Preparation method of gamma-linolenic acid
A technology of linolenic acid and seeds, which is applied in the field of preparation of γ-linolenic acid, can solve the problems of low sugar conversion rate, limited application prospect, low fermentation yield, etc., and achieves the effects of low cost, simple operation and low equipment requirements
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Embodiment 1
[0026] 1) Strain activation and shake flask seed culture: under sterile conditions, take 200 μL from the spore glycerol tube stored in a ~80°C ultra-low temperature refrigerator and spread it on the slope of the PDA medium, place it in a 28°C incubator, and adjust according to the growth of the bacteria Cultivate for 7-9 days. Use an inoculation knife to spread 1 cm on the cultured fresh slant 2 The mycelium block accesses the seed culture medium (potato 200g / L; Glucose 20g / L; (NH 4 ) 2 SO 4 10g / L; Tween 800.8mL / 100mL, pH 7.0) in a Erlenmeyer flask, shaker speed 200rpm, 28°C, cultured for 48h.
[0027] 2) Shake flask fermentation culture: the seed solution was inserted into the fermentation medium (glucose 10g / L; sucrose 70g / L; MgSO 4 0.2g / L; KH 2 PO 4 2g / L; Yeast Extract 2.5g / L; Trisodium Citrate 9.6g / L; Citric Acid 2g / L; NH 4 Cl 3g / L; corn steep liquor 5g / L; Tween 800.5mL / 50mL) in a conical flask. At the same time of inoculation, add riboflavin concentrate to the sha...
Embodiment 2
[0032] 1) Strain activation and shake flask seed cultivation are the same as in Example 1;
[0033] 2) Shake flask fermentation culture: the seed solution was inserted into the fermentation medium (glucose 10g / L; sucrose 70g / L; MgSO 4 0.2g / L; KH 2 PO 4 2g / L; Yeast Extract 2.5g / L; Trisodium Citrate 9.6g / L; Citric Acid 2g / L; NH 4 Cl 3g / L; corn steep liquor 5g / L; Tween 800.5mL / 50mL) in a conical flask. At the same time of inoculation, add inositol concentrate to the shake flask medium, so that the concentration of inositol in the medium is 2, 4, 6, 8, 10 mg / L in sequence, and the control addition is 0; the culture temperature is 28°C, and the rotation speed is 200rpm , culture time 168h. At the end of the cultivation, take out the shaker flask, filter it, wash it with deionized water three times, put the bacteria obtained by suction filtration into a weighing bottle (constant weight), and dry it in an oven at 80°C for 24 hours to obtain dried bacteria. Heavy; grind the bacte...
Embodiment 3
[0035] 1) Strain activation and shake flask seed cultivation are the same as in Example 1;
[0036] 2) Shake flask fermentation culture: the seed solution was inserted into the fermentation medium (glucose 10g / L; sucrose 70g / L; MgSO 4 0.2g / L; KH 2 PO 4 2g / L; Yeast Extract 2.5g / L; Trisodium Citrate 9.6g / L; Citric Acid 2g / L; NH 4 Cl 3g / L; corn steep liquor 5g / L; Tween 800.5mL / 50mL) in a conical flask. At the same time of inoculation, add biotin concentrate to the shake flask medium, so that the biotin concentration in the medium is 0.1, 5, 20, 35 mg / L in turn, and the control addition is 0; the culture temperature is 28 ° C, the rotation speed is 200 rpm, and the culture Time 168h. At the end of the cultivation, take out the shaker flask, filter it, wash it with deionized water three times, put the bacteria obtained by suction filtration into a weighing bottle (constant weight), and dry it in an oven at 80°C for 24 hours to obtain dried bacteria. Heavy; grind the bacteria...
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