CN114805103B - Extraction method of L-alanine - Google Patents

Extraction method of L-alanine Download PDF

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Publication number
CN114805103B
CN114805103B CN202110130338.9A CN202110130338A CN114805103B CN 114805103 B CN114805103 B CN 114805103B CN 202110130338 A CN202110130338 A CN 202110130338A CN 114805103 B CN114805103 B CN 114805103B
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alanine
chromatographic column
mother liquor
chromatographic
purity
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CN114805103A (en
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朱亮
付松
崔怀成
冯学磊
王瑞昌
闻龙
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Anhui Fengyuan Biotechnology Co ltd
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Anhui Fengyuan Biotechnology Co ltd
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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/38Separation; Purification; Stabilisation; Use of additives
    • C07C227/40Separation; Purification
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07BGENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07Optical isomers

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  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

The invention provides an extraction method of L-alanine. The extraction method comprises the following steps: purifying the L-alanine mother liquor by using a circulating chromatographic system to obtain an extract; wherein the purity of the L-alanine contained in the L-alanine mother liquor is 30-80%, preferably 30-50%. The method for extracting the L-alanine can extract the mother liquor of the L-alanine with the purity of less than 50 percent, and can improve the total extraction yield and purity of the L-alanine.

Description

Extraction method of L-alanine
Technical Field
The invention belongs to the technical field of L-alanine production, and relates to an extraction method of L-alanine.
Background
Alanine, although a non-essential amino acid, is the highest amino acid content in human blood, and can provide an amino donor for transaminase, and is often added to transfusion in clinic; in the food industry, L-alanine is popular and appreciated by people as a sweetener and an flavoring agent; in the pharmaceutical industry, L-alanine is an important raw material for producing vitamin B6 and L-aminopropanol, and in addition, in the aspect of high polymer materials, the addition of L-alanine into polylactic acid can effectively improve the performance of the polylactic acid, so that the future market potential of the polylactic acid cannot be estimated.
The production method of L-alanine mainly comprises a protein hydrolysis extraction method, a fermentation method extraction method and an enzyme conversion method. The cost for producing the L-alanine by the fermentation method is low, and accords with the green and healthy consumption concept, so that the method has wide market prospect; after the fermentation process for producing L-alanine is finished, the L-alanine is generally purified by adopting a multiple crystallization method, but the existing extraction process cannot effectively improve the yield of the L-alanine by means of concentration, crystallization and the like, and when the purity of mother liquor is reduced to below 50%, the material has the defects of difficult crystallization, low crystallization rate, powdery crystal particles and the like.
Therefore, it is desirable to provide a novel method by which L-alanine can be extracted using a mother liquor having a low purity, so as to improve the yield and purity of L-alanine.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention aims to provide the method for extracting the L-alanine, which can extract the mother liquor of the L-alanine with the purity of less than 50 percent and can improve the total extraction yield and the purity of the L-alanine.
To achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a method for extracting L-alanine, the method comprising: purifying the L-alanine mother liquor by using a circulating chromatographic system to obtain an extract;
wherein the purity of L-alanine contained in the L-alanine mother liquor is 30 to 80%, preferably 40 to 50%, for example 42%, 45%, 48%, etc.
According to the invention, the components of the L-alanine mother liquor dissolved in the mobile phase have different retention time in the chromatographic system, and the circulating chromatographic system is adopted to purify the L-alanine mother liquor, so that the mother liquor with the L-alanine purity lower than 50% can still be continuously crystallized to obtain the L-alanine, the waste of the L-alanine can be avoided, and the yield and purity of the L-alanine are improved.
The purity of the invention refers to the purity of the L-alanine in the L-alanine mother liquor, wherein the mass ratio of the L-alanine in all materials except the solvent in the L-alanine mother liquor is the purity of the L-alanine in the L-alanine mother liquor.
The circulating chromatographic system comprises more than two chromatographic columns which are sequentially communicated end to end, wherein the last chromatographic column is connected with the first chromatographic column end to end, preferably the last chromatographic column is connected with the first chromatographic column end to end through a circulating pump, the side surface of each chromatographic column is also provided with a feed inlet and a discharge outlet, and preferably the side surface of each chromatographic column is provided with two feed inlets and two discharge outlets.
As a preferable technical scheme of the invention, the circulating chromatographic system comprises 5-7 chromatographic columns which are sequentially communicated end to end, the last chromatographic column is connected with the first chromatographic column end to end through a circulating pump, and two feed inlets and two discharge outlets are also arranged on the side surface of each chromatographic column.
