Fermentation method for producing N-acetylneuraminic acid
A technology of acetylneuraminic acid and fermentation method, applied in the field of fermentation for producing N-acetylneuraminic acid, can solve problems such as low yield, limited application scope, high cost, etc., and achieve the effects of increasing yield and improving synthesis rate
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Embodiment 1
[0042] The fermentation method for producing N-acetylneuraminic acid provided in this embodiment includes the following steps:
[0043] 1 Strain activation:
[0044] The frozen N-acetylneuraminic acid-producing strains (preservation number CCTCC No: M2013494) were streaked and inoculated onto solid LB medium and cultured at 37°C.
[0045] 2 Cultivation of primary seeds:
[0046] When a single bacteria grows out, pick a single colony, inoculate it into an Erlenmeyer flask containing 50 mL of liquid LB seed medium, and cultivate it overnight at 37°C at 200 rpm to obtain a first-level seed liquid.
[0047] 3 Cultivation of secondary seeds:
[0048] The first-level seed liquid was inoculated into an Erlenmeyer flask containing 50 mL of liquid seed culture medium at an inoculum of 1%, and incubated at 200 rpm and 37° C. for 6 hours to obtain a second-level seed liquid.
[0049] The liquid LB medium contains: peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L, pH 7.0.
[0050] The s...
Embodiment 2
[0066] The fermentation method for producing N-acetylneuraminic acid provided in this embodiment is basically the same as that of Embodiment 1, except that the feed solution in this embodiment contains 50% glycerol and 2.5% N-acetylglucosamine.
[0067] The results showed that the NeuAc yield was 82.3g / L and the average synthesis rate was 16.46g / L*12h after 60h of fermentation using the fermentation method of this example.
Embodiment 3
[0069] The fermentation method for producing N-acetylneuraminic acid provided in this example is basically the same as that in Example 1, except that the feed solution in this example contains 50% glycerol and 1% polyphosphoric acid.
[0070] The results showed that the NeuAc yield was 76.8 g / L and the average synthesis rate was 15.36 g / L*12h after 60 hours of fermentation using the fermentation method of this embodiment.
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