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L-leucine high-yield bacterium and fermentation method using the same for L-leucine production

A technology of Leucine and Corynebacterium glutamicum, applied in the field of bioengineering

Active Publication Date: 2005-11-30
WUXI JINGHAI AMINO ACID
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The acid production level of the L-leucine production bacteria reported above is still far from industrial production. At present, there is no L-leucine product produced by fermentation in my country's amino acid manufacturers, and the L-leucine product sold in the domestic market Amino acids are all produced by extraction

Method used

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  • L-leucine high-yield bacterium and fermentation method using the same for L-leucine production

Examples

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Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Preparation of mutant strains resistant to leucine hydroxamate.

[0020] The starting strain Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC 13032 was subjected to routine mutagenesis treatment with diethyl sulfate (1%, 30°C, 30 minutes), and then coated with 5-10g / L leucine oxygen On the basic medium of hydroxamate, culture at 30°C for 2 to 4 days, pick the colony grown on this medium as a mutant strain resistant to leucine hydroxamate, and then carry out L-leucine Acid fermentation test, among these mutants, the mutant with the highest L-leucine production is called Corynebacterium glutamicum L1-68, and its L-leucine production is 8.9 g / L.

Embodiment 2

[0021] Example 2 Preparation of mutants resistant to 2-thiazolealanine.

[0022] Corynebacterium glutamicum (Corynebacterium glutamicum) L1-68 obtained in Example 1 was subjected to routine mutagenesis treatment with diethyl sulfate (1%, 30° C., 25 minutes), and then coated in a solution containing 5-10 g / L On the basic medium of 2-thiazolealanine, culture at 30°C for 2 to 4 days, pick the colonies grown on this medium as mutants resistant to 2-thiazolealanine, and then carry out L- Leucine fermentation test, among these mutants, the mutant with the highest L-leucine production is called Corynebacterium glutamicum L2-132, and its L-leucine production is 13 g / L.

Embodiment 3

[0023] Example 3 Preparation of mutant strains resistant to α-aminobutyric acid.

[0024] The Corynebacterium glutamicum (Corynebacterium glutamicum) L2-132 obtained in Example 2 was subjected to routine mutagenesis treatment with nitrosoguanidine (0.3%, 30° C., 45 minutes), and then coated in a solution containing 20-40 g / L On the basic medium of α-aminobutyric acid, culture at 30°C for 2 to 4 days, pick the colonies grown on this medium as mutants resistant to α-aminobutyric acid, and then carry out L-leucine Acid fermentation test, among these mutant strains, the mutant strain with the highest L-leucine production is called Corynebacterium glutamicum L3-206, and its L-leucine production is 18 g / L.

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Abstract

It is an L-leucine high-yield bacterium and the production of L-leucine with invoice method, belonging to biological engineering field. The invented bacterial is a corynebacterium glutamicum JH-27, using corynebacterium glutamicum ATCC13032 as starting strain, mutagenizing separately with diethyl sulfate, nitrosoguanidine and ultraviolet, adding separately analogue of branched chain amino acid into minimum medium and sulfaguanidine into complete medium and breeding to prepare, being an L-leucine high-yield bacterium resistant sulfaguanidine and branched chain amino acid, compared with starting strain, producing L-leucine with the strain and invoice method, it expresses the ability of accumulation of highly qualified L-leucine. Shaking flask and culturing JH-27 for 60 hours, the yield of L-leucine is 30-32 g / l. Fermenting in fermenter of 5L for 55 hours, the yield of L-leucine is 40g / l.

Description

technical field [0001] An L-leucine high-yielding bacterium and its fermentation method to produce L-leucine, relating to an L-leucine high-yielding bacterium resistant to sulfaguanidine and branched-chain amino acid structural analogues, its breeding method and its use The invention relates to producing L-leucine by bacterial strain fermentation, which belongs to the technical field of bioengineering. Background technique [0002] L-leucine is one of the 18 common amino acids and one of the 8 essential amino acids for the human body. It has important uses in medicine, food, feed, cosmetics and other industries. At present domestic L-leucine production method is mainly based on proteolytic extraction method, as the technology disclosed in CN1056830C, it adopts hair hydrolysis to prepare cystine, and then prepares L-leucine from waste liquid. Due to the complex amino acid composition of natural protein raw materials, the extraction process is complicated, the pollution is se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
Inventor 蔡立明宁健飞
Owner WUXI JINGHAI AMINO ACID
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