When each component of L-alanine mother liquor is dissolved in a mobile phase and passes through a stationary phase filled in a chromatographic column, the L-alanine mother liquor can be sequentially flowed out of the stationary phase due to different sizes and strengths of the components which are reacted with the stationary phase (adsorption, distribution, ion attraction, exclusion and affinity), the L-alanine mother liquor can be enriched in different chromatographic columns with other substances such as saccharides and inorganic salts in the mother liquor, and an extracting solution enriched in L-alanine and a raffinate enriched in most of the saccharides and inorganic salt components can be obtained through switching different sample outlets.
The invention utilizes a circulating chromatographic system to extract the mother liquor of the L-alanine, so that the purity of the L-alanine in the mother liquor can be improved to 80 percent or even more than 85 percent, and the purity of the L-alanine in the raffinate can be reduced to 30 percent or even less than 15 percent.
The method for purifying the L-alanine mother liquor by using the circulating chromatographic system comprises the following steps: and (3) selecting any chromatographic column for sample injection to perform circulating operation, stopping circulating until the purity of L-alanine in circulating liquid in at least one chromatographic column is more than 80%, eluting the inside of the chromatographic column to obtain extracting liquid, and performing subsequent extraction on the extracting liquid.
As a preferred embodiment of the present invention, the method for eluting the inside of the chromatographic column comprises: and injecting the eluent from an injection port arranged on the side surface of the chromatographic column, and flowing out the extracting solution from an injection port arranged on the side surface of the chromatographic column.
As a preferred technical scheme of the invention, the method for purifying the L-alanine mother liquor by using the circulating chromatographic system comprises the following steps: selecting any chromatographic column for sample injection to perform circulation operation until the purity of L-alanine in the circulating liquid in at least one chromatographic column is more than 80%, and stopping circulation after the purity of L-alanine in the circulating liquid in at least one chromatographic column is less than 15%; eluting the chromatographic column of the circulating liquid with the purity of L-alanine being more than 80% to obtain an extracting solution, and discharging the circulating liquid with the purity of L-alanine being less than 15% to obtain a raffinate.
As a preferred embodiment of the present invention, the elution method includes: introducing an eluent from a sample inlet arranged on the side surface of a chromatographic column with the purity of L-alanine being more than 80%, and flowing out an extracting solution from a sample outlet arranged on the side surface of the chromatographic column;
the invention uses the eluent to push out the circulating liquid in the chromatographic column with the purity of L-alanine above 80%, and uses the eluent to elute the L-alanine adsorbed on the stationary phase.
As a preferred embodiment of the present invention, the method for discharging raffinate includes: introducing the eluent and/or L-alanine mother liquor from a sample inlet arranged on the side surface of the previous chromatographic column of the chromatographic column with the purity of L-alanine below 15%, and discharging the raffinate from a sample outlet arranged on the side surface of the chromatographic column with the purity of L-alanine below 15%.
The invention can eject the circulating liquid in the chromatographic column with the purity of L-alanine below 15% by using the eluent and/or the L-alanine mother liquor, and the eluent and/or the L-alanine mother liquor are preferably injected from the injection port arranged on the side surface of the previous chromatographic column of the chromatographic column with the purity of L-alanine below 15% in the process of discharging the raffinate, so that the aim of ejecting the raffinate can be mainly achieved.
According to the invention, the raffinate is ejected by using the L-alanine mother liquor, so that the purpose of discharging the raffinate can be realized on one hand, and the purpose of sample injection can be realized on the other hand, so that the step of additionally adding sample injection can be omitted, and the next cycle is directly performed after the raffinate is discharged.
As a preferred embodiment of the present invention, the raffinate is discharged twice, and the method comprises: introducing the L-alanine mother liquor from a sample inlet arranged on the side surface of the previous chromatographic column of the chromatographic column with the purity of L-alanine below 15%, and discharging part of raffinate from a sample outlet arranged on the side surface of the chromatographic column with the purity of L-alanine below 15%; the eluent is injected from a sample injection port arranged on the side surface of the chromatographic column which is subjected to elution treatment, and the residual raffinate is discharged from a sample outlet arranged on the side surface of the chromatographic column with the purity of L-alanine below 15 percent.
The circulation operation rate is 0.1-1m/min, such as 0.2, 0.4, 0.5, 0.6, 0.8, etc., and the circulation operation time is preferably 8-10min, such as 8.2min, 8.5min, 8.8min, 9min, 9.2min, 9.5min, 9.8min, etc.
The invention controls the flow rate and the circulation time, so that each component in the mother liquor is just respectively enriched in different chromatographic columns, and the effect of purifying the L-alanine can not be achieved if the flow rate is too slow or too fast.
The extraction method of the invention also comprises the following steps: the dry matter concentration of the L-alanine mother liquor is adjusted to 200-230g/L, the pH value is 6.5-7.0, the temperature is 55-75 ℃, and then the purification is carried out by using a circulating chromatographic system.
The method of the invention further comprises the following steps: and (3) carrying out subsequent extraction on the extracting solution purified by the circulating chromatographic system.
