Recombinant microorganism of genus escherichia with l-threonine productivity, and method for producing l-threonine using same

A technology of microorganisms and threonine, applied in the direction of recombinant DNA technology, bacteria, peptides, etc., can solve the problems of no production relationship, high homology of galP gene, etc., and achieve improved productivity, high yield, and increased growth speed Effect

Active Publication Date: 2016-10-12
CJ CHEILJEDANG CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] It has been reported that inositol permeases encoded by iolT1 and iolT2 genes also function as glucose permeases in Corynebacterium glutamicum (Ikeda et al., Appl Microbiol Biotechnol (2011) 90:1443-1451); The gene has a high homology with the galP gene of Escherichia coli, however, there is no report showing the relationship between the inositol permease and threonine production

Method used

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  • Recombinant microorganism of genus escherichia with l-threonine productivity, and method for producing l-threonine using same
  • Recombinant microorganism of genus escherichia with l-threonine productivity, and method for producing l-threonine using same
  • Recombinant microorganism of genus escherichia with l-threonine productivity, and method for producing l-threonine using same

Examples

Experimental program
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preparation example Construction

[0039] The present invention also provides a preparation method of L-threonine, which is characterized by comprising the following steps: cultivating the transformed recombinant microorganism belonging to the genus Escherichia; and isolating L-threonine from the microorganism culture solution.

[0040] Cultivation of the recombinant microorganism belonging to the genus Escherichia of the present invention can be carried out by a conventional method. Specifically, raw sugar can be contained in whole or in part or glucose as a carbon source medium to inoculate the microorganisms and culture. The culturing process can be performed under appropriate media and culture conditions known in the art. Such culture procedures can be easily adjusted and used by those skilled in the art according to the selected strain. Examples of the culture method include batch culture, continuous culture and fed-batch culture, but are not limited thereto. The medium used in the cultivation should s...

Embodiment 1

[0066] Example 1: Production of recombinant vectors comprising iolT1 and iolT2 genes derived from coryneform bacteria

[0067] (1) Homology comparison with Escherichia coli glucose permease (galP)

[0068] It has been reported that the iolT1 and iolT2 genes encoding inositol permease in Corynebacterium glutamicum have similar homology to the galP gene encoding glucose permease in Escherichia coli. From the genome of wild-type Corynebacterium glutamicum ATCC 13032, a gene homologous to the galP gene derived from Escherichia coli was found and compared, and the results are shown in figure 1 middle.

[0069] The amino acid sequence encoded by galP derived from Escherichia coli showed 34% homology with the amino acid sequence encoded by iolT1 derived from Corynebacterium glutamicum, and showed 31% homology with the amino acid sequence encoded by iolT2.

[0070] (2) Production of iolT1 gene fragment

[0071] In order to obtain a 1.5kb fragment comprising the open reading frame...

experiment example 2

[0085] Experimental example 2: Preparation of transformed recombinant strains and comparison of L-threonine productivity

[0086] (1) Preparation of recombinant strains utilizing wild-type Escherichia coli

[0087] The recombinant vectors pCC1BAC-iolT1, pCC1BAC-iolT2, and pCC1BAC-iolT1-iolT2 prepared in Example 1 were introduced into the overexpression vector pBRThrABCR3 (LeeKH et al., Molecular Systems In the wild-type Escherichia coli MG1655 of Biology (2007) 3:149), the Escherichia coli was spread on a solid medium containing 100 μg / ml ampicillin and 15 μg / ml chloramphenicol, and a single colony was selected.

[0088] The selected strains were named MG1655 / pBRThrABCR3 / pCC1BAC-iolT1, MG1655 / pBRThrABCR3 / pCC1BAC-iolT2 and MG1655 / pBRThrABCR3 / pCC1BAC-iolT1-iolT2, respectively.

[0089] The L-threonine productivity of these strains was confirmed by the same method as (3) of Comparative Example 1 using the threonine titer medium prepared according to the composition of Table 1 ...

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Abstract

The present invention relates to an Escherichia coli mutant strain having improved L-threonine productivity by the introduction of permease derived coryneform bacteria, and a method for producing L-threonine using the same.

Description

technical field [0001] The present invention relates to a recombinant microorganism belonging to the genus Escherichia having improved productivity of L-threonine by deforming to express a permease derived from a coryneform bacterium and a method for producing L-threonine using the microorganism. Background technique [0002] L-threonine is an essential amino acid, which is widely used as feed and food additives, and as a synthetic raw material for hydration solutions of medicines and medicines. [0003] L-threonine is mainly produced by a fermentation method using bacteria of the genus Escherichia, Serratia, Provi Denstella or Corynebacterium. Genes related to the biosynthesis of threonine and various methods for increasing the expression of these genes have been developed, but a method capable of producing L-threonine at a high yield in a cheaper manner is still required. [0004] It is known that the GalP protein encoded by the galP gene in Escherichia coli is a galacto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/52C12P13/08
CPCC07K14/34C12P13/08C12N15/52
Inventor 金亨俊权秀渊高恩圣李知宣李根喆黄荣彬洪亨杓
Owner CJ CHEILJEDANG CORP
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