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A method for producing sialic acid through fermentation

A technology of sialic acid and fermentation broth, applied in the field of bioengineering, can solve the problems of no further hydrolysis, no report of sialic acid content, etc.

Active Publication Date: 2017-04-05
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The relevant literatures that have been reported so far are almost all the detected polysialic acid content, the maximum yield is 6.92g / L, there is no further hydrolysis, or the sialic acid is directly purified after hydrolysis, and the content of sialic acid is not reported

Method used

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  • A method for producing sialic acid through fermentation
  • A method for producing sialic acid through fermentation
  • A method for producing sialic acid through fermentation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The optimization of the carbon and nitrogen source of embodiment 1 fermentation medium

[0049] 1. Determination of carbon source content in fermentation medium

[0050] Experiment with three kinds of content (30, 40, 50g / L) of carbon source sorbitol with single factor experiment, determine suitable carbon source content, name it as A1, A2, A3 respectively, other components and content in the fermentation medium are : Dipotassium hydrogen phosphate 2.5g / L, ammonium sulfate 5g / L, yeast extract 1.2g / L, magnesium sulfate 0.9g / L, copper sulfate 0.002g / L, pH8.0 (sterilized at 121°C for 15min).

[0051] 500mL three-neck flask culture: the cultured seed solution was inserted into a 500mL three-neck flask with 300mL fermentation medium according to the inoculum amount of 4% (v / v) for cultivation, the shaker speed was 250r / min, the temperature was 37°C, and cultured for 40h. Sampling at regular intervals, measuring the mass concentration of sorbitol and the bacterium absorbance...

Embodiment 2

[0062] The influence of embodiment 2 hydrogen peroxide stress on thalline fermentation ability

[0063] 1. The effect of the addition time and amount of hydrogen peroxide on the yield of sialic acid in the final product

[0064] Carry out the following single factor experiment respectively, when fermenting to 5h, add the H of 1, 2, 4, 8mmoL / L respectively 2 o 2 , respectively named as C1, C2, C3, and C4; when the fermentation reached 10h, 2, 4, 8, and 16mmoL / L of H 2 o 2 , respectively named D1, D2, D3, D4; when fermented to 15h, add 4, 8, 16, 32mmoL / L of H 2 o 2 , respectively named E1, E2, E3, E4.

[0065] 500mL three-necked flask culture: put the cultured seed solution into a 500mL three-necked flask with 300mL fermentation medium according to the inoculum amount of 4% (v / v) for cultivation, the shaker speed is 250r / min, the temperature is 37°C, and the pH is 7.6 ~8.0, cultivated for 40 hours, measured the content of sialic acid at the end of fermentation, and made a ...

Embodiment 3

[0077] The influence of embodiment 3 rotating speed on thalline fermentation ability

[0078] During the fermentation process, different rotational speeds were set to 300, 400, 500, and 600 r / min, respectively, and single-factor experiments were carried out.

[0079] 500mL three-necked flask culture: put the cultivated seed solution into 300mL fermentation medium according to the inoculum size 4% (v / v), rotate at 250r / min, temperature 37°C, pH7.6~8.0, cultivate for 40h, Sampling, measuring the content of sialic acid in the fermentation broth and the dry weight of bacteria, by analyzing the influence of different stirring speeds on the specific growth rate of bacteria cells and the specific synthesis rate of sialic acid, determine the optimal rotating speed of segmental stirring and the effect of segmental stirring. point in time.

[0080]Scale-up culture: 3.7L fermenter with 2L liquid volume, 4% inoculum, 37°C fermentation temperature, 250r / min stirring speed, 8.0 pH, 1 (V / V·...

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Abstract

The invention discloses a fermentation production method of sialic acid and belongs to the technical field of bioengineering. The method comprises the following steps: adding a seed liquid into a fermentation tank containing a sterilizing fermentation culture medium; continuously fermenting to produce fermentation liquid containing polysialic acid in a way of replenishing feeding liquid and a hydrogen dioxide solution; then carrying out centrifugal separation, ethanol precipitation and filtration to obtain refined polysialic acid; then, carrying out acidolysis and crystallization on the polysialic acid; and finally, washing and freeze-drying to obtain sialic acid powder. The output of the polysialic acid produced by the method provided by the invention can reach 12.96g / L, the already reported maximum content of the polysialic acid is improved by 87.28%, the hydrolysis rate of polysialic acid is 95%, the content of purified sialic acid reaches 10.01g / L and the purity reaches 94.80%. The method is suitable for industrial promotion and application.

Description

technical field [0001] The invention relates to a method for fermenting and producing sialic acid, belonging to the technical field of bioengineering. [0002] technical background [0003] Sialic acid (Sialic acids) is a family of neuraminic acid derivatives, the most common being N-acetylneuraminic acid (NANA), usually in the form of oligosaccharides, glycolipids or glycoproteins. The main food source of sialic acid is breast milk, but sialic acid is also found in milk, eggs, and cheese. In recent years, the application of sialic acid and its derivatives in food, health care products and medicine has increasingly broad development prospects, especially in infant formula milk powder. Studies have found that exogenous sialic acid can be supplemented by diet to Increase the content of sialic acid in the brain. This also indicates that adding sialic acid in infant milk powder, especially infant milk powder for premature infants, will effectively promote the development of the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/26
Inventor 刘丽波谢凤英李春迟涛刘宁王敏梁婉婷祁昕
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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