CN101503366B - Method for extracting and separating L-valine by membrane separation and plant chromatography separation - Google Patents
Method for extracting and separating L-valine by membrane separation and plant chromatography separation Download PDFInfo
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- CN101503366B CN101503366B CN2009100258313A CN200910025831A CN101503366B CN 101503366 B CN101503366 B CN 101503366B CN 2009100258313 A CN2009100258313 A CN 2009100258313A CN 200910025831 A CN200910025831 A CN 200910025831A CN 101503366 B CN101503366 B CN 101503366B
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Abstract
The invention relates to a method for extracting L-valine by combining membrane separation and industrial chromatography separation, belonging to the field of biochemical engineering. The method comprises the steps: (1) thalli is removed by fermentation liquor; (2) inorganic salt and other amino acids are efficiently separated by industrial chromatography; (3) impurity and pigment are removed by the fermentation liquor; (4) the fermentation liquor is concentrated and crystallized, etc. The method has the characteristics of mild reaction condition, simple and convenient operation, less separating steps, good selectivity, clean production and the like, overcomes the defects of lower yield, large sewage discharge capacity and high production intensity in the prior art, and ensures the yield and quality of the L-valine to be obviously improved.
Description
Technical field
The present invention relates to a kind of extraction and separation method of L-Xie Ansuan, relate in particular to a kind of method of utilizing membrane sepn to separate combination technique separation and Extraction L-Xie Ansuan from fermented liquid with industrial chromatography.Belong to biological chemical field.
Background technology
The L-Xie Ansuan is the necessary amino acid of human body as one of branched-chain amino acid, has different physiological roles.Because of its particular structural and function, in human life's metabolism, account for the status of particularly important, mainly in order to the transfusion of preparation aminoacids complex, synthetic polypeptide medicaments and food antioxidant etc., the especially effect in medical research and treatment comes into one's own day by day.It is in the treatment of hemato encephalic barrier, hepatic coma, chronic liver cirrhosis and renal failure; The dietary therapy that inborn errors of metabolism is sick; The surgical wound healing is accelerated in septicemia and postoperative diabetic subject's treatment, is widely used in the nutritional support treatment of tumour patient.Therefore, have application and commercial value widely at medicine and food service industry.
The working method of L-Xie Ansuan has extraction method, chemical synthesis, fermentation method.Extraction method and chemical synthesis are because raw material sources are restricted, and production cost is high, and contaminate environment is difficult to realize suitability for industrialized production.After Japan consonance fermentation company in 1956 was with fermentative Production L-glutamic acid, amino acid whose fermentative prodn development was very fast, and up to the present, most amino acid can be used fermentation method and Production by Enzymes.Production by Microorganism Fermentation L-Isoleucine has advantages such as raw materials cost is low, reaction conditions is gentle, easy realization of large-scale production, is to produce the topmost method of L-Xie Ansuan at present.
At present, ion exchange process is generally adopted in the extraction of L-Xie Ansuan.The general process of this technology is: filtering fermentation liquor is removed thalline, is adopted this technology separation and Extraction L-Xie Ansuan from fermented liquid to have following problem: the one, and adopt filtering method to remove thalline, because thalline is tiny, thalline is removed not thorough usually; The 2nd, L-valine fermentation liquid colour is than higher, and common activated carbon decolorizing is incomplete and consumption is big, influences quality product and yield; The 3rd, impurity component is many in the L-valine fermentation liquid, content is high; Directly use ion exchange method to handle, resin stain is serious, causes that easily resin structure breaks ring and loses regenerative power; Increased running cost; High-load ionic substance can make resin saturated fast simultaneously, increases the resin regeneration number of times, causes problems such as quantity of wastewater effluent is big.
Domestic L-Xie Ansuan fermentative prodn process for extracting is continued to use aforesaid method basically; Indivedual operations are adjusted, but fundamentally do not solve problems such as the fermented liquid Impurity removal is not thoroughly in the L-Xie Ansuan leaching process, ion exchange resin is seriously polluted, L-Xie Ansuan extract yield is low, L-Xie Ansuan product purity is low.
