Method for safely producing 1,3-propanediol

A propylene glycol, safe technology, applied in the field of bioengineering, can solve the problems of pathogenicity concerns, insufficient, low pathogenicity correlation, etc.

Inactive Publication Date: 2014-07-02
EAST CHINA UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0008]However, research evidence shows that the frequency of the above-mentioned genes in Klebsiella pneumoniae has little correlation with their pathogenicity, and the above-mentioned genes are used to distinguish various Klebsiella The pathogenicity of P. burgdorferi is not enough (Environ Microbiol, 2004(6), 584-590)
Therefore, simply knocking out the above-mentioned single gene obviously cannot fundamentally relieve people's concerns about its pathogenicity when Klebsiella pneumoniae is used to produce 1,3-PD

Method used

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Embodiment 1

[0021] Example 1 The genes contained in VH1 and VH2 are potential disease-causing genes

[0022] Compare the gene sequences in VH1 and VH2 with the existing virus gene bank. The virus gene bank can be queried in the virus gene database collected and organized by the Institute of Pathogen Biology, Chinese Academy of Medical Sciences, and the Key Laboratory of Pathogen System Biology, Ministry of Health ( http: / / www.mgc.ac.cn / VFs / main.htm). The query results show that the genes contained in the large fragment VH1 and VH2, except fes , wxya , entS In addition, other genes have been confirmed to be disease-causing genes by existing studies, and were originally found in Escherichia coli.

[0023]

Embodiment 2

[0024] Embodiment 2, The reported Klebsiella pneumoniae genome contains large fragments of VH1 and VH2 and is highly homologous

[0025] Table 1: Analysis of VH1 fragments in reported Klebsiella genomes

[0026]

[0027] The genetic data of SEQ ID NO 1 to SEQ ID NO 22 were converted into protein data and compared with the Klebsiella genome data reported on NCBI (http: / / www.ncbi.nlm.nih.gov / ) , the specific compared genomes are: NC_017540, NC_011283, NC_016845, NC_018522, NC_012731, NC_009648.

[0028] Table 2: Analysis of VH2 fragments in reported Klebsiella genomes

[0029]

[0030] After alignment, the genes in SEQ ID NO 1 to SEQ ID NO 22 were located on each genome. Specific genetic data search, comparison, and gene location adopt general methods in molecular biology, and will not be described here. The comparison positioning results are shown in Table 1 and Table 2. As can be seen from Table 1 and Table 2, the genomes of each Klebsiella pneumoniae contain VH1 a...

Embodiment 3

[0032] Embodiment 3, VH1 and VH2 gene fragments also exist in 1,3-PD Klebsiella pneumoniae

[0033] The 1,3-PD-producing strain uses ATCC49790. The upstream and downstream primers of VH1 are derived from the gene fepA (SEQ ID NO 1) upstream and gene entA (SEQ ID NO 13). The upstream and downstream primers of VH2 are derived from the gene fimB (SEQ ID NO 14) upstream and gene f (SEQ ID NO 22).

[0034] ATCC49790 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 ATCC49790 genome as a template, the designed VH1 and VH2 upstream and downstream primers were used for PCR reaction. After the PCR reaction, the target band was recovered and sequenced, and the gene analysis was performed on the target band after sequencing.

[0035] The analysis results are shown in Table 3. As can be seen from Table 3, the genome of ATCC49790 contains VH1 (17678bp) and VH2 (8761bp) fragments, and ...

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Abstract

The invention discloses a method for safely producing 1,3-propanediol, namely the safety of 1,3-propanediol fermentation is improved by knocking-out continuous emerging klenow fragment genes in a virus gene cluster VH1 and / or VH2 in klebsiella peneumoniae. Compared with an existing method for producing 1,3-propanediol with converted glycerol, the method disclosed by the invention has the advantages that latent pathogenic genes in K. peneumoniae are knocked out in klenow fragment and the biosecurity of klebsiella peneumoniae is improved.

Description

[0001] technical field [0002] The invention belongs to the technical field of bioengineering, specifically, knocking out the large fragment gene causing potential pathogenicity in Klebsiella pneumoniae, thereby improving the safety of 1,3-propanediol fermentation, and at the same time having no effect on production performance Influence. [0003] Background technique [0004] 1, 3-propanediol (1, 3-porpanediol, referred to as 1, 3-PD) is an important chemical raw material, which can be used as a solvent, antifreeze or protective agent, fine chemical raw material and a new type of polyester - poly Monomer of propylene glycol phthalate (PTT) and polyurethane. PTT has been proved to be a new type of polyester material with excellent performance. The existing 1, 3-PD production methods include chemical synthesis and biosynthesis. At present, the synthesis of 1, 3-PD in the world is mainly carried out by chemical methods. However, compared with chemical synthesis, biologic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/18C12R1/22
Inventor 宫衡高黎荣蒋潇付水林
Owner EAST CHINA UNIV OF SCI & TECH
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