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Comamonas testosteroni HHALA-001 and method for using comamonas testosteroni HHALA-001 to produce L-alanine

A technology of Comamonas and testosterone, applied in the field of bioengineering, can solve the problems of low L-alanine conversion rate and conversion efficiency, insufficient enzyme activity, etc., and achieves improved production efficiency and conversion rate, good stability, The effect of reducing production costs

Active Publication Date: 2017-10-20
ANHUI HUAHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, although there are bacterial strains that can convert L-aspartic acid, there are problems such as insufficient enzyme activity, low conversion rate and conversion efficiency of L-alanine, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Activated slant medium (g / L): peptone 8.0, sodium chloride 5.0, yeast extract (powder) 3.0, beef extract 5.0, agar 15, adjust pH 7.0-7.2.

[0044] Shake flask seed medium (g / L): sodium glutamate 15.0, compound nitrogen 10.0, corn steep liquor dry powder 6.0, magnesium sulfate heptahydrate 0.25, dipotassium hydrogen phosphate 0.2, potassium dihydrogen phosphate 0.2, fumaric acid 0.8, Ammonium sulfate 1.0, adjust pH 7.0-7.2 with sodium hydroxide solution.

[0045] Fermentation medium (g / L): compound nitrogen 10.0, yeast extract 1.0, corn steep liquor dry powder 4.0, sodium glutamate 12.0, magnesium sulfate heptahydrate 0.1, ammonium sulfate 0.8, citric acid monohydrate 2.0, dipotassium hydrogen phosphate 0.2 , potassium dihydrogen phosphate 0.5, fumaric acid 1.0, adjust the pH to 7.0-7.2 with sodium hydroxide solution.

[0046] Take one ring of Comamonas testosteroni HHALA-001 slant strain and connect it to the slant medium, culture at 30°C±1°C for 20h; then take one ring ...

Embodiment 2

[0048] Activated slant medium (g / L): peptone 14.0, sodium chloride 6.0, yeast extract (powder) 8.0, beef extract 8.0, agar 20, adjust pH 7.0-7.2.

[0049] Shake flask seed medium (g / L): sodium glutamate 5.0, compound nitrogen 5.0, corn steep liquor dry powder 10.0, magnesium sulfate heptahydrate 0.25, dipotassium hydrogen phosphate 0.7, potassium dihydrogen phosphate 0.7, fumaric acid 4.5, Ammonium sulfate 1.5, adjust the pH to 7.0-7.2 with sodium hydroxide solution.

[0050] Fermentation medium (g / L): compound nitrogen 14.0, yeast extract 2.5, corn steep liquor dry powder 14.0, sodium glutamate 27.0, magnesium sulfate heptahydrate 0.25, ammonium sulfate 1.6, citric acid monohydrate 0.7, dipotassium hydrogen phosphate 0.8 , potassium dihydrogen phosphate 0.8, fumaric acid 4.2, adjust the pH to 7.0-7.2 with sodium hydroxide solution.

[0051] Take one ring of Comamonas testosteroni HHALA-001 slant strain and connect it to the slant medium, culture at 30°C±1°C for 20h, then tak...

Embodiment 3

[0053] Activated slant medium (g / L): peptone 14.0, sodium chloride 6.0, yeast extract (powder) 8.0, beef extract 8.0, agar 20, adjust pH 7.0-7.2.

[0054] Shake flask seed medium (g / L): sodium glutamate 5.0, compound nitrogen 5.0, corn steep liquor dry powder 10.0, magnesium sulfate heptahydrate 0.25, dipotassium hydrogen phosphate 0.7, potassium dihydrogen phosphate 0.7, fumaric acid 4.5, Ammonium sulfate 1.5, adjust the pH to 7.0-7.2 with sodium hydroxide solution.

[0055] Fermentation medium (g / L): compound nitrogen 13.0, yeast extract 3.0, corn steep liquor dry powder 12.0, sodium glutamate 25.0, magnesium sulfate heptahydrate 0.25, ammonium sulfate 1.6, citric acid monohydrate 0.8, dipotassium hydrogen phosphate 0.8 , potassium dihydrogen phosphate 0.8, fumaric acid 3.5, adjust the pH to 7.0-7.2 with sodium hydroxide solution.

[0056] Take one ring of Comamonas testosteroni HHALA-001 slant strain and connect it to the slant medium, culture at 30°C±1°C for 20h, then tak...

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Abstract

The invention belongs to the technical field of biological engineering, and in particular relates to comamonas testosteroni HHALA-001 and a method for using the comamonas testosteroni HHALA-001 to produce L-alanine; the preservation number of the strain is CGMCCNo.14064; the comamonas testosteroni HHALA-001 is obtained by compound mutation and repeated screening, the strain has high enzyme activity, the enzyme activity can reach 11.80U-12.22U, the strain can be used for high expression of L-aspartate beta-decarboxylase, is used for production of the L-alanine by catalysis of L-aspartic acid, can greatly improve production efficiency and conversion rate of the L-alanine, helps to reduce production cost, and has good stability; after use of the strain, the conversion rate of the L-Aspartic acid can reach 99.9%; the comamonas testosteroni HHALA-001 fully meets industrial L-alanine strain mutagenesis and screening demands, can be used in industrial production, and has great economic value.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to Comamonas testosteroni HHALA-001 and a method for producing L-alanine by using the strain. Background technique [0002] L-alanine is one of the amino acids closely related to sugar metabolism in the living body. It has a wide range of uses in the fields of food and medicine, and has a large market demand. At present, the production methods of L-alanine generally include proteolytic extraction, fermentation and enzymatic conversion. Wherein, the enzymatic conversion method refers to catalyzing L-aspartic acid by microbial cells rich in L-aspartic acid-β-decarboxylase activity to obtain L-alanine. Because the enzymatic conversion method has the advantages of low production cost, simple production process, high product yield and purity, it is widely used in industry. In the prior art, although there are bacterial strains capable of converting L-aspartic acid, t...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P13/06C12R1/01
CPCC12P13/06C12N1/205C12R2001/01
Inventor 郭恒华张冬竹田宋魁章晖
Owner ANHUI HUAHENG BIOTECH
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