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Method for separating and purifying L-leucine

A technology for separation and purification of leucine, applied in chemical instruments and methods, organic chemistry, cyanide reaction preparation, etc., can solve the problem of not separating L-leucine and L-valine, reducing L-leucine Acid yield and other issues, to avoid excessive losses, increase difficulty, and improve efficiency

Active Publication Date: 2019-05-17
内蒙古拜克生物有限公司
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AI Technical Summary

Problems solved by technology

This method adopts the reverse micelles extraction method. Although the extraction rate has been greatly improved, it still does not effectively separate L-leucine and L-valine.
Yu Wei et al. ("Separation and Extraction of L-Leucine in Fermentation Broth by Ion Exchange Method", Chemical World, 2005 (2): 109-113) based on the comparison of the solubility of L-leucine and L-valine in water Great characteristics, during the pretreatment of the fermentation broth, adopt the method of adding L-leucine seed crystals and twice recrystallization to remove most of the impurity amino acids, and then use 732 cation exchange resin to separate and purify L-leucine to Ammonium chloride was used as eluent to carry out gradient elution, and L-leucine was successfully separated from L-valine. Although this method separated L-leucine and L-valine, it was greatly Reduced yield of L-leucine

Method used

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  • Method for separating and purifying L-leucine
  • Method for separating and purifying L-leucine

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Experimental program
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Embodiment 1

[0029] Embodiment 1 A kind of separation and purification method of L-leucine

[0030] (1) add 20g polyaluminum iron and 30mg chitosan to every liter of fermentation broth, let stand, and centrifuge to obtain supernatant for later use;

[0031] (2) The pH value of the supernatant was adjusted to 1.5, sodium chloride was added to make the sodium chloride concentration reach 0.25g / ml, then L-leucine seed crystals were added, and the precipitate was separated by standing at 4°C for 24 hours;

[0032] (3) Add 15 times of water at 85°C to dissolve the precipitate, add 4% of the solution volume of activated carbon to decolorize for 1 hour, after centrifugation, adjust the pH of the supernatant to 5.5, concentrate to 1 / 10 of the original volume, let stand, and precipitate out crystal;

[0033] (4) The crystals were dissolved in water, passed through 007×7 strong cation exchange resin chromatography, eluted with 1 mol / L ammonia water, and the flow rate was controlled to be 2 ml / min. ...

Embodiment 2

[0034] Embodiment 2 A kind of separation and purification method of L-leucine

[0035] (1) add 15g zinc sulfate and 40mg diatomaceous earth in every liter of fermented liquid, let stand, centrifuge to obtain supernatant for subsequent use;

[0036] (2) The pH value of the supernatant was adjusted to 1.0, and sodium chloride was added to make the sodium chloride concentration reach 0.20 g / ml, then L-leucine seed crystals were added, and the precipitate was separated by standing at 4°C for 24 hours;

[0037] (3) Add 10 times water at 90°C to dissolve the precipitate, add 2% of the solution volume of activated carbon to decolorize for 1 hour, and after centrifugation, adjust the pH of the supernatant to 5.0, concentrate to 1 / 8 of the original volume, let stand, and precipitate out crystal;

[0038] (4) The crystals were dissolved in water, chromatographed through D001 cation exchange resin, eluted with 0.5 mol / L ammonia water, and the flow rate was controlled to be 1 ml / min. The...

Embodiment 3

[0039] Embodiment 3 A kind of separation and purification method of L-leucine

[0040] (1) add 22g zinc sulfate and 25mg polyacrylamide in every liter of fermentation broth, let stand, centrifuge to obtain supernatant for subsequent use;

[0041] (2) The pH value of the supernatant was adjusted to 2.0, and sodium chloride was added to make the sodium chloride concentration reach 0.30 g / ml, then L-leucine seed crystals were added, and the precipitate was separated by standing at 4°C for 24 hours;

[0042] (3) Add 20 times of water at 80°C to dissolve the precipitate, add 5% of the solution volume of activated carbon to decolorize for 1.5h, after centrifugation, adjust the pH of the supernatant to 6.0, concentrate to 1 / 15 of the original volume, let stand, Precipitation of crystals;

[0043] (4) The crystals are dissolved in water, chromatographed by D113 cation exchange resin, eluted with 2 mol / L ammonia water, and the flow rate is controlled to be 3 ml / min. The eluate is coll...

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Abstract

The invention provides a method for separating and purifying L-leucine. The method comprises the following steps: (1) adding a coagulant and a flocculant into fermentation broth, and reserving supernatant; (2) adding sodium chloride and a L-leucine seed crystal into the supernatant to separate precipitate; (3) adding water to dissolve the precipitate and decolorizing; (4) adding water to dissolvethe decolorized crystal, and performing ion exchange resin chromatography purification. According to the method, by the interaction of the coagulant and the flocculant in, bacteria, proteins, lipids and other substances in the fermentation broth are effectively removed, and the separation efficiency is greatly improved; on the other hand, by adding the sodium chloride and the L-leucine seed crystal at the same time, the solubility of the L-leucine is changed, the separation efficiency of the L-leucine and L-valine is improved, and the purity of the L-leucine is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for separating and purifying leucine. Background technique [0002] L-leucine, also known as leucine, is one of the essential amino acids for humans and animals. In 1819, Proust first isolated L-leucine from cheese, and later Bracon-not obtained its crystallization from the acid hydrolyzate of muscle and wool, and named it leucine. There are only 20 kinds of amino acids that constitute human protein, of which 8 are essential amino acids. From a nutritional point of view, evaluating the nutritional value of a protein mainly depends on the content and types of these 8 essential amino acids. L-leucine accounts for the highest share of complete proteins in nutrition, and is widely used in many industries such as medicine, food, flavoring, animal feed and cosmetics. [0003] In the field of medicine, L-leucine is an important raw material for clinical compound crystalline amino a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C229/08C07C227/42
Inventor 宋春慧陈峰郑建勋刘培马光辉
Owner 内蒙古拜克生物有限公司
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