CN1515543A

CN1515543A - Method for separating 14 amino acids by utilizing active carbon adsorption column and adopting once-through method

Info

Publication number: CN1515543A
Application number: CNA021349231A
Authority: CN
Inventors: 邹学满
Original assignee: Individual
Current assignee: Individual
Priority date: 2002-10-10
Filing date: 2002-10-10
Publication date: 2004-07-28

Abstract

The method for separating 14 amino acids by means of active carbon adsorption column one-step operation includes the following steps: diluting animal and plant protein hydrolysate and regulating pH value, making it flow through active carbon adsorption column, using water, acid liquor and ethyl alcohol to prepare eluent, continuously stage collecting, mixing the amino acids of alanine and leucine, etc. with eluent and making them flow through anion-exchange column, and making the eluent mixed with arginine flow through cation-exchange column, then fine separating out single amino acid. Said invention uses human hair as protein hydrolysis raw material, and uses said invented process to make chromatographic separation so as to can obtain 14 single amino acids, its total yield rate is 52.1%.

Description

The method of disposable separation 14 seed amino acids of active carbon adsorption column

The present invention relates to amino acid separation from protein hydrolyzate, particularly high recovery rate ground separates some and is difficult to isolating amino acid whose method.

At present, amino acid separation is still continued to use traditional method from protein hydrolyzate, because the amino acid whose physico-chemical property of most of kinds is approximate, in addition identical, they can't be separated one by one.Generally be to utilize the relative quantity of certain seed amino acid in 17 seed amino acid mixed solutions big, the characteristics that solubleness is little are just decontroled it from mixed solution; Or utilize certain amino acid molecular to separate with the chemical substance complex-precipitation of certain special construction.Such separating resulting, the amino acid kind of acquisition is few, and recovery rate is low.The amino acid that is mixed in the acidic aqueous solution is wide in variety, and quantity is big, then can't utilize, and causes serious waste, and discharging causes environmental pollution again.Although CN85102242A patent application " protein hydrolysate separation method of amino-acids ", be that protein hydrolyzate directly is separated into A, B two portions by active carbon adsorption column earlier, the A part is separated into nine kinds of single amino acid kinds through resin column more at last after freeing; B part still has eight seed amino acids not to be separated, discharging and having wasted.

The objective of the invention is to adopt new technical matters, further improve the kind number of amino acid recovery rate and increase single amino acid.

The present invention and CN85102242A prior art are relatively, increased substantially this kind stratography technical matters, it is characterized in that protein hydrolyzate after dilution and regulating potential of hydrogen, the disposable active carbon adsorption column of flowing through, then free through washing, acidic solution and alkaline ethanol solution, contiguous segmentation is collected.Obtain third component, dried meat component, bright component, arginine solution in succession.In order to method processing down, at last each component amino acid is separated into single amino acid solution one by one again:

With third component and the dried meat component amino acid solution 717 resin anion(R.A) exchange columns of flowing through respectively, isolate the 1. amino acid of Radix Asparagi in succession; 2. L-glutamic acid; 3. Serine; 4. Threonine; 5. glycine; 6. Xie Ansuan; 7. proline(Pro); 8. Methionin; 9. nine kinds of single amino acids such as L-Ala.

Bright component is settled out amino acid mixing liquid behind the 1. leucine 717 resin anion(R.A) exchange columns of flowing through again with o-Xylol-4-sulfonic acid, isolates 2. methionine(Met) in succession; 3. Isoleucine; 4. Histidine.

Advantage of the present invention is that the amino acid in the protein hydrolyzate has been carried out full sepn process, make the amino acid kind number that is separated on nine kinds of bases of number of patent application 85102242, be increased to 14 kinds, total yield is increased to 52.1% from 41%, so, good separating effect of the present invention, the recovery rate height.

