Klebsiella for strengthening expression of citT gene and application of klebsiella in production of 1,3-propylene glycol

A Klebsiella, propylene glycol technology, applied in the direction of microorganism-based methods, bacteria, microorganisms, etc., can solve the problem of no research report on the citT gene of Klebsiella

Active Publication Date: 2017-11-07
ZHANGJIAGANG GLORY CHEM IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The genome annotation results of Klebsiella show that many Klebsiella have the citT gene, but there is no research report on the function of the Klebsiella citT gene

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The present embodiment provides a construction method for enhanced expression of citT genetically engineered Klebsiella pneumoniae, which comprises the following steps:

[0053] Step 1, using the DNA of the genome sequence of Klebsiella ATCC BAA-830 (the preservation number of the NCBI database is NZ_CP010523.2) as a template, design primers, and the primers are designed as follows:

[0054] Kv-citT-F: GGCCGAATTCATGAAAGAGAAAAAGACAAC (as shown in SEQ ID NO: 1)

[0055] Kv-citT-R: GAGCGGATCCTCAGAACATCATGGACATCC (as shown in SEQ ID NO: 2),

[0056] The citT gene was completely cloned by PCR to obtain the citT gene.

[0057] Step 2: Use EcoRI and BamHI to digest the citT gene and the expression vector pDK6, connect the citT gene to the EcoRI and BamHI restriction sites of the expression vector pDK6, and construct the citT enhanced expression plasmid pDK6-citT.

[0058] Step 3: Prepare electroporation-competent Klebsiella pneumoniae ATCC 25955, transform the citT-enhanced ...

Embodiment 2

[0066] The present embodiment provides a construction method for enhanced expression of citT genetically engineered Klebsiella pneumoniae, which comprises the following steps:

[0067] Step 1, using the DNA of the genome sequence of Klebsiella pneumoniae ATCC 25955 (the preservation number of the NCBI database is AQQH00000000) as a template, design primers, and the primers are designed as follows:

[0068] Kp-citT-F: GAGCGGATCCATGAAAGAGAAAAAGACAAC (as shown in SEQ ID NO: 3)

[0069] Kp-citT-R: GGCCAAGCTTTCAGAACATCATGGACATCC (as shown in SEQ ID NO: 4),

[0070] The citT gene was completely cloned by PCR to obtain the citT gene. Through detection, the nucleotide sequence of the citT gene of Klebsiella pneumoniae ATCC25955 and the citT gene of Klebsiella mutans ATCC BAA-830 have 93% identity, and the amino acid sequence has 97% identity.

[0071] Step 2: Digest the citT gene and the expression vector pDK7 with BamHI and HindIII, connect the citT gene between the BamHI and HindI...

Embodiment 3

[0080] The present embodiment provides a construction method for enhanced expression of citT genetically engineered Klebsiella oxytoca, which comprises the following steps:

[0081] Step 1, using the DNA of the genome sequence of Klebsiella pneumoniae ATCC 25955 (the preservation number of the NCBI database is AQQH00000000) as a template, design primers, and the primers are designed as follows:

[0082] Kp-citT-F: GAGCGGATCCATGAAAGAGAAAAAGACAAC (as shown in SEQ ID NO: 3)

[0083] Kp-citT-R: GGCCAAGCTTTCAGAACATCATGGACATCC (as shown in SEQ ID NO: 4),

[0084] The citT gene was completely cloned by PCR to obtain the citT gene. Through detection, the nucleotide sequence of the citT gene of Klebsiella pneumoniae ATCC25955 and the citT gene of Klebsiella mutans ATCC BAA-830 have 93% identity, and the amino acid sequence has 97% identity.

[0085] Step 2: Digest the citT gene and the expression vector pDK7 with BamHI and HindIII, connect the citT gene between the BamHI and HindIII ...

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Abstract

The invention provides klebsiella for strengthening the expression of a citT gene and an application of the klebsiella in production of 1,3-propylene glycol. The klebsiella provided by the invention is specifically obtained by strengthening the expression of the citT gene in the klebsiella. By applying klebsiella provided by the invention to fermentation production of 1,3-propylene glycol, the yield of 1,3-propylene glycol can be increased.

Description

technical field [0001] The invention relates to a kind of Klebsiella bacteria expressing citT gene intensively and the application thereof for producing 1,3-propanediol. Background technique [0002] 1,3-propanediol is widely used and is a very important chemical product and platform compound. It can be used as adhesive, protective agent, lubricant, antifreeze and solvent in the fields of polymer materials, medicine, chemical industry, food and cosmetics have wide application. Its most important use is the production of polytrimethylene terephthalate (PTT). The good properties of PTT, including water washability, ductility and resilience, have made its market demand increasing. This drives the market demand for 1,3-propanediol (Liu H. et al., Biotechnol J. 2010, 5(11): 1137-1148). [0003] In view of the important role of 1,3-propanediol and the various disadvantages of chemical synthesis, the research on the production of 1,3-propanediol by microbial fermentation of glyc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/74C12P7/18C12R1/22
CPCC07K14/26C12N15/74C12P7/18
Inventor 韩冰
Owner ZHANGJIAGANG GLORY CHEM IND CO LTD
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