CN101376646A - Novel method for extracting L-tryptophan from fermentation liquor - Google Patents
Novel method for extracting L-tryptophan from fermentation liquor Download PDFInfo
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- CN101376646A CN101376646A CNA2007100593473A CN200710059347A CN101376646A CN 101376646 A CN101376646 A CN 101376646A CN A2007100593473 A CNA2007100593473 A CN A2007100593473A CN 200710059347 A CN200710059347 A CN 200710059347A CN 101376646 A CN101376646 A CN 101376646A
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- tryptophane
- tryptophan
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Abstract
The invention discloses an ion-exchange extraction method for producing L-tryptophan by direct fermentation method, which comprises the following steps: filtering fermenting liquor with a stainless steel tube type membrane separation system, extracting L-tryptophan, and concentrating and crystallizing using the ion-exchange method. The method adopts metal film filter thallus, and improves conventional fermenting liquor treatment method including centrifuging and press filtering in a filter press to reduce the maintenance expense, reduce the labor intensity of workers, remove the thallus thoroughly, and save the operation time. The metal film has the advantages of long service life, resistance to high temperature, acid and alkali, easy cleaning and convenience for device maintenance. The method can greatly improve the extraction efficiency of the L-tryptophan up to acquire total extraction rate of 63.4% and product refined purity up to more than 98%.
Description
[technical field]: the invention belongs to the biochemical engineering field, relate to a kind of method of from fermented liquid, extracting the L-tryptophane.
[background technology]: L-tryptophane formal name used at school tryptophane, chemical name L-2-amino-3-indyl propionic acid, another name L-pancreas Argine Monohydrochloride, L-amino indole propionic acid, molecular formula C11H12O2N2, relative molecular mass 204.21.The L-tryptophane is one of eight kinds of essential amino acids in human body and the animal life activity, is present in the organism with free state or combined.World market L-tryptophane annual requirement is about more than 1,200,000 tons, and the whole world has only more than 1,000 ton throughput at present, and mainly concentrate on a few countries such as Japan, the U.S., only Japan's sales volume in 1989 just reaches 2,500,000,000 yen, thereby the market outlook of L-tryptophane are huge.What have L-tryptophane throughput at present has only minority developed countries such as day, moral, U.S., and China does not carry out suitability for industrialized production, the whole dependence on import in market as yet because of the state of the art restriction.
The production of L-tryptophane mainly relies on proteolysis method and chemical synthesis the earliest, but along with microbial method being produced deepening continuously of L-tryptophane research, microbial method has moved towards practical and has been in dominant position.Microbial method can be divided into direct fermentation, microbe transformation method and enzyme process substantially.Produce L-tryptophane, the industrial capacity that has taken on a certain scale abroad with the microorganism direct fermentation.It is low that this method has a raw materials cost, wide material sources, and the product purity height, advantages such as reaction conditions gentleness have the potentiality of suitability for industrialized production.
In recent years, the L-tryptophane is as indispensable amino acid, use increasingly extensive at aspects such as transfusion, heath food, fodder additivess, extensive application along with methionine(Met), Methionin, the higher always L-tryptophane of price more wishes to have the production and the extracting method of Cheap highly effective, so that advance the application of L-tryptophane.The same with the other biological engineering product, the tryptophane industrial production also usually can be subjected to the restriction of production cost, and in the formation of production cost, the cost of downstream engineerings such as separation and Extraction occupies suitable ratio.Therefore, the extracting method of research L-tryptophane has important significance for theories and practical value.
To really realize the industrialization of fermentation production of L-tryptophan, just must study the extraction process of L-tryptophane in the fermented liquid.Many both at home and abroad employing precipitator method are extracted the L-tryptophane, and existing relevant patent report.The present invention adopts the metal membrane filter fermented liquid, uses L-tryptophane in the ion exchange method separation and Extraction fermented liquid then.
[summary of the invention]: it is low to the objective of the invention is to solve the extract yield that has fermentation production of L-tryptophan now, can't realize the problem that industrialization is produced, and the method for extracting the L-tryptophane from fermented liquid is provided.
