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Method for producing L-2-aminobutyric acid by virtue of biological catalysis

An aminobutyric acid and amino acid technology, applied in the field of biosynthesis, can solve the problems of difficult separation and purification, harsh reaction conditions, cumbersome processes, etc., and achieve the effects of improving chiral purity, low equipment requirements, and simple steps.

Active Publication Date: 2015-07-15
HUNAN FLAG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at a series of defects such as cumbersome process, harsh reaction conditions, low yield, difficult separation and purification, and serious environmental pollution in the synthesis method of L-alpha-aminobutyric acid in the prior art, the purpose of the present invention is to provide a low-cost The method for synthesizing high-purity L-α-aminobutyric acid in a "one-pot method" using four kinds of enzyme complexes as catalysts as raw materials, the applicant's creative discovery, using specific ratios and types of enzymes, can prepare L-α-aminobutyric acid with high purity and yield, the method uses water phase as the reaction system, environmental protection, cheap, low equipment requirements, enzyme can be used repeatedly, low cost, easy large-scale industrial production

Method used

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  • Method for producing L-2-aminobutyric acid by virtue of biological catalysis
  • Method for producing L-2-aminobutyric acid by virtue of biological catalysis
  • Method for producing L-2-aminobutyric acid by virtue of biological catalysis

Examples

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example 1

[0041]Take 75.0g of anhydrous glycine and add an appropriate amount of deionized water to dissolve it, then add 120mL of absolute ethanol solution and dissolve it with 3.0mol / L ammonia water, adjust the pH to 8.0±0.05 with hydrochloric acid, remove insoluble matter by vacuum filtration, and set the volume to 1000mL. Add liquid alcohol dehydrogenase 4500U, immobilized threonine aldolase 3000U, immobilized threonine deaminase 6000U and immobilized L-amino acid dehydrogenase 4500U, NAD28.0mg, pyridoxal phosphate 15.0mg, control The reaction temperature is 35°C, and the pH is maintained at 8.00±0.05 with 3.0 mol / L ammonia water. When the molar conversion rate of glycine detected by the liquid phase is ≥99%, the conversion termination solution is filtered out. The stop solution was adjusted to pH 6.0±0.05 with 5.0 mol / L formic acid, and then electrodialysis was performed to desalt and remove water to obtain 1530 mL of a solution containing 92.04 g of L-α-aminobutyric acid, with a yi...

example 2

[0043] Take 75.0g of anhydrous glycine and add an appropriate amount of deionized water to dissolve it, then add 120mL of absolute ethanol solution and dissolve it with 3.0mol / L ammonia water, adjust the pH to 8.0±0.05 with hydrochloric acid, remove insoluble matter by vacuum filtration, and set the volume to 1000mL. Add liquid alcohol dehydrogenase 25000U, immobilized threonine aldolase 5000, immobilized threonine deaminase 25000 and immobilized L-amino acid dehydrogenase 20000U, NAD28.0mg, pyridoxal phosphate 15.0mg, control The reaction temperature is 35°C, and the pH is maintained at 8.00±0.05 with 3.0 mol / L ammonia water. When the molar conversion rate of glycine detected by the liquid phase is ≥99%, the conversion termination solution is filtered out. The stop solution was adjusted to pH 6.0±0.05 with 5.0 mol / L formic acid, and then electrodialysis was performed to desalt and remove water to obtain 1530 mL of a solution containing 95.04 g of L-α-aminobutyric acid, with a ...

example 3

[0045] Take 75.0g of anhydrous glycine and add an appropriate amount of deionized water to dissolve it, then add 120mL of absolute ethanol solution and dissolve it with 3.0mol / L ammonia water, adjust the pH to 8.0±0.05 with hydrochloric acid, remove insoluble matter by vacuum filtration, and set the volume to 1000mL. Add liquid alcohol dehydrogenase 8000U, immobilized threonine aldolase 4000U, immobilized threonine deaminase 12000U / mol and immobilized L-amino acid dehydrogenase 8000U, NAD28.0mg, pyridoxal phosphate 15.0mg , control the reaction temperature at 35° C., use 3.0 mol / L ammonia water to maintain the pH at 8.00±0.05, and filter out the conversion termination solution when the molar conversion rate of glycine is ≥99% as detected by the liquid phase. The stop solution was adjusted to pH 6.0±0.05 with 5.0 mol / L formic acid, and then electrodialysis was performed to desalt and remove water to obtain 1530 mL of a solution containing 95.44 g of L-α-aminobutyric acid, with a...

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Abstract

The invention discloses a new method for synthesising L-alpha-aminobutyric acid with a high optical activity. The new method comprehensively applies alcohol dehydrogenase, threonine aldolase, threonine deaminase and L-amino acid dehydrogenase, and ethanol and glycine raw material are directly synthesised into L-alpha-aminobutyric acid by a 'one-pot' method. According to the method disclosed by the invention, L-alpha-aminobutyric acid is synthesised in one pot by virtue of the simple and easily available raw materials; the method has the advantages of being few in operation steps, environment-friendly, good in bio-safety and simple in equipment, and is beneficial to industrial large-scale production for L-alpha-aminobutyric acid with a high optical activity and L-alpha-aminobutyric acid series products.

Description

technical field [0001] The invention relates to a method for synthesizing L-alpha-aminobutyric acid with a novel biocatalytic "one-pot method" and purifying, concentrating and crystallizing to obtain high-purity L-alpha-aminobutyric acid, which belongs to the field of biosynthesis. Background technique [0002] L-α-aminobutyric acid is an unnatural amino acid that inhibits the transmission of human nerve information. It has the effect of enhancing the activity of glucose phosphatase and promoting the metabolism of brain cells; L-α-aminobutyric acid is also an important raw material for pharmaceutical intermediates. It is widely used in the synthesis of drugs, such as the anti-tuberculosis drug ethambutol hydrochloride, the anti-epileptic drug levetiracetam and other drugs. [0003] At present, the production methods of L-α-aminobutyric acid at home and abroad are divided into chemical method and biological method. The chemical method is a relatively classic method, mainly in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/04
Inventor 许岗曾红宇赵士敏陈梅
Owner HUNAN FLAG BIOTECHNOLOGY CO LTD
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