Method for synthesizing beta-alanine by biological catalysis

A technology of biocatalysis and alanine, applied in the field of enzyme engineering, can solve the problems of high price of β-aminopropionitrile, low reaction concentration, and poor economic benefits, and achieve safe and environmentally friendly production process, high substrate concentration, and low production cost. low effect

Inactive Publication Date: 2013-01-02
ANHUI HUAHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method substrate β-aminopropionitrile is expensive, and the reaction concent...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Determination of content

[0029] (1) Determination of fumaric acid content (high performance liquid chromatography)

[0030] The enzymatic reaction solution was centrifuged at 10,000r / min to pellet the cells, and the supernatant was taken and diluted 10 times. At the same time, a fumaric acid standard sample (2mg / ml) was prepared for HPLC analysis.

[0031] Column model: Alltech organic acid chromatography column No.88645, 250mm×4.6mm

[0032] Mobile phase: 25mmol / L KH 2 PO 4 , pH2.5

[0033] Injection volume: 20μl

[0034] Column temperature: 35°C

[0035] Gradient mode: constant flow 1.0ml / min

[0036] Detector: UV wavelength 210nm

[0037] Calculation method: external standard method

[0038] (2) Determination of β-alanine content (high performance liquid chromatography)

[0039] Take the extracted and dried β-alanine sample and β-alanine standard sample (2mg / ml) respectively, and perform HPLC analysis.

[0040] Derivation method: Accurately ...

Embodiment 2

[0048] Example 2: Biocatalytic synthesis of β-alanine

[0049] Seed medium: 5g fumaric acid, 10g corn steep liquor powder, 10g polypeptide protein, 10g monosodium glutamate, MgSO 4 ·7H 2 O 0.2g, K 2 HPO 4 0.15g, 0.4g NaCl, add water to prepare 1L solution, adjust pH to 7.0 with ammonia water, and steam sterilize at 121°C for 20min.

[0050] Fermentation medium: 20g fumaric acid, 40g corn steep liquor dry powder, 40g polypeptide protein, 20g L-aspartic acid, 0.8g betaine hydrochloride, MgSO 4 ·7H 2 O 0.8g, K 2 HPO 4 0.6g, 0.4g NaCl, add water to prepare 4L of solution, adjust pH to 7.0 with ammonia water, and steam sterilize at 121°C for 20min.

[0051] Substrate solution: 100 g of fumaric acid, adjust the pH to 7.5 with ammonia water, and dilute to 1 L.

[0052] Escherichia coli slant strains were inserted into the seed medium, cultured on a shaker at 37°C for 24 hours at 200 r / min. 10% of the inoculum was transferred to the fermentation medium, 200r / min, 37°C s...

Embodiment 3

[0053] Example 3: Biocatalytic synthesis of β-alanine

[0054] Seed medium: 10g fumaric acid, 15g dry powder of corn steep liquor, 5g polypeptide protein, 8g monosodium glutamate, MgSO 4 ·7H 2 O 0.3g, K 2 HPO 4 0.20g, 0.25g NaCl, add water to prepare 1L solution, adjust pH to 7.2 with ammonia water, and steam sterilize at 121°C for 20min.

[0055] Fermentation medium: 40g fumaric acid, 60g dry powder of corn steep liquor, 20g polypeptide protein, 40g L-aspartic acid, 1.0g betaine hydrochloride, MgSO 4 ·7H 2 O 1.0g, K 2 HPO 4 0.8g, NaCl 1.0g, add water to prepare 4L of solution, adjust the pH to 7.2 with ammonia water, and steam sterilize at 121°C for 20min.

[0056] Substrate solution: 80 g of fumaric acid, adjust the pH to 7.0 with ammonia water, and dilute to 1 L.

[0057] Escherichia coli slant strains were inserted into the seed medium, cultured on a shaker at 32°C for 20 h at 250 r / min. 10% of the inoculum was transferred to the fermentation medium, 2...

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PUM

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Abstract

The invention provides a method for synthesizing beta-alanine by biological catalysis, and belongs to the field of enzyme engineering. The method comprises the following steps: culturing escherichia coli microbe to obtain aspartase and L-aspartic acid-alpha-decarboxylase, mixing the enzyme-containing escherichia coli cells with an ammonium fumarate aqueous solution, performing an enzyme reaction to generate beta-alanine. The method of the invention is cheap and easily available in raw materials, simple in production process, safe, environment-friendly, mild in reaction condition, high in efficiency, low in production cost, and high in product quality, and has significant economic benefits and environmental protection benefits when compared with traditional methods.

Description

technical field [0001] The invention relates to a biocatalytic method for synthesizing β-alanine, which belongs to the field of enzyme engineering. Background technique [0002] β-alanine (English name beta-Alanine), also known as β-alanine, is a non-protein amino acid and the only β-type amino acid in nature. In 1972, Ross and Monroe reported in the degradation products of uracil Find. β-alanine is an important biochemical raw material, which has a wide range of application fields and market prospects in the fields of medicine, feed and food. At present, β-alanine is mainly used in the synthesis of pantothenic acid and calcium pantothenate, pantothenic acid is also known as vitamin B 5 , is a component of coenzyme A necessary for many metabolisms. [0003] According to current bibliography, the synthetic method of β-alanine mainly contains following several kinds: [0004] 1. Chemical synthesis method [0005] (1) Acrylic acid method (US 2376334 A, 1945.05.22; US 31050...

Claims

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Application Information

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IPC IPC(8): C12P13/06C12R1/19
Inventor 不公告发明人
Owner ANHUI HUAHENG BIOTECH
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