Microorganism producing l-methionine precursor and the method of producing l-methionine precursor using the microorganism

A technology of methionine and microorganisms, which is applied in the field of microorganisms for preparing L-methionine precursor O-succinylhomoserine, and can solve the problems of small methionine yield and the like

Active Publication Date: 2009-10-14
CJ CHEILJEDANG CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, the production of methionine is relatively small

Method used

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  • Microorganism producing l-methionine precursor and the method of producing l-methionine precursor using the microorganism
  • Microorganism producing l-methionine precursor and the method of producing l-methionine precursor using the microorganism
  • Microorganism producing l-methionine precursor and the method of producing l-methionine precursor using the microorganism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1: Construction of methionine precursor production strain

[0068] Deletion of the metB gene

[0069] To delete the metB gene encoding cystathionine synthase in E. coli, a FRT-one-step PCR deletion was performed (PNAS (2000) vol 97: p6640-6645). Primers of SEQ ID NO: 1 and NO: 2 were used for PCR using pKD3 vector (PNAS (2000) vol 97: p6640-6645) as a template to obtain the construction of the deletion gene cassette. PCR was performed as follows: 30 cycles of denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, and extension at 72°C for 1 minute.

[0070]The PCR product was subjected to 1.0% agarose gel electrophoresis, followed by purification of the DNA obtained from the 1.2 kbp band. The recovered DNA fragment was electroporated into Escherichia coli (K12) W3110 transformed with pKD46 vector (PNAS (2000) vol 97: p6640-6645). Before electroporation, W3110 transformed with pKD46 was grown to OD at 30°C in LB medium containing 100 μg / ...

Embodiment 2

[0107] Example 2: Fermentation for the production of L-methionine precursor

[0108] Glass bottle culture experiment

[0109] In order to examine the productivity of the methionine precursor of each strain constructed in Example 1, Erlenmeyer flask culture was used. Each strain was cultured overnight at 31°C in LB plate medium. Single clones were inoculated into 3 ml of LB medium, followed by culturing at 31°C for 5 hours. The culture solution was diluted 200 times into a 250ml Erlenmeyer flask containing 25ml of methionine precursor production medium, and then cultured at 31°C and 200rpm for 64 hours. HPLC was performed to compare the methionine precursor production capacity (Table 2 and Table 3). As a result, the methionine-producing ability was significantly increased in the methionine precursor-producing strain prepared from the L-threonine-producing strain having no methionine requirement.

[0110] Table 1

[0111] Glass bottle media components for methionine pr...

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Abstract

The present invention relates to a microorganism producing L-methionine precursor, O-succinylhomoserine, 1) the homoserine O-succinyltransferase activity (EC2.3.1.46) is introduced and enhanced, wherein the homoserine O-succinyltransferase activity is feed back resistant to methionine; and 2) the aspartokinase or homoserine dehydrogenase activity (EC2.7.2.4 or 1.1.1.3) is enhanced, and a method of producing L-methionine precursor using the microorganism.

Description

technical field [0001] The present invention relates to a microorganism for producing L-methionine precursor O-succinyl homoserine, and a method for producing L-methionine precursor using the microorganism. Background of the invention [0002] Methionine, one of the essential amino acids for the human body, is widely used as a feed and food additive, and also as a synthetic raw material for medical solutions and medical appliances. Methionine is a precursor to compounds such as choline (lecithin) and creatine, and is also used as a raw material for the synthesis of cysteine ​​and taurine. Methionine also provides sulfur. S-adenomethionine is derived from L-methionine and plays a role in donating methyl groups in humans and is also involved in the synthesis of various neurotransmitters in the brain. Methionine and / or S-adeno-L-methionine (SAM) inhibit fat accumulation in the liver and arteries and can reduce depression, inflammation, liver disease and muscle pain, among oth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12P13/06C12R1/185C12R1/19
CPCC12P13/06C12N9/1029
Inventor 申容旭金素影张真淑赵英郁李汉珍许仁庚罗光镐徐昌一金喆河严惠媛
Owner CJ CHEILJEDANG CORP
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