A Novel Method for Simultaneous Biosynthesis of 1,3-Propanediol and Acetoin

A technology for biosynthesis and propylene glycol, applied in the field of bioengineering, can solve problems such as difficulty in refining and separation process

Active Publication Date: 2021-06-11
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the boiling point of 2,3-butanediol is close to that of 1,3-PD, it has caused great difficulties to the subsequent refining and separation process of 1,3-PD

Method used

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  • A Novel Method for Simultaneous Biosynthesis of 1,3-Propanediol and Acetoin
  • A Novel Method for Simultaneous Biosynthesis of 1,3-Propanediol and Acetoin
  • A Novel Method for Simultaneous Biosynthesis of 1,3-Propanediol and Acetoin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. Formed after knocking out the genes encoding PDHC and FdnGHI, the recombinant bacteria synthesized 1,3-PD and AC simultaneously by using glycerol

[0021]M2014574 strain was cultured overnight at 37° C. in LB medium (0.5% yeast extract, 1% tryptone, 1% NaCl, pH 7.0), and the genome was extracted. Using the extracted M2014574 genome as a template, according to the genome sequence (LOCUS: NC_009648) of Klebsiella pneumoniae MGH 78578 registered in NCBI: the upstream of the aceE gene (locus_tag: KPN_00118) and the upstream of the lpdA gene (locus_tag: KPN_00120) Downstream primer design. After the PCR reaction, the target band was recovered from the gel and sequenced. After the sequencing, the gene analysis was performed on the target band, and the similarity with the corresponding fragment gene on MGH 78578 was 99%. Three consecutive genes (7208bp) of aceE-aceF-lpdA in the M2014574 genome were knocked out by means of homologous recombination, and the obtained ...

Embodiment 2

[0026] Example 2, the bacterial strains that knock out the gene encoding PDHC or FdnGHI alone cannot produce the effect of the present invention

[0027] The starting strain M2014574 was used to construct a strain that individually knocked out the gene encoding PDHC or FdnGHI. For specific methods, see Example 1. The strain that individually knocked out the gene encoding PDHC was M2014574△PDHC, and the strain that individually knocked out the gene encoding FdnGHI was M2014574△FdnGHI.

[0028] M2014574, M2014574△PDHC and M2014574△FdnGHI were inoculated into 250ml Erlenmeyer flasks for seed culture, and fermented in a 5L fermenter for 24 hours. The results are shown in Table 2. It can be seen from Table 2 that after knocking out the gene encoding PDHC alone, although 2,3-butanediol was not produced, the cell growth was severely inhibited, 1,3-propanediol was significantly decreased, and AC was not produced. After knocking out the gene encoding FdnGHI alone, the fermentation char...

Embodiment 3

[0031] Example 3. There are three formate dehydrogenases in the genome of Klebsiella pneumoniae, and only by knocking out a specific one of them, that is, the gene encoding FdnGHI, can the effect of the present invention be achieved.

[0032] There are three kinds of formate dehydrogenases (Formate dehydrogenases, FDHs) in K.pneumoniae, respectively: 1) FdhF, that is, FDH-H, encoded by the gene fdhF; 2) FdoGHI, that is, FDH-O, containing three 3) FdnGHI, also known as FDH-N, contains three subunits, encoded by fdnG, fdnH and fdnI, respectively.

[0033] Using the extracted M2014574 genome as a template, primers were designed according to the upstream and downstream of the fdhF gene (locus_tag: KPN_04482) on the genome sequence of Klebsiella pneumoniae MGH78578 registered in NCBI (LOCUS: NC_009648). After the PCR reaction, the target band was recovered from the gel and sequenced. After the sequencing, the gene analysis was performed on the target band, and the similarity with t...

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Abstract

The invention discloses a more effective glycerol utilization method, namely: knocking out the gene encoding formate dehydrogenase and pyruvate dehydrogenase complex in Klebsiella pneumoniae (Klebsiella pneumoniae), and utilizing the obtained recombinant bacteria that simultaneously produce 1,3-propanediol and acetoin. Compared with the existing technology, when using the biotransformation glycerin of the present invention, not only can two high value-added products be produced simultaneously, but also the efficiency of product separation and purification is significantly improved.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, specifically: by knocking out the gene encoding formate dehydrogenase and pyruvate dehydrogenase complex in Klebsiella pneumoniae, simultaneously producing 1,3-propanediol and ethylene glycol marriage. Background technique [0002] With the depletion of fossil fuels, renewable new energy has attracted more and more attention, among which biodiesel has attracted more and more attention because of its advantages of recyclability, degradability, cleanliness and pollution-free. Biodiesel mainly comes from animal and vegetable oils and is prepared by transesterification or thermochemical processes. Therefore, in the production process of biodiesel, a large amount of by-product glycerol will be produced, and the output accounts for about 10% of the total biodiesel production. Therefore, effective methods are needed to make good use of these surplus glycerol. At present, among the methods of ut...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/18C12P7/26C12N1/21C12R1/22
CPCC12N9/0006C12N9/0008C12P7/18C12P7/26C12Y101/05006C12Y102/04001
Inventor 宫衡徐丹凤傅水林
Owner EAST CHINA UNIV OF SCI & TECH
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