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Method for isolating and purifying branched-chain amino acid from fermented liquid

A technology for separation and purification of branched-chain amino acids, applied in the field of separation and purification of branched-chain amino acids, which can solve problems such as environmental pollution, high consumption, inability to separate miscellaneous acids, etc., to solve the problem of consumption and salt pollution, clean production, energy saving and emission reduction, and acid reduction The effect of alkali consumption

Inactive Publication Date: 2019-02-26
丁杰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Now, the production of branched-chain amino acids mainly relies on microbial fermentation, but other amino acids will be produced during microbial fermentation. The molecular weight and chemical isoelectric point of these amino acids, especially neutral amino acids (such as leuc amino acids), are very close to branched-chain amino acids. Caused problems such as low acid production level and high heteroacid content in the process of microbial fermentation to produce branched chain amino acids
At present, the ion exchange method is often used to extract branched-chain amino acids in the fermentation broth of microorganisms. However, the ion exchange process cannot separate heteroacids and has many defects. For example, the ion exchange process steps are complicated and require regeneration and elution; ion exchange A large amount of acid and alkali will be used in the regeneration process of the process, resulting in low product purity and low yield; the ion exchange process will produce a large amount of ammonium nitrogen waste liquid after elution and regeneration, which will not only consume a large amount of acid and alkali It costs a lot of money and will cause great pollution to the environment. At the same time, the elution process will consume a lot of water
Therefore, the backwardness of separation technology limits the large-scale production of branched-chain amino acids, and because other amino acids in branched-chain amino acid products are difficult to remove, domestic high-end API products cannot meet EU standards, so they cannot enter the international market to participate in the competition of APIs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0008] Example 1: The specific steps for the separation and purification of valine are as follows: (1) filter and sterilize the valine fermentation broth and concentrate it to 5 times, and then spray-dry the concentrated fermentation broth to obtain crude valine; (2) Mix the crude product of valine with hydrochloric acid according to the mass ratio of 1:3, and dissolve for the first time at 80° C. to obtain the crude solution of valine; (3) cool the crude solution of valine to Filter after 25°C to obtain valine hydrochloride; (4) add valine hydrochloride to water according to a certain proportion, and dissolve it for the second time at 80°C to obtain valine salt with a concentration of 5%. (5) valine hydrochloride solution is filtered and desalted through nanofiltration membrane, ion exchange membrane and electrodialysis membrane to obtain valine hydrochloride clear liquid; (6) valine hydrochloride The salt clear solution is first cooled to 60°C for the second time for prelimi...

Embodiment 2

[0009] Example 2: The specific steps for the separation and purification of isoleucine are as follows: (1) filter and sterilize the isoleucine fermentation broth and concentrate it to 5 times, and then spray dry the concentrated fermentation broth to obtain isoleucine crude acid; (2) mix crude isoleucine with hydrochloric acid at a mass ratio of 1:3, and dissolve for the first time at 80°C to obtain crude isoleucine solution; (3) crude isoleucine The solution is cooled to 25°C for the first time and then filtered to obtain isoleucine hydrochloride; (4) Add isoleucine hydrochloride to water in a certain proportion and dissolve for the second time at 80°C to obtain the concentration It is 7% isoleucine hydrochloride solution; (5) the isoleucine hydrochloride solution is filtered and desalted through nanofiltration membrane, ion exchange membrane and electrodialysis membrane to obtain isoleucine hydrochloride clear (6) Lower the temperature of isoleucine hydrochloride clear solut...