As a preferred embodiment of the present invention, the subsequent extraction step includes decoloring, crystallizing and drying.
The resin filled in the circulating chromatographic system is CSR-2 sodium-based cationic chromatographic resin.
As a preferred embodiment of the present invention, the method for treating a resin comprises: washing with deionized water until the eluate is clear, soaking with 2-2.5mol/L NaOH solution for 20-24h, and washing with distilled water until the eluate is neutral.
Compared with the prior art, the invention has the following beneficial effects:
(1) The extraction method provided by the invention can purify the mother liquor with the purity of L-alanine being lower than 50% to the purity of L-alanine being more than 80%, so that the obtained extract can be continuously concentrated and crystallized;
(2) The extraction method provided by the invention can improve the total yield and purity of the L-alanine, and the total yield can reach more than 90%.
Detailed Description
The technical scheme of the invention is further described by the following specific embodiments. It should be apparent to those skilled in the art that the detailed description is merely provided to aid in understanding the invention and should not be taken as limiting the invention in any way.
Example 1
The embodiment provides an extraction method of L-alanine.
(1) Will be 10m 3 Adding mother liquor (purity is less than or equal to 50%) into 15m 3 Regulating dry matter to 220g/L with pure water (conductance less than or equal to 20 mu s/cm) in a stainless steel storage tank, heating to 65 ℃ with steam, regulating pH to 6.5-7.0 with 30% liquid alkali, sampling and detecting for later use;
(2) Repeatedly soaking and washing resin with deionized water until the eluate is clarified, soaking with 2mol/L NaOH solution for about 24h, washing with distilled water to neutrality, taking 90 tons of resin for use after washing to neutrality, loading into chromatographic columns with phi 3300mm, sequentially connecting the head and the tail of 6 chromatographic columns, respectively marking as No. 1-6 chromatographic columns, and connecting the last chromatographic column with the first chromatographic column end to end through a circulating pump, wherein the side surface of each chromatographic column is also provided with two feed inlets and two discharge outlets;
(3) A centrifugal pump is selected to input the adjusted mother liquor into a chromatographic column for operation, and the primary circulation of the mother liquor is 4.5m 3 Controlling the circulation flow rate to be 0.5m/min, wherein the circulation time is 560 seconds, and after the circulation is stopped, circulating liquid with the purity of L-alanine being more than 80% is positioned on a No. 1 chromatographic column, and circulating liquid with the purity of L-alanine being less than 15% is positioned on No. 4 and No. 5 chromatographic columns;
(4) Feeding 3m from the sample inlet of the No. 4 chromatographic column 3 Mother liquor, discharging (ejecting) raffinate 3m from sample outlet of No. 5 chromatographic column 3 A mother liquor;
(5) Sample inlet of No. 1 chromatographic columnFeeding 6.5m 3 Eluting with pure water from column 1, and discharging 6.5m from the sample outlet 3 Is prepared from the extract of (a);
(6) 3.5m from the sample inlet of chromatographic column 1 3 Is discharged 3.5m from the sample outlet of the No. 5 chromatographic column 3 Is a raffinate from (a) a raffinate from (b);
(7) After the end, the next cycle is started according to the circulating flow rate of 0.5m/min and the circulating time of 560 seconds until the flow rate of 10m 3 Is purified in its entirety.
(8) And (3) decoloring the extracting solution in the step (5) by using active carbon, concentrating, crystallizing, centrifuging and drying to obtain the L-alanine.
Examples 2 to 4
The embodiment provides an extraction method of L-alanine.
Referring to example 1, three different mother liquors were respectively selected for extraction.
Example 5
The embodiment provides an extraction method of L-alanine.
Referring to example 1, a mother liquor having a purity of 32.07% was selected for extraction.
Comparative example 1
The comparative example provides a method for extracting L-alanine.
The mother liquor is separated by concentrating, crystallizing and centrifuging according to the existing mother liquor purification method.
(1) Preparation of 10m 3 Adding mother liquor (purity is less than or equal to 50%) into 15m 3 Sampling and detecting in a stainless steel storage tank for standby;
(2) Transferring the mother liquor to a single-effect evaporator by selecting a centrifugal pump, controlling the vacuum degree of the evaporator to be-0.07-0.075 MPa, and concentrating for 10 hours at 70-72 ℃ for later use;
(3) Separating the concentrated mother liquor by using a 1250-type flat plate centrifuge, separating for 30min by adopting a rotating speed of 900r/min after the feeding of the centrifuge is finished, and cleaning for 5min by using pure water;
(4) Returning the separated crystals to the previous working procedure by using pure water as a melting material, and detecting;
(5) The separated mother liquor is collected and detected.
Performance testing
The extract and raffinate provided in the examples and comparative examples were tested for L-alanine content, and the results are shown in tables 1 and 2:
TABLE 1
TABLE 2
According to the embodiment and the performance test, the extraction method provided by the invention can be used for extracting the L-alanine in the mother liquor with the purity of less than 50%, and the purity and the yield are higher.
As can be seen from the comparison between the example 1 and the comparative example 1, compared with the method in the prior art, the extraction method provided by the invention can successfully obtain L-alanine, has higher yield and avoids the waste of mother liquor with lower purity.
While the invention has been described in detail in the foregoing general description, embodiments and experiments, it will be apparent to those skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.

