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Biosynthetic method of high purity L-alpha-amino acid

A biosynthesis and amino acid technology, applied in the direction of fermentation, can solve the problems of harsh reaction conditions, difficult separation and purification, and complicated process, and achieve the effects of low equipment requirements, wide source of raw materials, and simple method steps.

Active Publication Date: 2015-01-28
HUNAN BAOLISHI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at a series of defects such as cumbersome process, harsh reaction conditions, low yield, and difficult separation and purification in the synthesis method of L-leucine in the prior art, the purpose of the present invention is to provide a catalyst based on biological enzyme, which uses aldehyde A method for synthesizing high-purity L-alpha-amino acids with high yields under mild conditions with quasi-compounds and glycine raw materials. The method uses the water phase as the reaction system, is environmentally friendly, cheap, has low equipment requirements, and the enzyme can be used repeatedly. The cost is low and meets industrialization requirements. Production

Method used

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  • Biosynthetic method of high purity L-alpha-amino acid
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Examples

Experimental program
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Effect test

Embodiment 1

[0041] Take L-norvaline as an example:

[0042] Take 63.0g of anhydrous glycine and add an appropriate amount of deionized water to dissolve it, then add 97% 60mL propionaldehyde solution and dissolve it with 3.0mol / L ammonia water to adjust the pH to 7.50±0.05, vacuum filter to remove insoluble matter, and set the volume to 1000mL. Cast immobilized L-threonine aldolase 37500U / mol (propionaldehyde), immobilized L-threonine deaminase 25000U / mol (propionaldehyde), control reaction temperature 30°C, use 3.0mol / L ammonia water Keep the pH at 7.50±0.05, and when the molar conversion rate of glycine in the liquid phase is ≥99%, filter out the conversion termination solution. After separation and purification by ion exchange chromatography, 1156 mL of a solution containing 88.44 g of 2-pentanuonic acid was obtained, with a yield of 95.30%. The chromatographic collection liquid was then adjusted to pH 7.50±0.05 with 3.0mol / L ammonia water, and then immobilized leucine dehydrogenase w...

Embodiment 2

[0053] Take 63.0g of anhydrous glycine and add an appropriate amount of deionized water to dissolve it, then add 97% 60mL propionaldehyde solution and dissolve it with 3.0mol / L ammonia water to adjust the pH to 7.50±0.05, vacuum filter to remove insoluble matter, and set the volume to 1000mL. Cast immobilized L-threonine aldolase 37500U / mol (propionaldehyde), immobilized L-threonine deaminase 25000U / mol (propionaldehyde), control reaction temperature 30°C, use 3.0mol / L ammonia water Keep the pH at 7.50±0.05, when the molar conversion rate of glycine in the liquid phase is ≥99%, filter out the conversion termination solution. After separation and purification by ion exchange chromatography, 1156 mL of a solution containing 88.44 g of 2-pentanuonic acid was obtained, with a yield of 95.30%. The chromatographic collection liquid was then adjusted to pH 7.50±0.05 with 3.0mol / L ammonia water, and immobilized leucine dehydrogenase was added at a ratio of 12000U / mol (2-pentanuonic ac...

Embodiment 3

[0055] Take 63.0g of anhydrous glycine and add an appropriate amount of deionized water to dissolve it, then add 97% 60mL propionaldehyde solution and dissolve it with 3.0mol / L ammonia water to adjust the pH to 7.50±0.05, vacuum filter to remove insoluble matter, and set the volume to 1000mL. Cast immobilized L-threonine aldolase 37500U / mol (propionaldehyde), immobilized L-threonine deaminase 25000U / mol (propionaldehyde), control reaction temperature 30°C, use 3.0mol / L ammonia water Keep the pH at 7.50±0.05, and when the molar conversion rate of glycine in the liquid phase is ≥99%, filter out the conversion termination solution. After separation and purification by ion exchange chromatography, 1156 mL of a solution containing 88.44 g of 2-pentanuonic acid was obtained, with a yield of 95.3%. The chromatographic collection liquid was then adjusted to pH 7.50±0.05 with 3.0mol / L ammonia water, and immobilized leucine dehydrogenase was added at a ratio of 12000U / mol (2-pentanuonic...

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Abstract

The invention discloses a biosynthetic method of high purity L-alpha-amino acid. The method comprises the following steps: in a reaction liquid, carrying out enzymic catalytic reaction on an aldehyde compound and a glycine raw material under the catalytic effect of L-threonine aldolase and L-threonine dehydratase compound enzymes to obtain a 2-ketonic acid product; and carrying out enzymic catalytic reaction on the 2-ketonic acid product under catalysis of a coenzyme consisting of leucine dehydrogenase and hydrogenlyase or glucose dehydrogenase, and sequentially carrying out electrodialysis desalting, active carbon decoloring, concentration and crystallization and vacuum drying on the reaction product so as to obtain high purity L-alpha-amino acid crystals with a high yield. The method which uses an aqueous phase as a reaction system is environment-friendly and cheap, low in equipment requirement and low in cost, and the enzyme can be repeatedly used, so that the method can be used for satisfying industrial production.

Description

technical field [0001] The invention relates to a method for converting and synthesizing L-α-amino acid through a novel biological method and obtaining high-purity L-α-amino acid through purification, concentration and crystallization, which belongs to the field of biosynthesis. Background technique [0002] Amino acids are the basic substances that make up proteins and are necessary for human life activities. They are responsible for the synthesis of muscles, skin, organs, enzymes, and immune antibodies in the body. Generally speaking, amino acids are divided into D-type and L-type, and only L-type amino acids in the human body participate in metabolism and have physiological activity. [0003] L-norvaline is a key intermediate in the synthesis of perindopril, ACE-inhibitors - antihypertensive drugs. L-norvaline can be obtained by splitting racemic DL-norvaline, or by direct synthesis, and there are few related reports. Japanese patent JP7553587 is produced by fermentatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/04
Inventor 许岗曾红宇帅得利王胜锋
Owner HUNAN BAOLISHI BIOTECH
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