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Klebsiella bacteria with enhanced expression of tqsa gene and its application in the production of 1,3-propanediol

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

Active Publication Date: 2019-10-01
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 tqsA gene, but there is no research report on the function of Klebsiella tqsA gene

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The present embodiment provides a construction method for enhancing the expression of tqsA genetically engineered Klebsiella pneumoniae, which comprises the following steps:

[0054] 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:

[0055] Kv-tqsA-F: GGCCGAATTCATGGCTAAACCCATTATTAC (as shown in SEQ ID NO: 1)

[0056] Kv-tqsA-R: GAGCGGATCCTTATTCTTTGCTGACATCAC (as shown in SEQ ID NO: 2),

[0057] The tqsA gene was completely cloned by PCR to obtain the tqsA gene.

[0058] Step 2: Use EcoRI and BamHI to digest the tqsA gene and the expression vector pDK6, connect the tqsA gene to the EcoRI and BamHI restriction sites of the expression vector pDK6, and construct the tqsA enhanced expression plasmid pDK6-tqsA.

[0059] Step 3: Prepare electroporation-competent Klebsiella pneumoniae ATCC 25955, transform the tqsA enha...

Embodiment 2

[0067] The present embodiment provides a construction method for enhancing the expression of tqsA genetically engineered Klebsiella pneumoniae, comprising the following steps:

[0068] Step 1, using the DNA of the genome sequence of Klebsiella pneumoniae ATCC 25955 (the deposit number of the NCBI database is AQQH00000000) as a template, design primers, and the primers are designed as:

[0069] Kp-tqsA-F: GAGCGGATCCATGGCTAAACCCATTATTAC (as shown in SEQ ID NO: 3)

[0070] Kp-tqsA-R: GGCCAAGCTCTACTGTTTACTGACATCGC (as shown in SEQ ID NO:4),

[0071] The tqsA gene was completely cloned by PCR to obtain the tqsA gene. Through detection, the nucleotide sequence of the tqsA gene of Klebsiella pneumoniae ATCC25955 is 90% identical to the tqsA gene of Klebsiella mutans ATCC BAA-830, and the amino acid sequence is 98% identical.

[0072] Step 2, use BamHI and HindIII to digest the tqsA gene and the expression vector pDK7, and connect the tqsA gene to the BamHI and HindIII restriction s...

Embodiment 3

[0081] The present embodiment provides a construction method for enhancing the expression of tqsA genetically engineered Klebsiella oxytoca, which comprises the following steps:

[0082] Step 1, using the DNA of the genome sequence of Klebsiella pneumoniae ATCC 25955 (the deposit number of the NCBI database is AQQH00000000) as a template, design primers, and the primers are designed as:

[0083] Kp-tqsA-F: GAGCGGATCCATGGCTAAACCCATTATTAC (as shown in SEQ ID NO: 3)

[0084] Kp-tqsA-R: GGCCAAGCTCTACTGTTTACTGACATCGC (as shown in SEQ ID NO:4),

[0085] The tqsA gene was completely cloned by PCR to obtain the tqsA gene. Through detection, the nucleotide sequence of the tqsA gene of Klebsiella pneumoniae ATCC25955 is 90% identical to the tqsA gene of Klebsiella mutans ATCC BAA-830, and the amino acid sequence is 98% identical.

[0086] Step 2, use BamHI and HindIII to digest the tqsA gene and the expression vector pDK7, and connect the tqsA gene to the BamHI and HindIII restriction...

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PUM

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Abstract

The invention provides klebsiella for enhanced expression of a tqsA gene and application of the klebsiella in production of 1,3-propanediol. The klebsiella for enhanced expression of the tqsA gene is particularly obtained by enhanced expression of the tqsA gene in the klebsiella. The klebsiella for enhanced expression of the tqsA gene is used for producing the 1,3-propanediol in a fermented manner, and the yield of the 1,3-propanediol can be increased.

Description

technical field [0001] The invention relates to a Klebsiella spp. for enhancing the expression of tqsA gene and its application 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 in the production of polytrimethylene terephthalate (PTT). The good properties of PTT, including water washability, ductility and recovery, make its market demand expanding. As a result, the market demand for 1,3-propanediol is driven (Liu H. et al., Biotechnol J. 2010, 5(11): 1137-1148). [0003] In view of the important role of 1,3-propanediol and various disadvantages of chemical synthesis, the research on the production of 1,3-propanediol by microbial fermentation of glycerol has...

Claims

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Application Information

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