Embodiment 3： specific ：(1) to 5，， to ；(2) and 1:3，80℃ no. 1 ， to ；(3) no. 1 to 25℃， to ；(4) 1 proportion ，80℃ no. 2 ， to 4％；(5)、、， to ；(6) no. 2 to 60℃(20min)，60℃(2h)， no. 3 to 50℃ 2 (4h)， no. 4 to 20℃(20min)， to and 。 to and and test 。 test ：89.7％、99.9％。 comparative example 1： specific ：(1) to 5，， to ；(2) and 1:3，80℃ no. 1 ， to ；(3) no. 1 to 25℃， to ；(4) 1 proportion ，80℃ no. 2 ， to 5％；(5)、、， to ；(6) to 50℃50℃(2h、4h)， to and 。 to and and test 。 test ：74.2％、99.6％。 comparative example 2： specific ：(1) to 5，， to ；(2) and 1:3，80℃ no. 1 ， to ；(3) no. 1 to 25℃， to ；(4) 1 proportion ，80℃ no. 2 ， to 5％；(5)、、， to ；(6) no. 2 to 60℃(20min)，60℃(2h)， no. 3 to 50℃(4h)， to and 。 to and and test 。 test ：83.6％、99.6％。 comparative example 3： specific ：(1) to 5，， to ；(2) and 1:3，80℃ no. 1 ， to ；(3) no. 1 to 25℃， to ；(4) 1 proportion ，80℃ no. 2 ， to 5％；(5)、、， to ；(6) no. 2 to 60℃(1h)，60℃(2h)， no. 3 to 50℃ 2 (4h)， no. 4 to 20℃(20min)， to and 。 to and and test 。 test ：84.3％、99.9％。 comparative example 4： specific ：(1) to 5，， to ；(2) and 1:3，80℃ no. 1 ， to ；(3) no. 1 to 25℃， to ；(4) 1 proportion ，80℃ no. 2 ， to 5％；(5)、、， to ；(6)60℃(2h)， no. 2 to 50℃(4h)， no. 3 to 20℃(20min)， to and 。 to and and test 。 test ：80.2％、99.9％。 comparative example 5

[0010]Example 3: The specific steps for the separation and purification of isoleucine are as follows: (1) filter and sterilize the leucine fermentation broth and concentrate it to 5 times, and then spray-dry the concentrated fermentation broth to obtain crude leucine ; (2) Mix the crude leucine and hydrochloric acid according to the mass ratio of 1:3, and dissolve for the first time at 80°C to obtain the crude leucine solution; (3) Cool the crude leucine solution for the first time After reaching 25°C, filter to obtain leucine hydrochloride; (4) add leucine hydrochloride into water according to a certain ratio, and dissolve it for the second time at 80°C to obtain leucine hydrochloride with a concentration of 4%. Hydrochloride solution; (5) leucine hydrochloride solution is filtered and desalted through nanofiltration membrane, ion exchange membrane and electrodialysis membrane to obtain leucine hydrochloride clear liquid; (6) leucine hydrochloride The acid salt clear solution...

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Abstract

The invention embodiment discloses a method for isolating and purifying branched-chain amino acid from fermented liquid, and belongs to the technical field of microbial fermentation. The method disclosed by the invention utilizes four-stage variable temperature crystallization to replace a conventional crystallization technology and comprises the following steps: firstly, carrying out second-timetemperature reduction on branched-chain amino acid hydrochloride clear liquid to 60 to 80 DEG C for preliminary crystallization; secondly, carrying out constant temperature crystal growing under the constant temperature condition of 60 to 65 DEG C; thirdly, carrying out third-time temperature reduction to 50 to 55 DEG C for second-time crystallization; finally, carrying out fourth-time temperaturereduction to 10 to 30 DEG C for final crystallization to obtain a finished product of branched amino acid and hydrochloride thereof. The recovery rate of the finished product of branched amino acid and hydrochloride thereof prepared with the method disclosed by the invention can reach up to 90.1 percent and the purity of the branched amino acid can reach up to 99.9 percent.

Description

technical field [0001] The invention relates to a method for separating and purifying branched-chain amino acids from fermentation broth, and belongs to the technical field of microbial fermentation. Background technique [0002] Branched-chain amino acids refer to the three common amino acids found in proteins, consisting of leucine, valine, and isoleucine. This type of amino acid can promote the release of insulin and the release of growth hormone, which are two special ways to promote human anabolism and accelerate human muscle growth. Therefore, there is a huge market for BCAAs in sports supplements and health products. At present, the production of branched-chain amino acids mainly relies on microbial fermentation, but other amino acids will be produced during microbial fermentation. The molecular weight and chemical isoelectric point of these amino acids, especially neutral amino acids (such as leuc amino acids), are very close to branched-chain amino acids. This cau...

Claims

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Application Information

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IPC IPC(8): C07C227/40C07C229/08
CPCC07C227/40C07C229/08
Inventor 丁杰
Owner 丁杰