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Method for preparing beta-alanine by coupled enzymatic reaction

An alanine and coupling technology, applied in the biological field, can solve the problems of industrialized production and environmental protection, such as the difficulty of industrial production and environmental protection, harsh process conditions, and difficult separation of intermediate products, and achieve outstanding implementation effects, high catalytic rate and conversion rate, The effect of reducing acid and alkali consumption and equipment investment

Inactive Publication Date: 2013-09-25
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the β-aminopropionitrile method and the acrylonitrile method have cheap raw materials, but it is difficult to separate the intermediate product after hydrolysis, especially the separation of β-alanine and by-products is complicated, and a large amount of inorganic salts are generated during the hydrolysis process.
The process conditions of this method are demanding, and other side reactions are prone to occur, so it is difficult to industrialize production and environmental protection, and the production cost is high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Mix 100 ml Escherichia coli str. 1.5 g and 1000 ml of aspartase wet bacteria obtained by centrifugation of K-12 substr. MG1655 fermentation broth Corynebacterium glutamicum 17 g of aspartic acid-α-decarboxylase wet bacteria obtained by centrifugation of ATCC 13032 fermentation broth was added to 1000 ml of transformation liquid containing 150 g / L ammonium fumarate, 0.5 g / L OP, pH 7.0, 37 ℃ enzymatic reaction for 16 h, paper chromatography detection to determine the end of the reaction. After the reaction, the concentration of β-alanine in the conversion solution was 85.4 g / L, and the molar conversion rate to ammonium fumarate was 96%.

[0034] 2. Centrifuge the transformation solution at 4000 r / min for 15 minutes to remove the bacteria, heat the supernatant to 70°C-80°C, add activated carbon for decolorization, concentrate the decolorization solution to 120 ml, cool and crystallize at low temperature, filter with suction, wash, and dry. Obtain 49.6 g of solid β-...

Embodiment 2

[0037] 1. Mix 200 ml Escherichia coli str. 3.5 g and 1000 ml of aspartase wet bacteria obtained by centrifugation of K-12 substr. MG1655 fermentation broth Corynebacterium glutamicum 19 g of aspartic acid-α-decarboxylase wet bacteria obtained by centrifugation of ATCC 13032 fermentation broth was added to 1000 ml of transformation liquid containing 100 g / L ammonium fumarate, 0.05 g / L Tween- 80, pH 7.5, 37 ℃ enzymatic reaction for 12 h, paper chromatography detection to determine the reaction end point. After the reaction, the concentration of β-alanine in the conversion solution was 57.5 g / L, and the molar conversion rate to ammonium fumarate was 97%.

[0038] 2. Centrifuge the transformation solution at 4000 r / min for 15 minutes to remove the bacteria, heat the supernatant to 70°C-80°C, add activated carbon for decolorization, concentrate the decolorization solution to 80 ml, cool and crystallize at low temperature, suction filter, wash, and dry. Obtained 32.3 g of solid...

Embodiment 3

[0041] 1. Mix 100 ml Escherichia coli str. K-12 substr. MG1655 fermentation broth centrifuged 1.4 g of aspartase wet bacteria was added to 1000 ml of transformation liquid containing 200 g / L ammonium fumarate, 5 g / L Tween-80 , pH 7.0, 30℃ enzymatic reaction for 8 h, then add 1000 ml to the transformation solution Corynebacterium glutamicum 16 g of aspartic acid-α-decarboxylase wet bacteria obtained by centrifugation of ATCC 13032 fermentation broth continued to react for 20 h, and the end point of the reaction was determined by paper chromatography. After the reaction, the concentration of β-alanine in the conversion solution was 113.9 g / L, and the molar conversion rate to ammonium fumarate was 96%.

[0042] 2. Centrifuge the transformation solution at 4000 r / min for 15 minutes to remove bacteria, heat the supernatant to 70°C-80°C, add activated carbon for decolorization, concentrate the decolorization solution to 155 ml, cool and crystallize at low temperature, suction fil...

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Abstract

The invention belongs to the biotechnical field, and in particular relates to a method for preparing beta-alanine by a coupled enzymatic reaction. The method comprises the following steps: by using ammonium fumarate as a raw material, mixing bacterial cells respectively comprising aspartase and aspartic acid-alpah-decarboxylase or a crude enzyme liquor of the two enzymes with an ammonium fumarate aqueous liquor, wherein pH of the ammonium fumarate aqueous liquor is 7.0-7.5; performing enzymatic reaction at 25-55 DEG C; obtaining high purity beta-alanine by separating converted products by isoelectric point crystallization or a method combining isoelectric point crystallization or ion exchange resin. The beta-alanine is prepared by coupled enzymatic reaction, and the method has the advantages of wide source of raw materials, low cost, simplicity and convenience in operation, short enzymatic conversion time, low production cost and the like.

Description

1. Technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing beta-alanine by a double-enzyme method. 2. Background technology [0002] β-Alanine is a precursor for the synthesis of pantothenic acid, a B vitamin that forms part of coenzyme A and is important for endogenous metabolic energy exchange in various tissues. With the gradual deepening of the understanding of β-alanine, β-alanine is widely used in medicine, food, cosmetics, feed and other fields, and the market demand for β-alanine is also growing. Therefore, β-alanine Amino acid has broad application prospects and development value. [0003] At present, according to literature reports, the preparation methods of β-alanine mainly include chemical synthesis and biological enzymatic methods. [0004] 1. Chemical synthesis method [0005] Chemical synthesis methods include acrylonitrile method, acrylic acid method, succinimide (succinimide) degr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/06
Inventor 焦庆才刘均忠毛娉婷刘茜
Owner NANJING UNIV
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