Method for producing heavy chain aminocarboxylic acid

A transaminase gene and gene technology, applied in the field of production of medium-chain aminocarboxylic acids, can solve the problems of high yield production and other problems

Inactive Publication Date: 2018-08-31
KOREA RES INST OF BIOSCI & BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since medium-chain aminocarboxylic acids are produced by further introducing a process for transferring an amine group to medium-chain aldehydecarboxylic acids, one disadvantage of such medium-chain aminocarboxylic acids is that when microorganisms are used to produce the medium-chain aminocarboxylic acids cannot be produced at high yields when

Method used

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  • Method for producing heavy chain aminocarboxylic acid
  • Method for producing heavy chain aminocarboxylic acid
  • Method for producing heavy chain aminocarboxylic acid

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Experimental program
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Embodiment approach

[0029] In order to achieve the purpose of the present invention, the present invention provides a recombinant microorganism, wherein the fatty aldehyde dehydrogenase gene in the ω-oxidative metabolic pathway and the genes related to the β-oxidative metabolic pathway are deleted, and the ω-transaminase gene is also introduced therein.

[0030] In the present invention, the term "ω-oxidation" refers to a metabolic process in which the terminal methyl group of a fatty acid is oxidized to form a dicarboxylic acid, and the term "β-oxidation" refers to a metabolic process in which the carbon at the β position of the carboxyl group Atoms are oxidized to release acetyl-CoA, whereby fatty acids are gradually decomposed into fatty acids with a reduced number of carbon atoms by 2. The concepts of ω-oxidation and β-oxidation and the enzymes involved in this metabolic process are well known to those of ordinary skill in the field of biochemistry. For example, when fatty acids are used as s...

example 1

[0050] Example 1: Construction of a knockout cassette

[0051] A vector was constructed containing the ura3 gene used as a selection marker for gene knockout for changing strains and a pop-up region for deletion of the ura3 gene after insertion of the knockout cassette ( image 3 ). A gene derived from Yarrowia was used as the ura3 gene, and the pop-up region for changing the strain had a total of four sequences, and two genes were referred to. Here, a glutamic acid production gene derived from Bacillus was used as one of the genes, and a His operon-related gene derived from Salmonella or the cloning vector pHUKH was used as the other gene. Table 1 below lists the primers and sequences used to construct the pop-out vector.

[0052] 【Table 1】

[0053]

[0054] like Figure 4 Knockout cassettes were constructed as indicated. First, PCR was performed on the homology region (HR) to be knocked out from the Yarrowia genomic DNA, and on the two 5' and 3' end fragments in th...

Embodiment 2

[0062] Embodiment 2: Construction of transduction vector

[0063] To insert an omega-transaminase into a Yarrowia strain, constructs such as Figure 5 vector shown. Table 4 lists the primers used for this purpose.

[0064] 【Table 4】

[0065]

[0066] with Figure 4 The transaminase cassette was constructed in the same manner except that when two PCR product fragments were obtained from the vector, the gene from the promoter to ura3 was amplified to construct the cassette. Primers used to construct the cassettes are listed in Table 5 below.

[0067] 【table 5】

[0068]

[0069] The gene sequences used to alter the recombinant microbial strains according to the present invention are listed in the sequence listing and summarized in Table 6.

[0070] 【Table 6】

[0071] Gene

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Abstract

The present invention relates to a method for producing a heavy chain aminocarboxylic acid and, more particularly, to a recombinant microorganism in which a fatty aldehyde dehydrogenase gene in [omega]-oxidative metabolic pathway and a [beta]-oxidative metabolic pathway-related gene are deleted and a [omega]-transaminase gene is introduced, and a method for producing a heavy chain aminocarboxylicacid by culturing the recombinant microorganism. The recombinant microorganism of the present invention can prevent additional oxidation of fatty aldehyde and [beta]-oxidative metabolism and also produce a heavy chain aminocarboxylic acid, such as 12-aminodecane, as a raw material of nylon 12 from a substrate such as fatty acid with a high yield by introducing an amine group into a terminal thereof.

Description

technical field [0001] The present invention relates to a method for producing medium-chain aminocarboxylic acids, and more particularly to a method for producing medium-chain aminocarboxylic acids from fatty acids by culturing a recombinant microorganism in which fatty aldehyde dehydrogenase ( or fatty alcohol dehydrogenase) gene and β-oxidation metabolic pathway-related genes were deleted, and ω-transaminase gene was also introduced into it. Background technique [0002] Bio-platform compounds are produced based on biomass-derived raw materials through biological or chemical transformation, and thus have been used to synthesize polymer monomers, new materials, etc. [0003] Among bioplatform compounds, medium-chain aminocarboxylic acids are materials used as polyamide monomers. Polyamides are classified into aliphatic polyamides, aromatic polyamides and aliphatic cyclic polyamides. Representative examples of aliphatic polyamides include nylon 12, nylon 6, and nylon 66, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C12N15/81C12P7/64C12P13/00C12N9/02C12N9/10
CPCC12P7/6409C12P13/04C12N9/0008C12Y102/01003C12Y206/01018C12N9/1096C12Y206/01C12N9/001C12N15/70C12N15/81C12P13/001C12Y103/03006C12N15/815C12P13/005
Inventor 安廷梧李弘遠朴奎妍张旼祯全宇荣
Owner KOREA RES INST OF BIOSCI & BIOTECH
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