Above-mentioned L-Xie Ansuan extraction process is to set up the 1980s.Along with the progress of biochemical stripping technique, various advanced persons' separating device and technology are played a greater and greater role aborning, and the particularly development and the application of membrane separation technique and industrial chromatography stripping technique greatly taken on a new look various biochemical separating technologies.Membrane separation technique is used for the existing successful precedent of separation and purification of leavened prod, but utilizes membrane sepn to separate combination technique separation and Extraction L-Isoleucine from fermented liquid with industrial chromatography, does not also appear in the newspapers.
Summary of the invention
Deficiency to prior art; The problem that the present invention will solve provides membrane sepn separates combination technique separation and Extraction L-Xie Ansuan from fermented liquid with industrial chromatography method; This method have mild condition, easy and simple to handle, separating step is few, selectivity good; The characteristics of cleaner production environmental protection, the yield that has overcome the prior art existence is not high, sewage flow rate reaches the big shortcoming of production intensity greatly, and the yield of L-Xie Ansuan and quality are significantly improved.
Technical scheme of the present invention: basic step is: at first utilize the millipore filtration of aperture 0.1-0.6 μ m to remove thalline fermented liquid; Obtain fermentation clear liquid through industrial chromatography post separating inorganic salts and part heteroacid; Remove pigment and small molecular weight impurity with nf membrane again; The flaxen L-Xie Ansuan of the clarification that obtains solution makes bullion through steps such as RO reverse osmosis membrane pre-concentration, crystallizations, and bullion obtains highly purified L-Xie Ansuan product after refining.
A kind of method of utilizing membrane sepn to separate combination technique extraction separation L-Xie Ansuan from fermented liquid with industrial chromatography, form by following steps:
(1) fermented liquid heat sterilization body: with fermented liquid through holding back the 0.1-0.6 μ m millipore filtration of relative molecular mass 100,000-1,000,000 D; Thoroughly remove the thalline in the fermented liquid; Obtain L-Xie Ansuan fermentation clear liquid; Working pressure is 0.3-0.8MPa, and service temperature is 10-60 ℃, and the dialysis water yield accounts for the 10%-90% of charging fermentating liquid volume in the volume percentage;
(2) industrial chromatography separates: the L-Xie Ansuan fermentation clear liquid pre-concentration that step (1) is obtained extremely contains thing 3%-6% admittedly; Last industrial chromatography post; Doing the boosting agent with the water of 1-3 times of fermentation clear liquid volume separates; The service temperature 20-50 ℃ of heteroacid of removing in inorganic salt and the fermented liquid obtains highly purified separate fermentation clear liquid;
(3) small molecular protein impurity and pigment are removed in nanofiltration: the L-Xie Ansuan clear liquid after the high purity separation that step (2) is obtained uses holds back nf membrane removal pigment and the small molecular protein impurity of relative molecular mass as 100-3000D; Obtain clarifying colourless L-Xie Ansuan solution; Working pressure is 1.0-3.0MPa; Service temperature is 10-60 ℃, and the dialysis water yield accounts for the 10%-90% of charging clear liquid volume in the volume percentage;
(4) fermented liquid reverse osmosis membrane pre-concentration: step (3) is obtained clarifying flaxen L-Xie Ansuan solution concentrate through reverse osmosis membrane; Admittedly contain thing is the concentrated feed liquid of 3%-7%; Working pressure is 0.5-2.0MPa; Service temperature is 10-60 ℃, and the dialysis water yield accounts for the 10%-90% of feeding liquid volume in the volume percentage;
(5) with ordinary method concentrate, crystallization obtains L-Xie Ansuan bullion.
The service temperature of the said fermented liquid heat sterilization of step (1) body is selected 20-40 ℃ for use.
The described industrial chromatography of step (2) separates, and service temperature is selected 20-40 ℃ for use.
The described nanofiltration of step (3), working pressure is selected 1.0-2.0MPa for use, and service temperature is selected 20-40 ℃ for use, and the dialysis water yield accounts for the 10%-50% of charging clear liquid volume in the volume percentage.
The said reverse osmosis membrane pre-concentration of step (4), working pressure is selected 0.5-1.5MPa for use, and service temperature is selected 20-40 ℃ for use, and the dialysis water yield accounts for the 10%-50% of feeding liquid volume in the volume percentage; Admittedly contain thing is the concentrated feed liquid of 3%-7%.
