Long chain normal dibasic acid production method
A long-chain dibasic acid and production method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, separation/purification of carboxylic acid compounds, etc., can solve the problem of increasing production costs, high prices, and downstream product applications Field limitations and other issues to achieve the effect of expanding applications
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Embodiment 1
[0022] Embodiment 1: the preparation of seed solution
[0023] 1) Take the glycerol tube strain of Candida tropicalis (CCTCC NO: M203052) and inoculate it into 500 mL seeds filled with 200 mL liquid medium (10 g / kg peptone, 5 g / kg yeast extract, 10 g / kg glucose, natural pH) Cultivate in the bottle at 35°C, 300rpm shaker for 10-15 hours;
[0024] 2) Take the above-mentioned shaking flask seeds, insert them into a 10L fermenter equipped with 5L seed medium (10g / kg peptone, 10g / kg yeast extract, 10g / kg glucose, natural pH), and cultivate them at 35°C for 24 hours to produce Get the primary seed solution for later use.
[0025] 3) With 16M 3 Medium 20M 3 In the fermenter, the above-mentioned first-level seed liquid is inserted, and the second-level seed tank cultivation is started. The fermentation medium formula is the same level as the seed tank. Cultivate at 29°C for 16 hours to prepare secondary seed solution for future use.
Embodiment 2
[0026] Example 2 Oleic acid fermentation
[0027] In a 30L fermenter equipped with 15L medium, respectively insert 1.5L of the primary seed solution of Example 1 to start fermentation. The fermentation medium of the fermenter is: potassium dihydrogen phosphate 10g / kg, magnesium sulfate 0.5g / kg, urea 5.0g / kg, corn steep liquor 10g / kg, glucose 40g / kg, pH 5.8, sterilized at 121°C for 20 minutes . Dissolution of oleic acid, feed sugar and alkali. Cultivate at 35°C, rotation speed 500rpm, ventilation rate 7.5vvm, and tank pressure 0.05Mpa. After 3 hours of fermentation, start adding glucose at a rate of 100g / hr. After 48 hours, reduce the speed of sugar flow to 70g / hr. After 96 hours Reduce the sugar flow rate to 40g / hr, and stop adding sugar 5 hours before the end of fermentation; 3 hours after adding glucose, start to flow oleic acid at a rate of 30g / hr, and stop oleic acid flow about 20 hours before the end of fermentation. From inoculation to the end of fermentation, the tot...
Embodiment 3
[0028] Embodiment 3 Undecyl n-alkanes (DC 11 ) fermentation
[0029] installed in 100M 3 200M of medium 3 In the fermenter, the primary seed liquid is inserted to start fermentation. The composition of the fermentation medium is: glucose 10g / L, KH 2 PO 4 15g / L, yeast extract 10g / L, corn steep liquor 5g / L, urea 4.5g / L, NaCl 1g / L, KNO3 7g / L, natural pH, sterilized at 121℃. Dissociation of alkanes and feed sugars. Cultivate at 29°C under the conditions of ventilation rate 0.5vvm and tank pressure 1.0Mpa. 20 hours before fermentation, the pH was natural, and the growth of bacteria was the main factor. When the growth optical density (OD 600 ) greater than 0.6, start to add C 11 Alkanes, to control the C in the fermentation broth 11 The alkane concentration is maintained at about 5% (V:V), and the pH is adjusted to 7.0 at the same time. Within 48 hours, the pH is adjusted to 7.0 with NaOH solution every 4 hours. For 48-72 hours, the pH is adjusted to 7.5 with NaOH soluti...
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