Method for efficiently producing 1, 3-propylene glycol by reducing by-product acetic acid
A propylene glycol, high-efficiency technology, applied in the field of bioengineering, can solve problems such as no detailed research and no reports, and achieve the effect of fast product synthesis, less by-products, and high conversion rate
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[0022] Example 1. Knocking out the poxB gene alone severely reduces the growth of the bacteria, which is not conducive to the synthesis of 1,3-PD
[0023] The M2014574 strain was cultured in LB medium (0.5% yeast extract, 1% tryptone, 1% NaCl, pH 7.0) at 37°C overnight to extract the genome. Using the extracted M2014574 genome as a template, design primers based on the poxB gene (locus_tag: KPN_00904) on the genome sequence of Klebsiella pneumoniae MGH78578 registered by NCBI (LOCUS: NC_009648), and the target band was recovered by the gel after the PCR reaction. And sequenced. After sequencing, the target band was genetically analyzed, and the similarity with the poxB gene on MGH78578 was 100%. Homologous recombination was used to knock out the poxB gene (1278bp) of the M2014574 genome, and the resulting recombinant strain was M2014574△poxB.
[0024] M2014574 and M2014574△pta were respectively inoculated into a 250ml Erlenmeyer flask and cultured at 37°C for 12 hours anaerobic an...
Example Embodiment
[0028] Example 2. Knockout of pta gene alone has almost no effect on bacterial growth and 1,3-PD synthesis
[0029] The M2014574 strain was cultured in LB medium (0.5% yeast extract, 1% tryptone, 1% NaCl, pH 7.0) at 37°C overnight to extract the genome. Using the extracted M2014574 genome as a template, design primers based on the pta gene (locus_tag: KPN_02688) of the Klebsiella pneumoniae MGH78578 genomic sequence (LOCUS: NC_009648) registered in NCBI, and the target band was recovered by the gel after the PCR reaction. And sequenced. After sequencing, the target band was genetically analyzed, and the similarity with the poxB gene on MGH78578 was 100%. Homologous recombination was used to knock out the poxB gene (2133bp) of the M2014574 genome, and the resulting recombinant strain was M2014574△pta.
[0030] M2014574 and M2014574△pta were respectively inoculated into 250ml Erlenmeyer flasks and cultured at 37°C for 12 hours anaerobic and aerobic. The medium was seed medium with 4...
Example Embodiment
[0034] Example 3. Knockout of poxB and pta genes at the same time did not greatly inhibit the bacteria, but the synthesis of 1,3-PD increased
[0035] Klebsiella pneumoniae M2014574 was simultaneously knocked out according to the methods of Examples 1 and 2, and the recombinant strain obtained was M2014574△poxB-pta.
[0036] M2014574 and M2014574△poxB-pta were respectively inoculated into 250ml Erlenmeyer flasks and cultured at 37°C for 12 hours anaerobic and aerobic. The medium was seed medium with 40g / L glycerol. During aerobic culture, the Erlenmeyer flask was sealed with 8 layers of gauze; during anaerobic culture conditions, the air in the flask before inoculation was replaced with nitrogen and the Erlenmeyer flask was sealed with a rubber stopper.
[0037] After culturing for 12 hours, the bacterial cell concentration, 1,3-PD, and acetic acid concentration in the fermentation broth were measured. The results are shown in Table 3. It can be seen from Table 2 that after both pox...
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