Beneficial effect of the present invention: the method for separation and Extraction L-Xie Ansuan from fermented liquid that the present invention relates to; First with membrane separation technique introduce fermentation production of L-valine separation and Extraction; And set up one according to the size-grade distribution of impurity in the fermented liquid and overlapped multistage membrane sepn system; The yield that has overcome the prior art existence is not high, quantity of wastewater effluent reaches the high defective of production intensity greatly, and the yield of L-Xie Ansuan and quality are significantly improved, and production cost reduces.
Compare with existing Technology, the meliority of utilizing method of the present invention to have is following:
1, the present invention adopts micro-filtrate membrane filtration to remove the thalline in the fermented liquid, has changed the filtering method of conventional pressure filter.It is thorough to remove thalline through micro-filtrate membrane filtration, crosses the cleaner liquid clear, can also remove high molecular weight protein, and bacteria-eliminating efficacy is much higher than filtration method.
2, the present invention adopts membrane separation technique that ionic impurity and pigment content in the L-valine fermentation liquid are reduced greatly.
3, the industrial chromatography stripping technique has substituted the traditional processing technology of IX, as the boosting agent, has avoided ion exchange regenerant with water, the great amount of wastewater that elution process forms.Brought into play the effect of cleaner production.The high efficiency separation of industrial chromatography simultaneously, other have the amino acid of similar chemical physical property with Xie Ansuan can to remove fermented liquid, and product gas purity obviously improves than the purity that conventional ion exchanges extraction process.
Embodiment
Embodiment 1
L-valine fermentation liquid 4L removes thalline through molecular weight cut-off 200,000 micro-filtrate membrane filtrations, working pressure 0.3MPa, 30 ℃ of temperature, dialysis water yield 1L.Filtrating is thoroughly removed thalline; Industrial chromatography post through 1L separates then, 5 liters of water consumptions, and 30 ℃ of temperature obtain isolating Xie Ansuan clear liquid.Separation clear liquid process molecular weight cut-off is 500 sodium membrane filtration removal small molecular weight impurity and pigment, working pressure 1.2MPa, and 30 ℃ of temperature, dialysis water yield 1.5L passes through RO reverse osmosis membrane pre-concentration then.The result sees the following form.
| T(640nm) | Volume (L) | L-Xie Ansuan (g/L) | Yield (%) | |
| Fermented liquid | 8.3 | 4.0 | 41.1 | 100 |
| Ultra-filtration membrane filtrating | 71.3 | 4.5 | 35.8 | 98.2 |
| The industrial chromatography post | 83.4 | 5.6 | 28.5 | 97.4 |
| The sodium cleaner liquid | 99.5 | 6.0 | 26.5 | 96.1 |
| The RO film | 99.1 | 4.1 | 37.2 | 93.6 |
| Total recovery | 93.6 |
Embodiment 2
L-valine fermentation liquid 300L removes thalline through molecular weight cut-off 800,000 micro-filtrate membrane filtrations, working pressure 0.35MPa, 30 ℃ of temperature, dialysis water yield 90L.Filtrating is thoroughly removed thalline; Industrial chromatography post through 80L separates then, 500 liters of water consumptions, and 30 ℃ of temperature obtain isolating Xie Ansuan clear liquid.Separation clear liquid process molecular weight cut-off is 500 sodium membrane filtration removal small molecular weight impurity and pigment, working pressure 1.2MPa, and 30 ℃ of temperature, dialysis water yield 1.5L passes through RO reverse osmosis membrane pre-concentration then.The result sees the following form.
| T(640nm) | Volume (L) | L-Xie Ansuan (g/L) | Yield (%) | |
| Fermented liquid | 9.1 | 300 | 42.9 | 100 |
| Ultra-filtration membrane filtrating | 75.2 | 334 | 38.1 | 98.9 |
| The industrial chromatography post | 85.3 | 402 | 31.3 | 97.9 |
| The sodium cleaner liquid | 99.2 | 436 | 28.4 | 96.3 |
| The RO film | 98.9 | 300 | 40.6 | 94.6 |
| Total recovery | 94.6 |
Embodiment 3
L-valine fermentation liquid 4.5M
3Remove thalline through molecular weight cut-off 800,000 micro-filtrate membrane filtrations, working pressure 0.4MPa, 30 ℃ of temperature, 1.5 tons of the dialysis water yields.Filtrating is thoroughly removed thalline; Industrial chromatography post through 80L separates then, 7 tons of water consumptions, and 30 ℃ of temperature obtain isolating Xie Ansuan clear liquid.Separation clear liquid process molecular weight cut-off is 1000 sodium membrane filtration removal small molecular weight impurity and pigment, working pressure 1.2MPa, and 30 ℃ of temperature, dialysis water yield 1.5L passes through RO reverse osmosis membrane pre-concentration then.The result sees the following form.
