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Recombinant corynebacterium having 1,3-pdo productivity and reduced 3-hp productivity, and method for producing 1,3-pdo by using same

A 3-PDO, microbial technology, applied in the direction of microbial-based methods, biochemical equipment and methods, chemical instruments and methods, etc.

Pending Publication Date: 2021-07-23
KOREA ADVANCED INST OF SCI & TECH +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this is an effect caused by overexpression of exogenous enzymes, and among the aldehyde dehydrogenases that still naturally occur in C. glutamicum and participate in 3-HP biosynthesis, there is no connection with 3-HPA gene-related reports

Method used

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  • Recombinant corynebacterium having 1,3-pdo productivity and reduced 3-hp productivity, and method for producing 1,3-pdo by using same
  • Recombinant corynebacterium having 1,3-pdo productivity and reduced 3-hp productivity, and method for producing 1,3-pdo by using same
  • Recombinant corynebacterium having 1,3-pdo productivity and reduced 3-hp productivity, and method for producing 1,3-pdo by using same

Examples

Experimental program
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Effect test

Embodiment 1

[0048] In the following examples, only genes derived from specific strains are given as examples of genes to be introduced, but it will be apparent to those skilled in the art that any genes can be used without limitation as long as they are used in the genes to be introduced into them. It is enough to express in the host and exhibit the same activity. Example 1: Generation of the pCSglpFKD vector for the production of mutant C. glutamicum capable of growth using glycerol as a single carbon source

[0049] 1-1: Production of pCSglpFKD vector for constructing glycerol catabolism pathway

[0050] C. glutamicum is known to be unable to use glycerol as a single carbon source for cell growth. Therefore, to construct a glycerol catabolism pathway, a gene encoding an enzyme derived from Escherichia coli W3110 and responsible for a glycerol catabolism pathway was first expressed using the Corynebacterium glutamicum shuttle vector pCES208s-H36-S3.

[0051] Use the chromosomal DNA of ...

Embodiment 2

[0056] Example 2: Production of aldehyde dehydrogenase deletion vectors for inhibition of 3-HP biosynthesis

[0057] When 1,3-PDO is produced from glycerol, the precursor 3-HPA produced is converted to 3-HP by aldehyde dehydrogenase present in the cell. However, an enzyme catalyzing a reaction accepting a precursor as a substrate has not been reported in C. glutamicum. Therefore, in order to identify the aldehyde dehydrogenase that mediates the reaction and delete the gene encoding the enzyme from the genome of the strain to thereby inhibit 3-HP biosynthesis, the 13 aldehyde dehydrogenases present in Corynebacterium glutamicum were first selected. Hydrogenase (Table 2).

[0058] Then, in order to confirm the inhibitory effect on 3-HP biosynthesis by deleting the genes (SEQ ID NO: 56 to 68) encoding these 13 aldehyde dehydrogenases, pTacCC1- Strains of the HrT vector (Cho et al., Metabolic Engineering, 42:157-167, 2017). Then, i) pCG9ts series each containing sgRNA sequences...

Embodiment 3

[0078] Example 3: Production and confirmation of Corynebacterium glutamicum with suppressed 3-HP production capacity and improved 1,3-PDO production capacity

[0079] 3-1: Production of Corynebacterium glutamicum having suppressed 3-HP production ability

[0080] The pCG9ts-ALD vector and ssODN produced in Examples 2-1 and 2-2 were each transformed into wild-type Corynebacterium glutamicum (ATCC 13032) in order to delete from the genome any coding sequence expected to be involved in 3-HP biosynthesis. Genes for 13 aldehyde dehydrogenases. Then, for the transformed mutant Corynebacterium glutamicum strain, the pTacCC1-HrT vector was removed by solidification on a 37°C BHI plate (Korean Patent Application No. 2017-0042124; Cho et al., Metabolic Engineering, 42:157-167, 201, SEQ ID NO:57) and pCG9ts-ALD vector. The strains produced by this process are shown in Table 6. However, the WAH3 and WAH9 strains were not produced, and the corresponding two genes were considered essenti...

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Abstract

The present invention relates to recombinant Corynebacterium having 1,3-PDO productivity and reduced 3-HP productivity, and a method for producing 1,3-PDO by using same. When a Corynebacterium glutamicum variant according to the present invention is used, the productivity of 3-HP, which is a by-product, is inhibited by using low-cost glycerol as a carbon source, and thus 1,3-PDO can be produced with high efficiency.

Description

technical field [0001] The present invention relates to a recombinant Corynebacterium having 1,3-PDO (1,3-propanediol)-producing ability and reduced (suppressed) 3-HP-producing ability and a method for producing 1,3-PDO using the same, And more particularly to a mutant microorganism having a missing or weakened 3-HP production ability and producing 1,3-PDO from glycerol, wherein the mutant microorganism is produced by translating a protein encoding a glycerol-facilitating protein Genes, genes encoding glycerol kinase, genes encoding glycerol dehydrogenase, genes encoding glycerol dehydratase, genes encoding glycerol reactivation enzyme, and genes encoding 1,3-PDO oxidoreductase were introduced into Corynebacterium glutamicum (Corynebacterium glutamicum), and the gene encoding aldehyde dehydrogenase was deleted or attenuated from said Corynebacterium glutamicum. Background technique [0002] 1,3-Propanediol (1,3-PDO) is a chemical used as a monomer for the synthesis of polym...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/77C12N15/67C12N9/04C12N9/02C12N9/12C12P7/18C07K14/34
CPCC12N15/77C12P7/18C12Y207/0103C12Y402/0103C12Y101/01202C12Y101/01006C12N15/52C12Y102/01003C12N9/1205C12N9/0006C12N9/0008C07K14/245C12R2001/15C12Y101/05003C12Y101/01008C12N9/88C12Y102/01008C12Y101/05004C12N15/67C07K14/34
Inventor 李相烨赵在星金渧雄C·P·S·普拉博沃韩泰熙高裕诚
Owner KOREA ADVANCED INST OF SCI & TECH
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