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Biological resolution method for amino acid

A technology of amino acids and new methods, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as the inability to directly split DL-N-acetylated amino acid esters

Inactive Publication Date: 2008-05-28
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reported bioresolution methods cannot directly resolve DL-N-acetylated amino acid esters

Method used

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  • Biological resolution method for amino acid

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Effect test

Embodiment 1

[0047] The resolution of embodiment 1 L-phenylalanine

[0048] 1) Preparation of biocatalyst:

[0049] In a 500mL Erlenmeyer flask, Aspergillus Oyzae Lb3085 was inserted. The formula of the selected medium is 2% glucose, 35% soybean milk, 2.3% corn steep liquor, 0.1% magnesium sulfate, 0.2% dipotassium hydrogen phosphate, pH is 6.8, fermented on a shaking table (280r / min) at a temperature of 34°C for 37h . The wet cells were filtered and stored in freezer.

[0050] 2) split hydrolysis

[0051] Add 200mL of deionized water, 18g of wet bacteria and 2mL of 0.05mol / L MnCl2 solution into a 500mL flask, and then add 10mL of 1,4-dioxane dissolved in 20mmol of DL-N-acetylphenylalanine methyl ester solution, mix well. React at 35°C for 10 h on a shaker with a rotation speed of 200 r / min. During the reaction, the pH was controlled at about 7.5.

[0052] Post-treatment, the bacterial cells were removed by filtration, the pH of the aqueous solution was adjusted to 4, and extracted ...

Embodiment 2

[0053] The preparation of embodiment 2 D-phenylalanine

[0054] The ethyl acetate solution obtained in the above example was concentrated to recover the solvent. The residue was dissolved in 10 mL of methanol and 20 mL of 2M hydrochloric acid solution, heated to 68° C., and reacted for 2 h. Adjust the pH to 4.5, concentrate and recover methanol. The aqueous solution was treated with the same resin method as in 3 to obtain D-phenylalanine with a yield of 40%.

Embodiment 3

[0055] The resolution of embodiment 3 L-p-fluorophenylalanine

[0056] The preparation of the biocatalyst is the same as in Example 1.

[0057] 200mL of deionized water, 16g of wet bacteria, 2mL of 0.05mol / LMnCl solution was added to the 500mL Erlenmeyer flask filled with 20mL of 1,4-dioxane solution of 10mmol of N-acetyl-p-fluorophenylalanine methyl ester, Mix well. Under the condition of 28° C., the reaction was carried out on a shaker with a rotation speed of 200 r / min for 13 hours. During the reaction, the pH was adjusted to maintain around 7.2.

[0058] Aftertreatment is the same as in Example 1, and the yield is 41%.

[0059] The preparation of D-p-fluorophenylalanine is the same as in Example 2.

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Abstract

The invention relates to the field of biochemical engineering, in particular to a novel process for preparing L-amino acid and D-amino acid by direct biocatalysis and hydrolysis resolution for DL-N-acetyl amino acid esters. DL-N-acetyl amino acid esters is used as substrate, enzyme including amide hydrolase and ester hydrolase is used as catalyst, wherein the enzyme is achieved by fermenting aspergillus oryzae Lb3085, de-ionized water is utilized as solvent and added in an enzyme catalysis reactor, and L-N-acetyl amino acid esters of the substrate is hydrolyzed into L-amino acid, D-N-acetyl amino acid esters of the substrate is hydrolyzed into D-N-acetyl amino acid, thereby resolution is realized. The resolution technique of 'one bacterium two enzymes' can efficiently reduce operating intermediate process, and achieve purposes of convenience and cleaning.

Description

technical field [0001] The invention relates to the fields of biochemical industry and the like, in particular to a new method for preparing L-amino acid and D-amino acid by direct biocatalytic hydrolysis and splitting of DL-N-acetylamino acid ester. Background technique [0002] There are mainly 20 kinds of amino acids that make up organisms and participate in biological metabolism. The rest of the amino acids are called unnatural amino acids, including D-configuration amino acids, amino acids formed by adding or replacing different groups in the side chain with natural amino acids as the parent. and β or γ-amino acids of three types. In recent years, with the discovery of more and more biologically active peptides, the demand for amino acids, especially amino acids with special structures, is increasing, and its application and development have become a very active research field. [0003] Most amino acids have chiral structures, either D-form or L-form. However, amino a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P41/00C12P13/04C12N1/14C12R1/69
Inventor 刘守信甄小丽纪德彬康怀萍胡素坤丁文科
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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