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Recombinant expression of aldehyde reductase and application thereof in bioconversion of glycerol into 1,3-propylene glycol

A biotransformation and reductase technology, applied in the field of bioengineering, can solve problems such as ineffective utilization, and achieve the effect of increasing yield

Inactive Publication Date: 2011-09-14
DALIAN UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Considering that there is a large amount of NADPH in the bacteria, and some microorganisms, such as Klebsiella (Klebsiella), Citrobacter (Citrobacter), Clostridium (Clostridium), etc., 1,3 -Propanediol oxidoreductase cannot efficiently utilize NADPH to synthesize 1,3-propanediol

Method used

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  • Recombinant expression of aldehyde reductase and application thereof in bioconversion of glycerol into 1,3-propylene glycol
  • Recombinant expression of aldehyde reductase and application thereof in bioconversion of glycerol into 1,3-propylene glycol
  • Recombinant expression of aldehyde reductase and application thereof in bioconversion of glycerol into 1,3-propylene glycol

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Experimental program
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Embodiment

[0024] (1) Cloning of aldehyde reductase gene yqhD:

[0025] Primers were designed according to the aldehyde reductase gene sequence (yqhD, ID: EU012494) of Klebsiella pneumoniae AS1.1736 published in GenBank as follows:

[0026] P1: 5'-ATGAATAATTTCGACCTGCATACCC-3' (SEQ ID NO: 3)

[0027] P2: 5'-TTAGCGTGCAGCCTCGTAAATAC-3' (SEQ ID NO: 4)

[0028] Complete PCR reaction (referring to Sambrook, J ., Russel, D.W., Molecular Cloning: A Laboratory Maunal 3rd ed., Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press, 2001). Add the following components to the PCR reaction tube: 1 μl of genomic DNA, 4 μl of dNTPs, 1 μl of primers P1 and P2 (synthesized by Treasure Bioengineering (Dalian) Co., Ltd.), 5 μl of 10×Ex Taq buffer, 1U of Ex Taq DNA polymerase, and make up to 50 μl of water; Reaction conditions: Denaturation at 95°C for 60s, 30 cycles at 95°C for 30s, 58°C for 1min, and 72°C for 2min. The obtained PCR product was confirmed by electrophoresis analysis, purified...

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Abstract

The invention discloses recombinant expression of aldehyde reductase and an application thereof in bioconversion of glycerol into 1, 3-propylene glycol, belonging to the technical field of biological engineering. The invention is characterized in that a recombinant expression vector containing aldehyde reductase gene sequence is used to express the enzyme to a host cell for preparing 1, 3-propylene glycol. The beneficial effects and benefits of the invention are that recombinant Klebsiella (Pdk-yqhD) takes glycol as substrate to ferment for 30h, and the concentration of 1, 3-propylene glycol in the fermentation liquor is improved by 17.1% in comparison with a comparison strain; compared with the original strain, by using the recombinant restrain in the invention for fermentation, the yield of 1, 3-propylene glycol is improved by 10-50%, which lays a foundation on constructing genetically engineered microorganism of the high yield of the 1, 3-propylene glycol taking glycol as substrate.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a recombinant expression vector containing an aldehyde reductase gene and the application of a host cell transformed with the recombinant expression vector to ferment glycerol to high-yield 1,3-propanediol. Background technique [0002] 1,3-propanediol (1,3-propanediol, referred to as 1,3-PD) is a colorless, odorless viscous liquid, is an important chemical raw material, can be used as a solvent, antifreeze or protective agent, fine Chemical raw materials and new polyesters - polytrimethylene terephthalate (PTT) and polyurethane monomers. PTT is a new type of polyester material, which can be used in carpets, engineering plastics, films and clothing materials in the textile industry, etc., and PTT has excellent resilience, dyeability, stain resistance, good anti-ultraviolet discoloration performance and is not easy to rise It has the characteristics of static electricity and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/53C12N15/74C12N1/21C12N9/02C12P7/18C12R1/01C12R1/145C12R1/22
Inventor 修志龙张乐马成伟戴建英
Owner DALIAN UNIV OF TECH
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