Method for improving fermentation yield of long-chain dicarboxylic acid
A technology of long-chain dibasic acid and microbial fermentation method is applied in the field of improving long-chain dibasic acid fermentation yield, which can solve the problems of large workload of strain improvement, difficult operation and control, and many uncertain factors, and achieves α- The effect of oxidative decarboxylation and β-oxidation is weak, the acid production rate is fast, and the unit consumption of alkane is reduced
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Embodiment l
[0028] The method for improving the fermentation yield of long-chain dibasic acid comprises the following steps:
[0029] (1) Take an inoculation loop of Candida tropicalis, spread it on the solid slant medium of a large test tube of 20x200mm, and cultivate it at 30°C for 48 hours. The slant medium is solid slant medium (g / L): yeast extract 5, white sugar 10, peptone 5 and agar 15.
[0030] (2) Transfer the bacteria cultured in step (1) into a 500mL Erlenmeyer shaker flask containing 15mL of seed medium, and cultivate for 40 hours at 30°C on a rotary shaker at 180 rpm.
[0031] Seed medium (g / L): White sugar 5, potassium dihydrogen phosphate 8, yeast extract 1.5, corn steep liquor 1.5, urea 1.8, 200# heavy wax oil 30ml / L, prepared with tap water, natural pH.
[0032] (3) In a 500mL Erlenmeyer shaker flask containing 15mL of fermentation medium, add 5mI of the above seed solution, and at the same time add 0.5mmol / L inhibitor chlorpromazine hydrochloride, and make 3 parallel sa...
Embodiment 2
[0036] By the method of embodiment 1, replace chlorpromazine hydrochloride with inhibitor phenobarbital sodium, add when shaking flask inoculation, concentration is 1mmol / L, and fermentation substrate is nC 13 , the amount added is 3ml. The residual hydrocarbon is 0 and the fermentation is over. The dibasic acid is measured by the method of Example 1, and the conversion rate of the fermented alkane is obtained through calculation. ) reached 96.8wt.%, the total acid content (neutralization titration) was 99.1wt.%, and the molar conversion of alkane reached 58.0%.
Embodiment 3
[0038] According to the method of Example 1, replace chlorpromazine hydrochloride with inhibitor acrylic acid, and add when cell culture 28 hours, concentration is 0.05mmol / L, fermentation substrate is nC l4 , the amount added is 3ml. The residual hydrocarbon is 0 and the fermentation ends. After the fermentation, the method of Example 1 was used to measure the dibasic acid, and the conversion rate of the fermented alkane was calculated. As a result, the amount of acid produced was 162.5g / L, and 1.9013g of tetradecanedioic acid was extracted, and the monoacid content (GC) reached 97.5 wt.%, the total acid content (neutralization titration) was 99.4wt.%, and the molar conversion of alkane reached 64%.
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