High-yield quick production method of arachidonic acid
An arachidonic acid, rapid technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the rare problems of phosphorus source research
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Embodiment 1
[0032] Embodiment 1: total phosphorus 0.5g / L in the fermentation medium.
[0033] Strain activation: the Mortierella alpina strain ME-AA01 with the preservation number CCTCC NO: M207067 was inoculated on a PDA slope at 28° C. for 7 days. PDA medium: 200g potatoes, 20g glucose, 20g agar, 1000ml water.
[0034] Seed activation: the PDA slant cultivated for 7 days was washed with sterile water, and inserted into a shaker flask for activation. The shake flask activation medium comprises the following substances in mass-to-volume ratio (% means g / 100mL, the following examples are the same): glucose 3%, yeast extract 0.6%, KH 2 PO 4 0.3%, NaNO 3 0.3%, MgSO 4 ·7H 2 0 0.05%, the rest is water, pH value 7.0, liquid volume: 250ml shake flask is filled with 50ml culture solution, 120rpm, 25°C, activate for 3 days.
[0035] Fermentation culture: the activated seeds are inserted into fermentation shake flasks for fermentation. Inoculum size 10% (v / v), liquid volume: 250 ml shake f...
Embodiment 2
[0039] Embodiment 2: Total phosphorus in the fermentation medium is 0.7g / L.
[0040] Strain activation and seed activation are the same as example 1.
[0041] Fermentation culture: the activated seeds are inserted into fermentation shake flasks for fermentation. Inoculum size 10% (v / v), liquid volume: 250 ml shake flask filled with 50 ml culture solution, temperature 25° C., pH 7.0, rotation speed 120 rpm. The fermentation medium contains the following substances in mass-volume ratio: glucose 8%, yeast extract 0.8%, KH 2 PO 4 0.1%, MgSO 4 ·7H 2 O 0.05%, NaNO 3 0.3%, calcium pantothenate 0.05% and vitamin C 0.02%, the rest is water, pH 7.0.
[0042] After 5 days of fermentation, the wet cells were collected with double-layer filter paper, dried at 60°C, ground, extracted with petroleum ether and ethanol (ratio: 3:1) for 12 hours, and then the petroleum ether and ethanol were distilled off under reduced pressure to obtain oil , dry weight 42.6g / L; fat 21.6g / L; AA13.9g / L...
Embodiment 3
[0046] Embodiment 3: Total phosphorus in the fermentation medium is 0.9g / L.
[0047] Strain activation and seed activation are the same as example 1.
[0048] Fermentation culture: the activated seeds are inserted into fermentation shake flasks for fermentation. Inoculum size 10% (v / v), liquid volume: 250 ml shake flask filled with 50 ml culture solution, temperature 25° C., pH 7.0, rotation speed 120 rpm. The fermentation medium contains the following substances in mass-volume ratio: glucose 8%, yeast extract 0.8%, KH 2 PO 4 0.185%, MgSO 4 ·7H 2 O 0.05%, NaNO 3 0.3%, calcium pantothenate 0.05% and vitamin C 0.02%, the rest is water, pH 7.0.
[0049] After 5.5 days of fermentation, the wet cells were collected with double-layer filter paper, dried at 60°C, ground, extracted with petroleum ether and ethanol (ratio: 3:1) for 12 hours, and then the petroleum ether and ethanol were distilled off under reduced pressure to obtain oil , dry weight 41g / L; fat 21.2g / L; AA12.7g...
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