Novel biological fermentation method for normal long-chain tridecanedioic acid
A thirteen carbon dibasic acid and biological method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problem that the product purity cannot reach the long-chain dibasic acid ester, the fermentation level is not high, Inconvenience and other issues
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Embodiment 1
[0061] (1) Take an inoculation loop of Candida tropicalis C1301 cells, spread it on the wort solid slope of a 15×180 large test tube, and cultivate it at 28° C. for 2 days.
[0062] (2), get one of the above strains, insert it into a 250ml Erlenmeyer flask with 30ml liquid seed culture medium and cultivate it for 40 hours at 28-30°C on a rotary shaker at 220 rpm. Liquid seed medium contains KH 2 PO4 8g / l, yeast extract 5g / l, corn steep liquor 3g / l, sucrose 30g / l, urea 3g / l, tap water, pH 5.0.
[0063] (3) In a 500ml Erlenmeyer flask containing 15ml of fermentation medium, insert 3.5ml of the above-mentioned cultivated seed solution, ferment for 4 days at 28-30°C, 220 rpm on a rotary shaker, and use 6N every 24 hours Adjust the pH to 7.5-8.0 once with NaOH. The fermentation medium mixture contains KH 2 PO 4 10g / l, sucrose 20 / l, yeast extract 3g / l, corn steep liquor 3.5g / l, urea 1.5g / l, and nC13200ml / l, tap water configuration, PH7.2. Sterilize at 110°C for 30 minutes.
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Embodiment 2
[0066] (1) Seed medium and culture method and fermentation medium are the same as example 1.
[0067] (2) Cultivate 3000mL for two days, OD(×30, 620nm) is 0.81pH 3.8, strong, no miscellaneous bacteria seed solution into the first-level seed tank with 700L seed medium, sterilized at 121°C for 40 minutes , 29°C 350 rpm, tank pressure 0.8kg / cm 2 , ventilation rate 1:0.8, cultured for 36 hours, as the seeds of the secondary seed mother.
[0068] (3) Insert the 700L seed solution cultivated in (2) into a 6.5m 3 Seed medium, 10m after sterilized at 121°C for 40 minutes 3 In the secondary seed tank, 30°C, 200 rpm, tank pressure 1kg / cm 2 , ventilation rate 1:0.7, cultivated for 40 to 48 hours, as fermented seeds.
[0069] (4) Connect the strong and bacteria-free seed solution cultivated in (3) into a 33m 3 Fermentation medium, 50m of sterilized at 121°C for 40 minutes 3 In a fermenter, 30°C, 200 rpm, tank pressure 1kg / cm 2 , ventilation volume 1:0.5, at the beginning, nC 13 4...
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