Glutathion purification and separation method

A glutathione, purification and separation technology, applied in the preparation methods of peptides, chemical instruments and methods, peptides, etc., can solve the problems of low recovery rate, unsuitable glutathione, mercury pollution, etc., achieve volume reduction, The effect of improving single-column separation efficiency, pH value and wide range of elution control conditions

Inactive Publication Date: 2007-07-11
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the preparation of glutathione by metal ion mercury affinity chromatography has the advantages of good selectivity, etc., there are disadvantages such as mercury pollution and low recovery rate, especially not suitable for the production of medicinal and edible glutathione

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] The styrene-based resin containing -SH group was pretreated and packed in a 1.0cm×40cm chromatographic column, and the chromatographic column was equilibrated with 0.2M disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution with a pH value of 7.0, Then load 30ml (0.7mgGSH / ml) of the GSH extract with a pH value of 7.0, wash with the same buffer, and elute with 0.2M sodium hydroxide to obtain an eluent containing only GSH. The elution yield can reach 92%, and the purity after crystallization can reach 99%.

Embodiment 2

[0016] will contain -S-S-C 5 h 4 The N-group styrene resin was pretreated and packed into a 3.5cm×40cm chromatographic column, equilibrated with 0.5M Tris-hydrochloric acid buffer solution with a pH value of 10.0, and then used GSH to adjust the pH value to 10.0 Load 300ml (0.7mgGSH / ml) of the extract, wash with the same buffer, and then elute with 0.2M hydrochloric acid to obtain an eluate containing only GSH, and the elution yield can reach 86%. The purity reaches 99% after crystallization.

Embodiment 3

[0018] Pack the styrene-based resin containing -SH group into a 7.0cm×40cm chromatographic column after pretreatment, equilibrate the chromatographic column with 0.1M acetic acid-sodium acetate buffer solution with a pH value of 4.0, and then adjust the pH value with Load 1300ml (0.7mgGSH / ml) of the GSH extract of 4.0, wash with the same buffer, and then elute with a gradient of 0.1M to 0.2M ammonia water to obtain an eluate containing only GSH. The rate can reach 84%.

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Abstract

The invention offers a Process for the separation and purification of glutathione, selecting separation medium contains-S-Rgroup or -S-S-R'group, preprocessing the medium which separates glutathione, stuffing to chromatography column, balancing chromatography column via buffer liquid which the pH value is 1. 0-10. 0, sampling by the glutathione extracting liquid which pH value is 1. 0-10. 0, eluting by alkali or acid after washing by one buffer liquid can get the eluent which only contains glutathione. Because the medium selective adsorbing glutathione high specifically, the glutathione can be separated effectively, achieving high purification from glutathione buffer liquid by one-step separation, the product which the purity is 99% can be getted after crystallization of glutathione.

Description

technical field [0001] The invention relates to a method for purifying and separating glutathione (GSH), in particular to separating glutathione by ion exchange chromatography, and belongs to the field of biochemical industry. Background technique [0002] Glutathione is an active tripeptide with important physiological functions. It is formed by condensation of glutamic acid, cysteine ​​and glycine through peptide bonds. Its chemical name is γ-L-glutamyl-L-cysteine Aminoacyl-glycine. The relative molecular mass of glutathione is 307.33, the melting point is 189-193°C (decomposition), the crystal is colorless, transparent and slender columnar, and the isoelectric point is 5.93. It is soluble in water, dilute alcohol, liquid ammonia and dimethylformamide, but insoluble in alcohol, ether and acetone. The solid glutathione is relatively stable, while the aqueous solution is easily oxidized in the air. The active sulfhydryl groups of two molecules of reduced glutathione (GSH)...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K5/08C07K1/18
Inventor 谭天伟丁轲
Owner BEIJING UNIV OF CHEM TECH
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