| T(640nm) | Volume (M 3) | L-Xie Ansuan (g/L) | Yield (%) | |
| Fermented liquid | 9.2 | 4.5 | 42.5 | 100 |
| Ultra-filtration membrane filtrating | 75.1 | 5.1 | 36.8 | 98.1 |
| The industrial chromatography post | 83.6 | 6.1 | 30.6 | 97.8 |
| The sodium cleaner liquid | 93.1 | 7.4 | 25.1 | 96.9 |
| The RO film | 99.5 | 5.1 | 35.2 | 94.2 |
| Total recovery | 94.2 |
Embodiment 4
L-valine fermentation liquid 45.8M
3Through removing thalline through molecular weight cut-off 800,000 micro-filtrate membrane filtrations, working pressure 0.35MPa, 30 ℃ of temperature, dialysis water yield 90L.Filtrating is thoroughly removed thalline; Industrial chromatography post through 80L separates then, 700 liters of water consumptions, and 30 ℃ of temperature obtain isolating Xie Ansuan clear liquid.Separation clear liquid process molecular weight cut-off is 500 sodium membrane filtration removal small molecular weight impurity and pigment, working pressure 1.2MPa, and 30 ℃ of temperature, dialysis water yield 1.5L passes through RO reverse osmosis membrane pre-concentration again.The result sees the following form.
| T(640nm) | Volume (M 3) | L-Xie Ansuan (g/L) | Yield (%) | |
| Fermented liquid | 9.5 | 45.8 | 42.6 | 100 |
| Ultra-filtration membrane filtrating | 72.9 | 53.1 | 36.3 | 98.9 |
| The industrial chromatography post | 85.2 | 59.6 | 31.4 | 95.9 |
| The sodium cleaner liquid | 92.8 | 71.3 | 25.8 | 94.3 |
| The RO film | 99.5 | 48.2 | 37.7 | 93.2 |
| Total recovery | 93.2 |
Claims (5)
1. method of utilizing membrane sepn to separate combination technique extraction separation L-Xie Ansuan from fermented liquid with industrial chromatography, form by following steps:
(1) fermented liquid heat sterilization body: with fermented liquid through holding back the 0.1-0.6 μ m millipore filtration of relative molecular mass 100,000-1,000,000 D; Thoroughly remove the thalline in the fermented liquid; Obtain L-Xie Ansuan fermentation clear liquid; Working pressure is 0.3-0.8MPa, and service temperature is 10-60 ℃, and the dialysis water yield accounts for the 10%-90% of charging fermentating liquid volume in the volume percentage;
(2) industrial chromatography separates: the L-Xie Ansuan fermentation clear liquid pre-concentration that step (1) is obtained extremely contains thing 3%-6% admittedly; Last industrial chromatography post; Doing the boosting agent with the water of 1-3 times of fermentation clear liquid volume separates; The service temperature 20-60 ℃ of heteroacid of removing in inorganic salt and the fermented liquid obtains highly purified separate fermentation clear liquid;
(3) small molecular protein impurity and pigment are removed in nanofiltration: the L-Xie Ansuan clear liquid after the high purity separation that step (2) is obtained uses holds back nf membrane removal pigment and the small molecular protein impurity of relative molecular mass as 100-3000D; Obtain clarifying colourless L-Xie Ansuan solution; Working pressure is 1.0-3.0MPa; Service temperature is 10-60 ℃, and the dialysis water yield accounts for the 10%-90% of charging clear liquid volume in the volume percentage;
(4) fermented liquid reverse osmosis membrane pre-concentration: step (3) is obtained clarifying flaxen L-Xie Ansuan solution concentrate through reverse osmosis membrane; Admittedly contain thing is the concentrated feed liquid of 3%-7%; Working pressure is 0.5-2.0MPa; Service temperature is 10-60 ℃, and the dialysis water yield accounts for the 10%-90% of feeding liquid volume in the volume percentage;
(5) with ordinary method concentrate, crystallization obtains L-Xie Ansuan bullion.