Claims (1)

1. An extraction method of L-alanine comprises the following steps:
(1) 10m with the purity of 50.24%, 49.63% and 49.11% 3 L-alanine mother liquor was added to 15m 3 The dry matter concentration is adjusted to 220g/L by pure water with the electric conductivity less than or equal to 20 mu s/cm in a stainless steel storage tank, the dry matter concentration is heated to 65 ℃ by steam, the pH value is adjusted to 6.5-7.0 by 30% liquid alkali, and sampling and detection are carried out for standby;
(2) Repeatedly soaking and washing resin with deionized water until the eluate is clarified, soaking for 24 hours with 2mol/L NaOH solution, washing with distilled water to neutrality, taking 90 tons of resin for standby after washing to neutrality, loading into phi 3300mm chromatographic columns, sequentially connecting the 6 chromatographic columns from head to tail, respectively marking as No. 1-6 chromatographic columns with the total length of 8600mm, connecting the last chromatographic column with the first chromatographic column from head to tail through a circulating pump, and arranging two feed inlets and two discharge outlets on the side surface of each chromatographic column;
(3) Selecting centrifugal pump to input the adjusted mother liquor into chromatographic column for operation, and primarily circulating the mother liquor into 4.5m 3 Controlling the circulation flow rate to be 0.5m/min, wherein the circulation time is 560 seconds, and after the circulation is stopped, circulating liquid with the purity of L-alanine being more than 80% is positioned on a No. 1 chromatographic column, and circulating liquid with the purity of L-alanine being less than 15% is positioned on No. 4 and No. 5 chromatographic columns;
(4) Feeding 3m from the sample inlet of the No. 4 chromatographic column 3 Mother liquor, discharging raffinate 3m from sample outlet of No. 5 chromatographic column 3 A mother liquor;
(5) From the sample inlet of the No. 1 chromatographic column, 6.5m 3 Eluting with pure water from column 1, and discharging 6.5m from the sample outlet 3 Is prepared from the extract of (a);
(6) 3.5m from the sample inlet of chromatographic column 1 3 Is discharged 3.5m from the sample outlet of the No. 5 chromatographic column 3 Is a raffinate from (a) a raffinate from (b);
(7) After the end, the next cycle is started according to the circulating flow rate of 0.5m/min and the circulating time of 560 seconds until the flow rate of 10m 3 Purifying the mother liquor of the process;
(8) Decolorizing the extracting solution obtained in the step (5) by using active carbon, concentrating, crystallizing, centrifuging and drying to obtain L-alanine;
wherein: the resin filled in the circulating chromatographic system is CSR-2 sodium-based cationic chromatographic resin.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1950327A (en) * 2004-04-07 2007-04-18 旭化成化学株式会社 Method of purifying amino acid
CN103951573A (en) * 2009-12-29 2014-07-30 罗门哈斯公司 Process for the separation of organic and amino acids from fermentation broths
CN105348122A (en) * 2015-10-30 2016-02-24 安徽丰原发酵技术工程研究有限公司 Method for purifying L-alanine final mother liquor
CN109775911A (en) * 2017-11-13 2019-05-21 秦皇岛华恒生物工程有限公司 A method of l-Alanine mother liquor is handled using Simulation moving bed

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1950327A (en) * 2004-04-07 2007-04-18 旭化成化学株式会社 Method of purifying amino acid
CN103951573A (en) * 2009-12-29 2014-07-30 罗门哈斯公司 Process for the separation of organic and amino acids from fermentation broths
CN105348122A (en) * 2015-10-30 2016-02-24 安徽丰原发酵技术工程研究有限公司 Method for purifying L-alanine final mother liquor
CN109775911A (en) * 2017-11-13 2019-05-21 秦皇岛华恒生物工程有限公司 A method of l-Alanine mother liquor is handled using Simulation moving bed

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
模拟移动床技术分离缬氨酸和丙氨酸;万红贵 等;《食品与发酵工业》;第31卷(第12期);第50-53页 *

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