Method for co-production of 1,3-propanediol and d-1,2,4-butanetriol
A technology of D-1 and propylene glycol, applied in the fields of fermentation and biology, can solve the problems of high price of xylose, low conversion rate, limitation of xylose, etc.
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Embodiment 1
[0049] Example 1: Construction of recombinant Klebsiella pneumoniae
[0050] Specific steps are as follows:
[0051] Synthesize the gene yjhG (SEQ ID No. 6) encoding xylonate dehydratase derived from Escherichia coli; combine the synthesized gene kivD encoding 2-ketoisovalerate decarboxylase with the recombinant plasmid pEtac-kivD-tac -xdh (recorded in the literature "Jing P Y, Cao X, Lu X Y, etc. Modification of an engineered Escherichia coli by a combined strategy of deleting branch pathway, fine-tuning xylose isomerase expression, and substituting decarboxylase to improve1,2,4-butanetriol production[J]. Journal of Bioscience and Bioengineering, 2018, 126(5): 547-552.", the nucleotide sequence of the tac promoter is shown in SEQ ID No. 7) by restriction endonuclease Xba I After digestion with Nco I, ligation was performed to obtain a ligated product; the ligated product was transformed into Escherichiacoli BL21 to obtain a transformed product; the transformed product was co...
Embodiment 2
[0052] Example 2: Co-production of 1,3-propanediol and D-1,2,4-butanetriol
[0053] Specific steps are as follows:
[0054] With Klebsiella pneumoniae (Klebsiella Pneumoniae) JI22914 as a control, the single colony of the recombinant Klebsiella pneumoniae JI22914 / pEtac-yjhG-tac-kivD-tac-xdh obtained in Example 1 was streaked in LB solid medium medium, incubate at 37°C for 12-14h to form a single colony; pick a single colony and inoculate it into LB liquid medium, cultivate at 37°C and 100rpm for 12h to obtain first-class seed liquid; inoculate according to 1% (v / v) Transfer the first-class seed solution to a new LB liquid medium, and cultivate at 37°C and 100 rpm for 8 h to obtain the second-class seed solution; transfer the second-class seed solution to a In the fermentation medium, shake culture at 37° C. and 150 rpm for 48 hours to obtain a fermentation broth.
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