2. a kind of method of utilizing membrane sepn to separate combination technique extraction separation L-Xie Ansuan from fermented liquid with industrial chromatography as claimed in claim 1 is characterized in that, the said fermented liquid micro-filtration of step (1) removes the service temperature of thalline and selects 20-40 ℃ for use.
3. a kind of method of utilizing membrane sepn to separate combination technique extraction separation L-Xie Ansuan from fermented liquid with industrial chromatography as claimed in claim 1; It is characterized in that; The described industrial chromatography of step (2) separates, and does the boosting agent with the water of 1-3 times of fermentation clear liquid volume, and service temperature is selected 20-40 ℃ for use.
4. a kind of method of utilizing membrane separation technique extraction separation L-Xie Ansuan from fermented liquid as claimed in claim 1; It is characterized in that; The described nanofiltration of step (3), nf membrane are used and are held back relative molecular mass and be 300-1000D, and working pressure is selected 1.0-2.0MPa for use; Service temperature is selected 20-40 ℃ for use, and the dialysis water yield accounts for the 10%-50% of charging clear liquid volume in the volume percentage.
5. a kind of method of utilizing membrane sepn to separate combination technique extraction separation L-Xie Ansuan from fermented liquid with industrial chromatography as claimed in claim 1; It is characterized in that; The said reverse osmosis membrane pre-concentration of step (4); Service temperature is selected 20-40 ℃ for use, and the dialysis water yield accounts for the 10%-50% of feeding liquid volume in the volume percentage; Admittedly contain thing is the concentrated feed liquid of 3%-7%.
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| CN102161625A (en) * | 2011-03-11 | 2011-08-24 | 无锡晶海氨基酸有限公司 | Method for extracting and separating L-phenylalanine by utilizing combined technology of membrane separation and industrial chromatographic separation |
| CN102432479B (en) * | 2011-12-30 | 2013-11-13 | 通辽梅花生物科技有限公司 | Method for extracting L-valine from L-valine fermentation liquid |
| CN102746174A (en) * | 2012-07-26 | 2012-10-24 | 无锡晶海氨基酸有限公司 | Method for extracting and separating L-valine from fermentation liquor by employing membrane separation and electroosmosis combined technology |
| CN103232353A (en) * | 2013-04-12 | 2013-08-07 | 北京轻发生物技术中心 | Method for separating and extracting L-valine from broth with high efficiency |
| CN104177280A (en) * | 2013-05-23 | 2014-12-03 | 厦门世达膜科技有限公司 | Production process of methionine |
| CN104119241B (en) * | 2014-07-23 | 2017-01-18 | 江苏久吾高科技股份有限公司 | Method for extracting L-leucine |
| CN115141110B (en) * | 2021-03-29 | 2023-12-12 | 安徽华恒生物科技股份有限公司 | Continuous decoloring method of L-valine fermentation liquid |
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| CN1800148A (en) * | 2005-12-12 | 2006-07-12 | 无锡晶海氨基酸有限公司 | Cleaning production process of extracting L-isoleucine from fermented liquor using ion-exchange |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1800148A (en) * | 2005-12-12 | 2006-07-12 | 无锡晶海氨基酸有限公司 | Cleaning production process of extracting L-isoleucine from fermented liquor using ion-exchange |
Non-Patent Citations (1)
| Title |
|---|
| 徐庆阳.金属膜对L-撷氨酸发酵液过滤的研究.《天津科技大学学报》.2006,第21卷(第1期),第4-6页. * |
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Address after: 214100 Donggang Town, Xishan District, Wuxi City, Jiangsu Province Patentee after: WUXI JINGHAI AMINO ACID CO., LTD. Address before: 214199 Donggang Town, Wuxi, Jiangsu Province Patentee before: Wuxi Jinghai Amino Acid Co., Ltd. |