of limiting the application of N-acetylneuraminic acid, low synthesis efficiency and carbon source conversion efficiency, and high fermentation cost

Active Publication Date: 2020-07-07
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

However, at present, the fermentation level, synthesis efficiency and carbon source conversion efficiency of polysialic acid are low, and the yield of polysialic acid PSA is only 5-6 g / L when fermented for 30-40 hours, which leads to high fermentation cost and limits N- Application of acetylneuraminic acid

Method used

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  • Method for promoting Escherichia coli fermentation to produce polysialic acid
  • Method for promoting Escherichia coli fermentation to produce polysialic acid
  • Method for promoting Escherichia coli fermentation to produce polysialic acid

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Embodiment 1

[0036] The embodiment of the present invention provides a method of using Escherichia coli to ferment and produce polysialic acid, which mainly includes the following steps:

[0037] S11. Streak inoculation of frozen Escherichia coli (preservation number CCTCC NO: M 2018103) on solid LB medium and culture at 37°C to obtain a single colony; the Escherichia coli used was obtained on March 6, 2018 It has been submitted to the China Center for Type Culture Collection (CCTCC) in Luojia Mountain, Bayi Road, Wuchang District, Wuhan City, Hubei Province for preservation; wherein, the solid LB medium contains the following components: peptone 10g / L, yeast extract powder 5g / L , sodium chloride 10g / L, agar 20g / L; the pH of the solid LB medium is 7.0.

[0038] S12. Pick a single colony, inoculate it into an Erlenmeyer flask containing 50 mL of liquid LB seed medium, and cultivate it overnight at a stirring rate of 200 rpm at 37° C. to obtain a first-grade seed liquid; wherein, the liquid ...

Embodiment 2

[0044] The embodiment of the present invention provides a method of using Escherichia coli to ferment and produce polysialic acid, which mainly includes the following steps:

[0045] S21. Streak inoculation of frozen Escherichia coli (preservation number CCTCC NO: M 2018103) on solid LB medium and culture at 37°C to obtain a single colony; wherein, the solid LB medium contains the following components: peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L, agar 20g / L; the pH of the solid LB medium is 7.0.

[0046] S22. Pick a single colony, inoculate it into an Erlenmeyer flask containing 50 mL of liquid LB seed medium, and cultivate it overnight at a stirring rate of 200 rpm at 37° C. to obtain a first-grade seed liquid; wherein, the liquid LB seed medium contains the following components : Peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L; the pH of the liquid LB seed medium is 7.0.

[0047] S23. The primary seed solution was transferred to a conical...

Embodiment 3

[0052] The embodiment of the present invention provides a method of using Escherichia coli to ferment and produce polysialic acid, which mainly includes the following steps:

[0053] S31. Streak inoculation of frozen Escherichia coli (preservation number CCTCC NO: M 2018103) onto solid LB medium and culture at 37°C to obtain a single colony; wherein, the solid LB medium contains the following components: peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L, agar 20g / L; the pH of the solid LB medium is 7.0.

[0054] S32. Pick a single colony, inoculate it into an Erlenmeyer flask containing 50 mL of liquid LB seed medium, and cultivate it overnight at a stirring rate of 200 rpm at 37° C. to obtain a first-grade seed liquid; wherein, the liquid LB seed medium contains the following components : Peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L; the pH of the liquid LB seed medium is 7.0.

[0055] S33. Transfer the primary seed liquid into a conical flas...

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Abstract

The invention relates to a method for promoting fermentation of Escherichia coli to produce polysialic acid, and relates to the fermentation engineering field. The method for promoting fermentation ofEscherichia coli to produce polysialic acid comprises the following steps: inoculating Escherichia coli into fermentation medium for culture and fermentation, and separating to obtain polysialic acid; when fermenting, controlling the fermentation of the temperature between 0-48 hours to be 40-42 DEG C, the fermentation temperature after 48 hours to be 36 DEG C to 38 DEG C. The method for promoting the fermentation of Escherichia coli to produce polysialic acid can effectively improve the fermentation level, synthesis efficiency and carbon source conversion efficiency of polysialic acid.

Description

technical field [0001] The invention relates to the field of fermentation engineering, and in particular to a method for promoting the fermentation and production of polysialic acid by Escherichia coli. Background technique [0002] N-acetylneuraminic acid (N-acetylneuraminic acid) 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 , Neu5Ac can also regulate the anti-inflammatory activity of IgG, enhance infant immunity, affect the integrity, permeability and activity of nerve cells, and promote the development of infant brain, so the production of N-acetylneuraminic acid has attracted more attention and attention. Research. [0003] The current production methods of N-acetylneuraminic acid include: natural product extraction, chemical synthesis, bio-enzymatic conversion, direct fer...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/26C12R1/19
CPCC12P19/26
Inventor 吴金勇陈祥松李翔宇朱薇薇姚建铭
Owner 武汉中科光谷绿色生物技术有限公司
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