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Genetically modified microorganism for producing 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and/or hexanedioic acid, and production method for the chemicals

A technology of adipic acid, hydroxyadipic acid, applied to genetically modified microorganisms for the production of 3-hydroxyadipic acid, alpha-hydrogenated adipic acid and/or adipic acid and the chemical In the field of manufacturing, it can solve problems such as no pyruvate kinase

Pending Publication Date: 2021-12-14
TORAY IND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this document, although it is specifically disclosed that the activity of acyltransferase that catalyzes the reaction of 3-oxoadipyl-CoA from acetyl-CoA and succinyl-CoA can be enhanced, 3-hydroxyadipate can be increased. and α-hydrogenated hexadienate production efficiencies, but nothing about pyruvate kinase

Method used

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  • Genetically modified microorganism for producing 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and/or hexanedioic acid, and production method for the chemicals
  • Genetically modified microorganism for producing 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and/or hexanedioic acid, and production method for the chemicals
  • Genetically modified microorganism for producing 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and/or hexanedioic acid, and production method for the chemicals

Examples

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preparation example Construction

[0128] Preparation of 3-oxoadipyl-CoA solution: According to conventional methods, PCR was carried out with the genomic DNA of Pseudomonas putida KT2440 strain as a template, and the nucleic acids encoding CoA transferase (pcaI and pcaJ, NCBI-Gene ID: 1046613 and 1046612) were amplified in full length. In addition, the base sequences of the primers used for this PCR are SEQ ID NO: 194 and 195, for example. This amplified fragment was inserted into the KpnI site of pRSF-1b (manufactured by Novagen), which is an expression vector for Escherichia coli, so as to form the same frame as the histidine tag sequence. The plasmid was introduced into Escherichia coli BL 21 (DE3), and the expression of the enzyme was induced by using isopropyl-β-thiogalactopyranoside (IPTG) according to the conventional method, followed by the use of the histidine tag from the culture medium. Refined to obtain a CoA transferase solution. Using this solution, an enzyme reaction solution for producing 3-o...

Embodiment

[0240] Hereinafter, an Example is given and this invention is demonstrated concretely.

reference example 1

[0242] For the expression of enzymes that catalyze the reaction to form 3-oxoadipyl-CoA and coenzyme A (reaction A), the reaction to 3-hydroxyadipate from 3-hydroxyadipyl-CoA (reaction E ) and enzymes that catalyze the reaction (reaction F) to generate α-hydroadipiendioic acid from 2,3-dehydroadipyl-CoA, and Sequence numbers 1, 2, 3, 4, 5, 6, Production of the plasmid of the polypeptide described in 7

[0243] The self-replicating vector pBBR1MCS-2 in Escherichia coli (ME Kovach, (1995), Gene 166:175-176) was excised with XhoI to obtain pBBR1MCS-2 / XhoI. In order to assemble the constitutive expression promoter in this vector, the genomic DNA of Escherichia coli K-12MG1655 was used as a template to design the upstream region 200b (SEQ ID NO: 186) of gapA (NCBI Gene ID: NC000913.3). The primers (sequence numbers 187 and 188) for PCR amplification were subjected to PCR reaction according to conventional methods. The resulting fragment was ligated with pBBR1MCS-2 / XhoI using In-F...

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Abstract

The invention discloses a genetically modified microorganism capable of producing 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and / or hexanedioic acid at a high yield, and a production method for 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and / or hexanedioic acid using the genetically modified microorganism. The genetically modified microorganism has the ability to produce 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and / or hexanedioic acid, and disrupts the function of pyruvate kinase and enhances the activity of phosphoenolpyruvate carboxykinase and the activity of enzymes that catalyze a reaction for generating 3-hydroxyadipyl-CoA by reducing 3-oxoadipyl-CoA.

Description

technical field [0001] The present invention relates to a genetically modified microorganism that produces 3-hydroxyadipic acid, α-hydrogenated adienedioic acid, and / or adipic acid at high yields, and to producing 3-hydroxyadipic acid, α-hydrogenated adipic acid, and / or adipic acid using the genetically modified microorganism. - A process for the hydrogenation of adienedioic and / or adipic acid. Background technique [0002] 3-Hydroxyhexanedioic acid (IUPAC name: 3-hydroxyhexanedioic acid) and α-hydrogenated hexanedioic acid (IUPAC name: (E)-hex-2-enedioic acid) are dicarboxylic acids having 6 carbon atoms. These can be used as raw materials for polymerizing with polyols to obtain polyesters, or polymerizing with polyamines to obtain polyamides. In addition, compounds obtained by adding ammonia to their terminals to be lactamized can be used as raw materials for polyamides. [0003] The following documents are known concerning the production of 3-hydroxyadipic acid or α-hyd...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/42C12N5/10C12N1/15C12N1/19C12N1/21C12N9/02
CPCC12P7/42C12P7/44C12N9/0006C12Y101/01035C12Y401/01049C12Y207/0104C12N9/88C12N1/20C12N9/1217C12R2001/425C12N5/10
Inventor 矶部匡平河村健司山田胜成
Owner TORAY IND INC
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