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Microorganism capable of producing quinolinic acid, and method for producing quinolinic acid utilizing the microorganism

A technology of microorganisms and quinolinic acid, applied in the field of microorganisms capable of producing quinolinic acid and using the microorganisms to produce quinolinic acid, which can solve problems such as the impact of oil extraction costs and environmental problems

Inactive Publication Date: 2017-10-20
CJ CHEILJEDANG CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since chemical synthesis methods use non-renewable materials derived from petroleum as raw materials, there is a problem that they are greatly affected by environmental problems, oil prices, or oil extraction costs

Method used

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  • Microorganism capable of producing quinolinic acid, and method for producing quinolinic acid utilizing the microorganism

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1. Preparation produces the bacterial strain of quinolinic acid

[0073] 1-1. Preparation of quinolinic acid phosphoribosyltransferase-depleted strain

[0074] The following experiments were performed to delete the nadC gene encoding the quinolinic acid phosphoribosyltransferase involved in the quinolinic acid degradation pathway.

[0075] The nadC gene of the quinolinic acid degradation pathway was obtained by performing PCR using the chromosomal DNA of Escherichia coli K12W3110 as a template. Nucleotide sequence information of nadC gene (NCBI accession number "GI:89106990", SEQ ID NO:4) was obtained from GenBank of National Institutes of Health (NIH), and its amino acid sequence was identical to SEQ ID NO:3. Based on SEQ ID NO: 4, the primers of SEQ ID NO: 5 and the primers of SEQ ID NO: 6 for amplifying the downstream region of nadC gene were synthesized, the primers of SEQ ID NO: 7 and the primers of SEQ ID NO: 7 for amplifying the upstream and downstrea...

Embodiment 2

[0120] Example 2. Evaluation of the productivity of quinolinic acid producing strains

[0121] To evaluate the ability of the nadA(KP) gene-enhanced strain to produce quinolinic acid, the pNadA-nadA vector and the pNadA-nadA(KP) vector were introduced into the W3110-ΔnadC strain, and the W3110ΔnadC / pNadA-nadA strain and the W3110ΔnadC / pNadA-nadA (KP) strains were named CV01-0812 and CV01-0813. Furthermore, to enhance nadB, the pPro-nadB vector was introduced into CV01-0812 and CV01-0813, and they were named CV01-0814 and CV01-0815, respectively.

[0122] Vector introduction method is using CaCl 2 Method transformation, and in the incubator of 37 ℃, CV01-0812 and CV01-0813 are in LB-Sp (10g / L yeast extract, the NaCl of 5g / L, the tryptone of 10g / L, the agar of 1.5% , 50 μg / L of spectinomycin) plate medium and culture overnight, CV01-0814 and CV01-0815 in LB-sp, Km (yeast extract of 10g / L, NaCl of 5g / L, 10g / L of tryptone, 1.5% agar, 50 μg / L of kanamycin, 50 μg / L of spectinomy...

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Abstract

The present invention relates to a microorganism capable of producing quinolinic acid, and to a method for producing same utilizing the microorganism.

Description

technical field [0001] The present disclosure relates to microorganisms capable of producing quinolinic acid and methods of producing quinolinic acid using the microorganisms. Background technique [0002] Quinolinic acid is known as 2,3-pyridinedicarboxylic acid, and is used as a precursor of chemical compounds in various fields such as medicine, agricultural chemicals, and dyeing materials. [0003] Quinolinic acid can be prepared by chemical synthesis method or biosynthesis method. Since the chemical synthesis method uses non-renewable materials derived from petroleum as a raw material, it has a problem of being greatly affected by environmental problems, oil prices, or oil extraction costs. [0004] As a representative example of the biosynthetic method, there is a method for producing quinolinic acid in which genes encoding L-aspartate oxidase (NadB) and quinolinic acid synthase (NadA) are cloned into each having a different copy number , and after enhancing the expre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/70C12P7/46C12R1/19
CPCC12N9/10C12N15/70C12P7/46C12N9/0022C12N9/1085C12Y104/03016C12Y205/01072C12P17/12
Inventor 李在旼金朱恩李在熙张真淑金素影申容旭
Owner CJ CHEILJEDANG CORP
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