Method for extracting high-abundance L-arginine-15N4 from high-abundance 15N isotope-labeled L-arginine fermentation liquor

A technology of isotope labeling and -15N4, which is applied in organic chemical methods, chemical instruments and methods, and the preparation of organic compounds. It can solve the problems of long extraction time and achieve high column efficiency and large sample loading.

Inactive Publication Date: 2016-07-06
SHANGHAI RES INST OF CHEM IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method not only overcomes the long extraction time of the ion exchange resin method, but also avoids the toxic operation problem caused by

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  • Method for extracting high-abundance L-arginine-15N4 from high-abundance 15N isotope-labeled L-arginine fermentation liquor

Examples

Experimental program
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Effect test

Embodiment 1

[0026] L-Arginine- 15 N 4 Fermentation broth 100ml, L-Arg- 15 N 4 Content 1.53% (converted into L-Arg.HCl- 15 N 4 1.84%), adjust the pH value of the solution with 2mol / L hydrochloric acid to be 3, 4000rpm centrifugal precipitation mycelia, get the supernatant, load the sample by constant flow pump A, sample loading speed 5ml / min, chromatographic column volume 200ml, 20 minutes After loading the sample, wash the chromatographic column with distilled water at a flow rate of 5ml / min. After 180 minutes, the washing solution is basically colorless and the absorbance Au value of the detector does not decrease. Use 0.5mol / L ammonia water to wash the column. The flow pump B is sent in, and the fraction of each peak is collected through the indication of the detector, and analyzed by paper chromatography or mass spectrometry, and the fraction of the last peak is L-Arg- 15 N 4 Crude products, others containing L-Arg- 15 N 4 and other miscellaneous acids can be re-loaded to the c...

Embodiment 2

[0030] L-Arginine- 15 N 4 Fermentation broth 500ml, L-Arg- 15 N 4 Content 1.47% (converted into L-Arg.HCl- 15 N 4 1.76%), adjust the pH value of the solution with 3mol / L hydrochloric acid to be 2, centrifuge and precipitate mycelia at 5000rpm, get the supernatant, load the sample by constant flow pump A, the sample loading speed is 18ml / min, and the chromatographic column volume is 1000ml for 25 minutes After loading the sample, wash the chromatographic column with distilled water at a flow rate of 20ml / min. After 200 minutes, the washing solution is basically colorless and the absorbance Au value of the detector does not decrease. Use 0.8mol / L ammonia water to wash the column. The flow pump B is sent in, and the fraction of each peak is collected through the indication of the detector, and analyzed by paper chromatography or mass spectrometry, and the fraction of the last peak is L-Arg- 15 N 4 Crude products, others containing L-Arg- 15 N 4 and other miscellaneous aci...

Embodiment 3

[0034] L-Arginine- 15 N 4 Fermentation broth 500ml, L-Arg- 15 N 4 Content 1.47% (converted into L-Arg.HCl- 15 N 4 1.76%), adjust the pH value of the solution with 4mol / L hydrochloric acid to be 4, centrifuge the mycelia at 4000pm, get the supernatant, load the sample by constant flow pump A, the sample loading speed is 18ml / min, and the chromatographic column volume is 1000ml for 25 minutes After loading the sample, wash the chromatographic column with distilled water at a flow rate of 20ml / min. After 200 minutes, the washing solution is basically colorless and the absorbance Au value of the detector does not decrease. Use 0.8mol / L ammonia water to wash the column. The flow pump B is sent in, and the fraction of each peak is collected through the indication of the detector, and analyzed by paper chromatography or mass spectrometry, and the fraction of the last peak is L-Arg- 15 N 4 Crude products, others containing L-Arg- 15 N 4 and other miscellaneous acids can be put...

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Abstract

The present invention relates to high-abundance 15 Extraction of high-abundance L-arginine from N-isotope-labeled L-arginine fermentation broth 15 N 4 approach to high-abundance 15 The fermentation broth of N isotope-labeled L-arginine is used as raw material, and a medium-pressure preparative chromatography system is used. After two-step separation, pure L-arginine can be obtained after 8-12 hours. 15 N 4 The single-product solution is then concentrated by catching ammonia, decolorized by activated carbon, crystallized by absolute ethanol, and vacuum-dried at 60°C to separate and extract L-arginine- with a product purity > 98%. 15 N 4 product, the total extraction yield is greater than 80%. The invention not only has simple principle, convenient operation, large and fast processing capacity, but also avoids the long extraction time of the ion exchange resin method and the environmental pollution caused by the discharge of a large amount of waste acid and alkali produced by resin treatment.

Description

technical field [0001] The present invention relates to the technical field of producing amino acids by fermentation, in particular to a method for producing high-abundance L-arginine- 15 N 4 Rapid separation and extraction of L-arginine- 15 N 4 Methods. Background technique [0002] The chemical name of L-arginine (L-arginine) is α-amino-δ-guanidinovaleric acid, and its molecular formula is C 6 h 14 N 4 0 2 , with a molecular mass of 174.2, usually white orthorhombic (dihydrate) crystals or white crystalline powder, easily soluble in water, insoluble in ether, slightly soluble in ethanol. L-arginine is a semi-essential amino acid in humans and animals. It is the amino acid with the most functions in animal cells found so far. It is not only an important raw material for protein synthesis, but also creatine, polyamine and nitric oxide ( It is a synthetic precursor of many active substances such as NO) and has a variety of unique physiological and pharmacological effe...

Claims

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Application Information

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IPC IPC(8): C07C277/08C07C279/14
CPCC07C277/08C07B2200/05
Inventor 任征侯捷岳韩笑侯静华张亮刘占峰雷雯宋明鸣
Owner SHANGHAI RES INST OF CHEM IND
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