Method for producing polymalic acid by high-density fermentation
A polymalic acid and fermenter technology, which is applied in the field of microbial fermentation to achieve the effects of reducing equipment scale, saving power consumption and energy consumption, and improving production intensity
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Embodiment 1
[0024] (1) Activate the Aureobsidium pullulans CGMCC.NO.3336 strain for 6-8 hours, that is, transfer it from the sucrose slant medium to a 500ml baffle bottle with a liquid volume of 50ml and a constant temperature of 25°C. Cultivate at 200r / min for 72h to prepare seed liquid;
[0025] The seed culture medium is composed of: sucrose 140g / L, yeast extract 3g / L, ammonium sulfate 1g / L, succinic acid 2g / L, corn steep liquor 1g / L, K 2 CO 3 0.4g / L, MgSO 4 ·7H 2 O0.1g / L, KH 2 PO 4 1g / L, ZnSO 4 ·7H 2 O0.05g / L, the rest is water, pH7.0.
[0026] (2) Connect the obtained seed liquid to a 5L fermentor at 10% (v / v) for fermentation and culture, according to the OD of the bacteria 600 Value controls pH and dissolved oxygen:
[0027] The first stage of fermentation: when the bacteria OD 600 When it is 0.67, the dissolved oxygen is reduced to 55%, and the stirring is connected in series to maintain the dissolved oxygen at 50%. At the same time, the pH is adjusted to 7.0 with hydrochloric acid and s...
Embodiment 2
[0032] The activation of the strain and the preparation of the seed liquid are the same as in Example 1
[0033] (2) Connect the obtained seed liquid to a 5L fermentor at 10% (v / v) for fermentation and culture, according to the OD of the bacteria 600 Value controls pH and dissolved oxygen:
[0034] The first stage of fermentation: when the bacteria OD 600 When it is 0.65, the dissolved oxygen is reduced to 60%, and the stirring is connected in series to maintain the dissolved oxygen at 55%. At the same time, the pH is adjusted to 6.8 with hydrochloric acid and sodium hydroxide to promote the growth of bacteria;
[0035] The second stage of fermentation: when the bacteria OD 600 When it is 0.75, the dissolved oxygen is reduced to 32%, the temperature is 25°C, and the pH is adjusted to 5.0 with hydrochloric acid and sodium hydroxide, and the fermentation is 104 hours to promote the large-scale synthesis of polymalic acid by the bacteria;
[0036] The concentration of hydrochloric acid is...
Embodiment 3
[0039] The activation of the strain and the preparation of the seed liquid are the same as in Example 1
[0040] (2) Connect the obtained seed liquid to a 5L fermentor at 10% (v / v) for fermentation and culture, according to the OD of the bacteria 600 Value controls pH and dissolved oxygen:
[0041] The first stage of fermentation: when the bacteria OD 600 When it is 0.63, the dissolved oxygen is reduced to 62%, and the stirring is connected in series to maintain the dissolved oxygen at 58%. At the same time, the pH is adjusted to 6.0 with hydrochloric acid and sodium hydroxide to promote the mass growth of bacteria;
[0042] The second stage of fermentation: when the bacteria OD 600 When it is 0.72, the dissolved oxygen is reduced to 35%, the temperature is 25°C, and the pH is adjusted to 4.9 with hydrochloric acid and sodium hydroxide at the same time, and the fermentation is 112 hours to promote the large-scale synthesis of polymalic acid by the bacteria;
[0043] The concentration o...
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