Recombined bacterial strain for modifying specificity of 1,3-propanediol redoxase coenzyme and application thereof

A technology of reductase and propylene glycol, which is applied in the field of bioengineering, can solve the problem that propylene glycol oxidoreductase cannot obtain the expected effect, and achieve the effect of increasing production

Inactive Publication Date: 2013-04-10
DALIAN UNIV OF TECH
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Problems solved by technology

But even if the 1,3-propanediol oxidoreductase is highly expressed in the bacteria, the expected effect cannot be obtained (Zheng, P et al., Process Biochem.2006, 41(10): 2160-2169)

Method used

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  • Recombined bacterial strain for modifying specificity of 1,3-propanediol redoxase coenzyme and application thereof
  • Recombined bacterial strain for modifying specificity of 1,3-propanediol redoxase coenzyme and application thereof
  • Recombined bacterial strain for modifying specificity of 1,3-propanediol redoxase coenzyme and application thereof

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Embodiment Construction

[0028] Specific embodiments of the present invention will be described in detail below in conjunction with technical solutions.

[0029] In the following specific embodiments, the rational design method of molecular simulation is first used to find the key amino acid residues that affect the binding specificity of the 1,3-propanediol oxidoreductase coenzyme, and the method of primer overlap extension site-directed mutagenesis is used to realize the mutation transformation, and then the construction containing The expression vector of the mutant enzyme and the recombinant bacterial strain containing the expression vector are finally used to ferment glycerol to produce 1,3-propanediol with the constructed recombinant bacterial strain.

[0030] 1. Rational design of 1,3-propanediol oxidoreductase coenzyme binding specificity:

[0031] Rational design of coenzyme binding specificity is accomplished through a series of molecular simulation methods. First, homology modeling was use...

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Abstract

The invention relates to a recombined bacterial strain for modifying the specificity of a 1,3-propanediol redoxase coenzyme and an application of the recombined bacterial strain to 1,3-propanediol preparation, belonging to the technical field of biological engineering. The recombined bacterial strain is characterized in that the specificity of the 1,3-propanediol redoxase coenzyme is modified by rational design to obtain a mutant enzyme; and a recombinant expression vector in a gene sequence containing the mutant enzyme is constructed and converted into a host cell to obtain a recombined bacterial strain. Compared with an original bacterial strain, the invention has the advantage that the 1,3-propanediol yield is improved by 10-50 percent by utilizing the recombined bacterial strain for fermentation.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to the rational design and transformation of the coenzyme specificity of 1,3-propanediol oxidoreductase, and also relates to a recombinant expression vector containing the gene sequence of the mutant enzyme, and the use of host cells containing the vector in Application in fermenting glycerol to produce 1,3-propanediol. Background technique [0002] 1,3-propanediol (1,3-propanediol, referred to as 1,3-PD) is a colorless, odorless viscous liquid. As an important chemical raw material, 1,3-propanediol can be used as solvent, antifreeze or protective agent, fine chemical raw material and monomer of new polyester-polytrimethylene terephthalate (PTT) and polyurethane. 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, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/53C12N15/74C12N9/04C12N1/21C12P7/18C12R1/01C12R1/22
CPCY02P20/52
Inventor 修志龙马成伟张乐戴建英
Owner DALIAN UNIV OF TECH
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