A kind of technology of from fermented liquid, extracting the L-tryptophane, comprise: adopt metal membrane filter to remove thalline and albumen, filtrate is extracted L-tryptophane, the pH scope 1～14 of the upper prop feed liquid in the ion exchange process by ion exchange method, service temperature is 0～40 ℃, and flow velocity is 1～6 times of bed volume/h.Ion-exchanging eluent is waited some crystallizations, and service temperature is 0～20 ℃.Mother liquor spent ion exchange resin concentrating return-flow after the crystallization is to waiting in the liquid.Deng some back coarse crystallization deionized water dissolving, add long-pending 50%～95% ethanol of monoploid, refrigeration is spent the night, and carries out recrystallization, gets the crystallization of L-tryptophane after the oven dry.
Metallic membrane adopts stainless steel tubular type film separating system, and its aperture is 50nm～100nm, molecular weight cut-off 2000～50000MW, and pH scope 1～14, service temperature are 0～40 ℃, operation pressure reduction is 0.2～1.0MPa.
Elutriant is 0.1～1mol/L ammoniacal liquor, ammonium chloride solution, ammoniumsulphate soln, Spirit of Mindererus, ethanolic soln.
Ion exchange resin adopts strongly acidic cation-exchange D061, D001-CC, 732, HD-8, JK006, HZ014; Ion exchange column adopts series system.
Before metal membrane filter removes thalline and albumen, fermented liquid carries out pre-treatment, with alkali fermented liquid pH is transferred to 9～12, stirring also is heated to 80 ℃, inject stainless steel tubular type film separating system after adding fermented liquid 0.1%～0.5% diatomite, service temperature is controlled at 50～60 ℃, advances film pressure and membrane pressure difference 0.3～0.8kg/cm
2, filtering velocity is controlled at 200～300ml/min.
Advantage of the present invention and positively effect: the present invention utilizes metal membrane filter method pre-treatment fermented liquid, and extracts the L-tryptophane with ion exchange method, can improve the yield of L-tryptophane greatly, shortens the operational cycle.Product purity can reach more than 98%, extraction yield 63.4%.
The present invention adopts the metal membrane filter thalline, and methods such as centrifugal, the pressure filter press filtration that has changed that conventional fermentation liquor treatment adopts have reduced maintenance cost, have reduced working strength of workers, and it is thorough to remove thalline, saves the operating time; And the metallic membrane long service life, is easy to clean and convenience for device maintenance high temperature resistant, acid and alkali-resistance.
Utilize the metal membrane filter pre-treatment can fast and effeciently remove thalline, foreign protein and pigment etc. in the fermented liquid, help the ion-exchange absorption of L-tryptophane.Ion exchange method can further play decolorization in addition, removes the pigment of filtrate, thereby the operational cycle has been shortened in the decolouring operation of omission employing gac etc.
[extraction process flow process]:
See Figure of description.
[embodiment]:
Embodiment 1: membrane filter method removes albumen and pigment
With sulfuric acid L-tryptophane fermented liquid pH is transferred to 3, stir and 80 ℃ of heating in water bath 10min, add 0.1% diatomite, pour fermented liquid into the metal membrane filter device, 50 ℃～60 ℃ of controlled temperature advance film pressure and membrane pressure difference 0.7kg/cm
2, when turnover mould difference obviously reduces, mend appropriate amount of deionized water, filtering velocity is controlled at 200～300ml/min.Albumen and pigment removal are respectively 94.5% and 50.0%.L-tryptophane rate of loss is 3.2%.
Embodiment 2: the screening of ion exchange resin
For the L-tryptophane in the ion exchange method extraction fermented liquid, exchange capacity is a most important index of weighing the ion exchange resin quality.Experiment has selected for use 6 kinds of storng-acid cation exchange resin D061, D001-CC, 732, HD-8, JK006, HZ014 to carry out the static state experiment, measures the Static Adsorption amount Q of various resins to the L-tryptophane, and the result is as shown in table 1.
Table 1 different model resin is to the Static Adsorption amount of L-tryptophane
As shown in Table 1, resin 732, HD-8, JK006, four kinds of resin absorption amounts of HZ014 are bigger.