The present invention is described in further detail below in conjunction with example:

Send out and place 800 milliliters of round-bottomed flasks with 100 gram people; add 250 milliliters of hydrochloric acid; heat to more than 110 ℃; and hydrolysis 10 hours; collect hydrolysis then and transform the amino acid mixing liquid that generates; add the dilution of twice clear water; the liquefied ammonia pH value that neutralizes reaches more than 7.0; allow the active carbon adsorption column that the GH-15 granulated active carbon is housed in its  4.0cm * 20cm that flows through then; inject after the feed liquid, then, take off with hydrochloric acid and 50% left and right sides ethanol lyolysis of 6 to 8 times of charcoal column volume 0.05N-0.2N then with the clear water washing of twice charcoal column volume; the flow through solution of active carbon adsorption column of Fractional Collections obtains third component respectively; the dried meat component; bright component; arginic amino acid separation solution.Again third component and dried meat component are transferred to pH value more than 10, allow their flow through respectively  4.0cm * 20cm717 resin anion(R.A) exchange columns of regeneration H0-type, be separated to 1. aspartic acid; 2. L-glutamic acid; 3. Serine; 4. Threonine; 5. glycine; 6. Xie Ansuan; 7. proline(Pro); 8. Methionin; 9. the solution of the single amino acid of L-Ala, and then difference evaporation concentration obtains aspartic acid 4.1 grams, L-glutamic acid 9.9 grams, Serine 6.1 grams, Threonine 4.2 grams, glycine 2.5 grams, L-Ala 2.3 grams, Xie Ansuan 3.2 grams, proline-4 .8 gram, Methionin 1.8 grams; Be settled out 1. leucine 4.6 grams with o-Xylol 4-sulfonic acid; Remaining mixed solution transfers pH value allow the OH of its  2.0cm * 10.0cm that flows through more than 10 ^-Type 717 resin anion(R.A) exchange columns are isolated 1. methionine(Met) 0.4 gram, 2. Isoleucine 1.6 grams; Transfer pH value 8.0 allow the H of its  2.0cm * 10.0cm that flows through arginine solution ⁺Type 732 cation exchange resin columns obtain 1. arginine 6.2 grams, 2. Histidine 0.4 gram.More than 14 kinds of single amino acid total amounts be 52.1 grams, total recovery rate is 52.1%.

Document:

Prior art: Chinese invention patent application number 85102242

Claims

1, the method for protein hydrolysate amino acid separation is characterized in that 2 to 4 times of protein hydrolyzate thin ups, and with liquefied ammonia neutralize pH value more than 7.0 after, allow it directly enter active carbon adsorption column.Aminoacids solution is all flowed through behind the activated carbon column, with the washing of flowing through of 2 times of volume clear water of activated carbon column, then with the 0.05N-0.2N hydrochloric acid solution of 4 to the 6 times of activated carbon column volumes activated carbon column of flowing through, at last with the 0.2N hydrochloric acid solution of 1 times of activated carbon column volume and 50% left and right sides ethanol liquid activated carbon column of flowing through.All the flow through amino acid parting liquid of activated carbon column of Fractional Collections is judged third component, dried meat component, bright component, Histidine, arginine etc. by analysis.

1. allow third component and the dried meat component 717 resin anion(R.A) posts of flowing through respectively, separate obtaining nine kinds of single amino acids such as Serine, Threonine, L-Ala, glycine, aspartic acid, L-glutamic acid, Xie Ansuan, proline(Pro) and Methionin;

2. separate bright component liquid with o-Xylol-4-sulfonic acid complex-precipitation and obtain leucine, Isoleucine and methionine(Met).

Cutin protein such as 2, protein hydrolysate separation method of amino-acids according to claim 1, its characteristic protein matter are behaved and sent out, pig hair, or raise the protein of low values such as blood meal, gelatin, gelatine, residual silk, pupa albumen, plant oil cake.

3, the method for protein hydrolysate amino acid separation according to claim 1 is characterized in that it is chromatograph tripping devices such as active carbon adsorption column and 717 anion-exchange columns that the amino acid separation mixed solution becomes the equipment of single amino acid.

4, the method for protein hydrolysate amino acid separation according to claim 1 is characterized in that separating medium is gac, 717 resin anion(R.A)s, o-Xylol-materials such as 4-sulfonic acid.

5, the method for protein hydrolysate amino acid separation according to claim 1, its feature is freed the 70%-90% alcohol mixeding liquid that liquid is water, 0.05N-0.2N hydrochloric acid solution, 50% left and right sides alcoholic acid 0.2N hydrochloric acid mixed solution and 1N ammoniacal liquor as separation.

6,, it is characterized in that adopting the Fractional Collections method to collect the amino acid solution that is separated to according to the method for claim 1,3,4,5 described protein hydrolysate amino acid separations.