A method for reducing non-target carbon chain length dibasic acid impurities in the production of dibasic acids
A carbon chain length, dibasic acid technology, applied in the field of dibasic acid impurities, can solve problems such as the accumulation of metabolic by-products, affecting the quality of downstream products, and the production of dibasic acid impurities
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0072] Example 1 Construction of long-chain dibasic acid-producing strains
[0073] 1. Cloning of FAS2 gene
[0074] (1) Total RNA extraction and transcriptome sequencing
[0075]A single colony of Candida tropicalis strain CAT N145 was picked and inoculated into a 10 ml test tube containing 1 ml of YPD medium (containing 100 mg / L hygromycin B), and the bacterial liquid was collected after enrichment by conventional methods. RNA was extracted using TRNzol universalReagen (Tiangen) kit, supplemented by liquid nitrogen trituration to disrupt cells. Transcriptome sequencing was performed using the Miseq (Illumina) platform, and paired-end sequencing was used to obtain 22M reads with a length of 2 x 251bp. After removing the linker and filtering low-quality bases and reads with CutAadpt, the measured Reads were assembled with Trinity software (http: / / trinityrnaseq.sf.net) to obtain Unigene, and functionally annotated with NCBI's Non-Redundant protein database.
[0076] (2) Bioi...
Embodiment 2
[0110] Embodiment 2 Recombinant strain 662 fermentation produces twelve carbon long carbon chain dibasic acid
[0111] A single colony of strain 662 was picked and inoculated into a 10 ml test tube containing 1 ml of YPD medium (containing 100 mg / L hygromycin B) at a temperature of 31 °C and a shaker speed of 220 rpm for 24 hours. Get the above-mentioned bacterial liquid and insert it into a 500mL shake flask containing 25ml of seed medium, the inoculum size is 7%, the shaker rotation speed is 220rpm, and the culture temperature is 31°C and cultivated to OD. 620 when reaching 0.8. The seed liquid was inserted into a 500 mL shake flask containing 15 mL of fermentation medium, the inoculation amount was 20%, and the substrate in the fermentation medium was 250 mL / L of n-dodecane. The culture was carried out at 31° C. and 250 rpm in a shaker until the end of the fermentation, and the pH was controlled to be 7.5 to 7.6 during the culture. The strain CAT N145 was used as the cont...
Embodiment 3
[0116] Example 3 Fermentation of strain 662 to produce ten-carbon long-chain dibasic acids
[0117] The fermentation method was the same as in Example 2, except that the substrate in the fermentation medium was 250 mL / L n-decane. The strain CAT N145 was used as a control group: the culture and fermentation methods were the same as those of strain 662 except that the medium did not contain hygromycin B.
[0118] Get 0.5g above-mentioned fermentation broth sample respectively, carry out GC detection, calculate the content of decanedicarboxylic acid and the impurity mass ratio of dodecanedioic acid and tetradecanedicarboxylic acid (the impurity mass ratio is that the impurity accounts for the target length in the fermentation broth. The mass percentage of chain dibasic acid), the results are shown in Table 2 below.
[0119] Table 2
[0120] strain CAT N145 662 Decadic acid yield (mg / g) 109.6 112.5 Dodecanedioic acid (%) 1.21 0.60 Tetradecanoic ac...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More