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Method for producing L-leucine and related DNA and microorganism

A technology of leucine and amino acids, applied in the field of DNA, can solve the relationship between L-leucine production ability of no L-leucine resistance and no L-leucine-resistant Escherichia spp. Bacteria and other issues

Inactive Publication Date: 2005-07-20
AJINOMOTO CO INC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, bacteria of the genus Escherichia that are resistant to L-leucine have not been found, nor has the relationship between L-leucine resistance and L-leucine production ability been found.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] Example 1: Obtaining DNA encoding mutant IPMS

[0103] (1) Obtaining L-leucine producing strain

[0104] The L-leucine-producing E. coli strain was obtained from the standard laboratory wild-type strain E. coli K-12 by the selection method described below. Escherichia coli K-12 strain was treated with a mutagen solution containing 0.2 mg / ml N-methyl-N-nitro-N-nitrosoguanidine at 37°C for 30 minutes. Then washed twice with sodium chloride solution (0.8%), plated on M9 agar medium containing 1 mg / ml L-leucine analogue 4-azoleucine. The colonies that appeared after incubation at 37°C for 5 days were picked to detect their ability to produce L-leucine. Strains No.9, No.68, No.58, No.55 and No.15 produced leucine.

[0105] (2) Obtaining the leuA gene from the L-leucine producing strain

[0106] The obtained L-leucine-producing mutant No.55 and the leuA gene of wild-type strain Escherichia coli K-12 were cloned by in vivo cloning using defective phage Mu d5005 (Groisman, ...

Embodiment 2

[0113] Example 2: Production of L-leucine by transformants

[0114] Escherichia coli C600 (leu -) (Appleyard R.K., Genetics, 39,440-452 (1954)) is used in producing L-leucine strain No.9, No.68, No.58, No.55 and No.15 and Escherichia coli K-12 Transduced with P1 phage grown in Get Leu + Transducers were tested for their ability to produce L-leucine. The data are shown in Table 3.

[0115] Donor strain

[0116] * L-leucine production was measured after 48 hours of incubation at 32°C. Data for leucine production represent the average of 10 transductants for each strain.

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PUM

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Abstract

A method for producing L-leucine, comprising the steps of: culturing a bacterium which is transformed with a DNA coding for an alpha-isopropylmalate synthase densensitized in feedback inhibition by L-leucine, in a culture medium to produce and accumulate L-leucine in the medium, and recovering L-leucine from the medium.

Description

Background of the invention [0001] The present invention relates to DNA encoding a mutant α-isopropylmalate synthase. The present invention also relates to an L-leucine-producing microorganism having a mutant α-isopropylmalate synthase, and a method for producing L-leucine using the microorganism. L-leucine is an essential amino acid that can be used as a food or feed nutritional additive, a reagent or material for medical treatment, pharmaceuticals or chemical industry, or a growth factor for the production of other amino acids such as lysine. [0002] In the past, L-leucine was mainly produced by a fermentation method using L-leucine-producing microorganisms belonging to the genus Brevibacterium, Corynebacterium, or Serratia, or mutants thereof (Amino acid fermentation, Japanese Society of Science Press , pp 397-422, 1986). [0003] The highest level of L-leucine accumulation was obtained with Brevibacterium flavum VKPM B-2736. The concentration of L-leucine accumulated i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/09C12N1/20C12N1/21C12N9/10C12N9/88C12N15/31C12N15/60C12N15/63C12P13/06C12R1/19
CPCC12N9/1025C12N9/88C12P13/06
Inventor M·M·古斯雅廷纳M·G·伦特斯Y·I·科兹洛夫L·V·伊万诺维斯卡雅E·B·沃罗希洛瓦
Owner AJINOMOTO CO INC
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