Embodiment 3: ion exchange method is extracted the L-tryptophane
The JK006 resin that is processed into the H type is packed in the ion exchange column of diameter 2.5cm, the high 16cm of dress post, service temperature is a room temperature, filtrate or etc. the mother liquor upper prop speed after the some crystallization be 1BV/h, upper column quantity is the twice bed volume.Behind the upper prop, with 1 times of volume water washing resin, flow velocity is 1BV/h, washings is collected prepared upper prop once more.With 1mol/L ammoniacal liquor 1BV/h wash-out, collect the parting liquid of 2～7 times of resin volumes.The ion exchange method extract yield is 90.7%.
The crystallization of embodiment 4:L-tryptophane etc. point
Ion-exchanging eluent elder generation concentrating under reduced pressure is evaporated portion water, obtain certain density tryptophane solution, again supersaturated solution is cooled off, in solution, add 30% ethanolic soln, slowly regulate pH to 6.5 with the vitriol oil, 4 ℃ of temperature are added 0.1% crystal seed, then pH is transferred to 5.88,4 ℃ and wait some crystallization 24h.Deng the some crystallization yield is 91.8%.
Embodiment 5:L-tryptophane is refining
With a certain amount of deionized water dissolving coarse crystallization, add isopyknic 95% ethanol again, refrigeration is spent the night, and filters crystal, gets the crystallization of L-tryptophane after the oven dry, and purity can reach more than 98%, and refining yield is 93.6%.
Claims (5)
1, a kind of technology of from fermented liquid, extracting the L-tryptophane, comprise: adopt metal membrane filter to remove thalline and albumen, filtrate is extracted the L-tryptophane by base exchange method, the pH scope 1～14 of the upper prop feed liquid in the ion exchange process, service temperature is 0～40 ℃, and flow velocity is 1～6 times of bed volume/h.Ion-exchanging eluent is waited some crystallizations, and service temperature is 0～20 ℃.Mother liquor spent ion exchange resin concentrating return-flow after the crystallization is to waiting in the liquid.Deng some back coarse crystallization deionized water dissolving, add long-pending 50%～95% ethanol of monoploid, refrigeration is spent the night, and carries out recrystallization, gets the crystallization of L-tryptophane after the oven dry.
2, production technique according to claim 1 is characterized in that metallic membrane adopts stainless steel tubular type film separating system, and its aperture is 50nm～100nm, molecular weight cut-off 2000～50000MW, pH scope 1～14, service temperature are 0～40 ℃, and operation pressure reduction is 0.2～1.0MPa.
3, want 1 described production technique according to right, it is characterized in that elutriant is 0.1～1mol/L ammoniacal liquor, ammonium chloride solution, ammoniumsulphate soln, Spirit of Mindererus, ethanolic soln.
4, production technique according to claim 1, the ion exchange resin that it is characterized in that adopts strongly acidic cation-exchange D061, D001-CC, 732, HD-8, JK006, HZ014; Ion exchange column adopts series system.
5, production technique according to claim 1, before it is characterized in that filtering, fermented liquid carries out pre-treatment, pH transfers to 1～14 with fermented liquid, stirring also is heated to 80 ℃, inject stainless steel tubular type film separating system after adding fermented liquid 0.1%～0.5% diatomite, service temperature is controlled at 50～60 ℃, advances film pressure and membrane pressure difference 0.3～0.8kg/cm
2, filtering velocity is controlled at 200～300ml/min.
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| CNA2007100593473A CN101376646A (en) | 2007-08-29 | 2007-08-29 | Novel method for extracting L-tryptophan from fermentation liquor |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101717360B (en) * | 2009-12-24 | 2011-09-07 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting L-tryptophan from fermentation liquid |
| CN101709049B (en) * | 2009-11-17 | 2011-09-14 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting L-tryptophan |
| CN101709048B (en) * | 2009-11-11 | 2011-10-26 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting L-tryptophan |
| CN103315999A (en) * | 2013-07-05 | 2013-09-25 | 四川省惠达药业有限公司 | Compound amino acid composition and preparation method thereof |
| CN103709091A (en) * | 2013-11-29 | 2014-04-09 | 大成生化科技(松原)有限公司 | Method for extracting L-tryptophan |
| CN104193668A (en) * | 2014-09-10 | 2014-12-10 | 新疆天富阳光生物科技有限公司 | Method for extracting L-tryptophan from fermentation liquor through macroporous resin |
| CN104447493A (en) * | 2014-11-20 | 2015-03-25 | 河南巨龙生物工程股份有限公司 | Method for extracting L-tryptophan high-efficiency membrane |
| CN104478786A (en) * | 2014-11-20 | 2015-04-01 | 河南巨龙生物工程股份有限公司 | Extracting and energy-saving system for L-tryptophan membrane |
| WO2015104464A1 (en) | 2014-01-07 | 2015-07-16 | Novasep Process | Process for purifying aromatic amino acids |
| CN105566136A (en) * | 2016-01-19 | 2016-05-11 | 天津科技大学 | Method for separating and extracting 4-hydroxyisoleucine from fermentation liquor |
| CN105693592A (en) * | 2016-03-03 | 2016-06-22 | 天津科技大学 | Process method for efficiently extracting L-tryptophan from fermentation liquor through thallus carrying crystallization |
| CN109517858A (en) * | 2018-12-17 | 2019-03-26 | 新疆阜丰生物科技有限公司 | A method of production and extraction L-Trp |
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2007
- 2007-08-29 CN CNA2007100593473A patent/CN101376646A/en active Pending
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101709048B (en) * | 2009-11-11 | 2011-10-26 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting L-tryptophan |
| CN101709049B (en) * | 2009-11-17 | 2011-09-14 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting L-tryptophan |
| CN101717360B (en) * | 2009-12-24 | 2011-09-07 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting L-tryptophan from fermentation liquid |
| CN103315999B (en) * | 2013-07-05 | 2015-04-01 | 四川省惠达药业有限公司 | Compound amino acid composition and preparation method thereof |
| CN103315999A (en) * | 2013-07-05 | 2013-09-25 | 四川省惠达药业有限公司 | Compound amino acid composition and preparation method thereof |
| CN103709091A (en) * | 2013-11-29 | 2014-04-09 | 大成生化科技(松原)有限公司 | Method for extracting L-tryptophan |
| CN103709091B (en) * | 2013-11-29 | 2016-04-06 | 大成生化科技(松原)有限公司 | A kind of method extracting L-Trp |
| WO2015104464A1 (en) | 2014-01-07 | 2015-07-16 | Novasep Process | Process for purifying aromatic amino acids |
| CN104193668A (en) * | 2014-09-10 | 2014-12-10 | 新疆天富阳光生物科技有限公司 | Method for extracting L-tryptophan from fermentation liquor through macroporous resin |
| CN104193668B (en) * | 2014-09-10 | 2016-02-03 | 新疆天富阳光生物科技有限公司 | Macroporous resin extracts the method for L-Trp in fermented liquid |
| CN104447493A (en) * | 2014-11-20 | 2015-03-25 | 河南巨龙生物工程股份有限公司 | Method for extracting L-tryptophan high-efficiency membrane |
| CN104478786A (en) * | 2014-11-20 | 2015-04-01 | 河南巨龙生物工程股份有限公司 | Extracting and energy-saving system for L-tryptophan membrane |
| CN105566136A (en) * | 2016-01-19 | 2016-05-11 | 天津科技大学 | Method for separating and extracting 4-hydroxyisoleucine from fermentation liquor |
| CN105693592A (en) * | 2016-03-03 | 2016-06-22 | 天津科技大学 | Process method for efficiently extracting L-tryptophan from fermentation liquor through thallus carrying crystallization |
| CN105693592B (en) * | 2016-03-03 | 2019-03-19 | 天津科技大学 | A kind of carry disease germs from fermentation liquid crystallizes the process of high efficiency extraction L-Trp |
| CN109517858A (en) * | 2018-12-17 | 2019-03-26 | 新疆阜丰生物科技有限公司 | A method of production and extraction L-Trp |
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