CN104119241A - Method for extracting L-leucine - Google Patents
Method for extracting L-leucine Download PDFInfo
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- CN104119241A CN104119241A CN201410353272.XA CN201410353272A CN104119241A CN 104119241 A CN104119241 A CN 104119241A CN 201410353272 A CN201410353272 A CN 201410353272A CN 104119241 A CN104119241 A CN 104119241A
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Abstract
The invention provides a method for extracting L-leucine. The method comprises the following steps of heating a L-leucine fermentation broth, sterilizing and filtering by virtue of a ceramic ultrafiltration membrane to obtain ceramic membrane ultrafiltration supernatant which is mainly divided into two parts, namely, a front part as a first filtrate and a rear part as a second filtrate obtained by filtering the fermentation broth with the membrane to obtain a concentrated broth and eluting the concentrated broth; adjusting the pH value of the second filtrate with sulfuric acid to be 2-3, and then adsorbing with iron exchange resin; after the resin is adsorbed to be saturated, desorbing with ammonia water of which the pH value is about 10; commonly decolorizing the desorption solution and ceramic ultrafiltration membrane filtrate A with an organic nanofiltration membrane; evaporating and condensing the decolorized solution; cooling, crystallizing and drying to obtain a crude L-leucine product.
Description
Technical field
The present invention relates to a kind of extracting method of L-Leu, relate in particular to adopt membrane filtration combine with traditional technology extraction L-Leu method.
Background technology
L-Leu, formal name used at school is 2-amino-4-methylvaleric acid, molecular formula is (CH
3)
2cHCH
2cH (NH
2) COOH, relative molecular weight 131.18.A kind of nonpolar alpha amino acid of aliphatics side chain that contains 6 carbon atoms.L-Leu is one of common 20 seed amino acids of constitutive protein matter, is mammiferous indispensable amino acid and raw ketone glycogenic amino acid.It and Isoleucine isomers each other.In trophology, leucine is the amino acid the most often occurring in protein, and plays important effect for the nitrogen equilibrium in normal development and grownup's health of baby and childhood.By inference, leucine may maintain on muscle and play an important role in the method for the biochemistry with balance protein and decomposition.
At present, L-Leu production technique falls behind, and product yield is low, purity is low.In prior art, have a kind of technique be will L-Leu fermented liquid ceramic membrane filter liquid mix after through ion-exchange resins absorption, after desorb by stripping liquid directly through row evaporation concentration, after concentrated solution is carried out to activated carbon decolorizing twice, destainer carries out decrease temperature crystalline.In this production technique, the ceramic super-filtering film filtered liquid of fermented liquid is all adsorbed through ion-exchange resins, ion-exchange resins working load is large, and acid, alkali, the water yield that the cleaning of resin column and regeneration are used are large.To after handing over stripping liquid concentrated, need to carry out twice activated carbon decolorizing, activated carbon dosage is large and product yield is lower.Resin column Adsorption and desorption and activated carbon decolorizing operation cause L-Leu yield lower, after crystallization in crude product foreign matter content high.In patent application 201210260546.1, introduce the method for utilizing membrane sepn to separate L-Leu with the extraction of desalination from fermented liquid of electrodialysis combination technique, in the method by L-Leu fermented liquid membrane filtration liquid directly through electrodialysis desalination, because still containing the impurity such as a large amount of albumen in L-Leu membrane filtration liquid, cause electrodialytic desalting speed low, and electrodialysis process to membrane filtration liquid without decolorizing effect.
Summary of the invention
The object of the invention is to overcome the problems such as complex process in prior art, energy consumption is high, L-Leu yield is low, provide a kind of technique simple, easy and simple to handle, can obviously reduce production costs and can improve the quality of products and the L-Leu extracting method of yield.
Technical scheme:
The extracting method of L-Leu, comprises the steps:, by L-Leu fermented liquid ultrafiltration membrance filter, to obtain the first filtrate; Then the first filtrate is sent into separation circuit; Described separation circuit refers to and filters by first nanofiltration membrane, then nanofiltration permeate is obtained to L-Leu by after concentrated, crystallization.
According to an embodiment of the invention, after ultrafiltration membrance filter, water carries out dialysis to the filter cake of ultra-filtration membrane, obtains the second filtrate, then the second filtrate is sent into described separation circuit.
According to an embodiment of the invention, the second filtrate being need to be after ion exchange resin carries out Adsorption and desorption processing, then sends into described separation circuit.
According to an embodiment of the invention, the molecular weight cut-off of described nanofiltration membrane is 500～1000Da.
Beneficial effect:
In the present invention, ceramic super-filtering film filtered liquid is filtered to decolouring through organic nanofiltration membrane, obviously reduced ion-exchange resins consumption, reduced acid, alkali, water consumption, production technique is simpler, and product yield obviously improves, and amount of investment still less.In addition, by the filter cake of ultra-filtration membrane being carried out to dialysis, ion-exchange resins purifying, can improve productive rate, alleviate the working load of nanofiltration membrane.
Embodiment
Below by embodiment, the present invention is described in further detail.But it will be understood to those of skill in the art that the following example is only for the present invention is described, and should not be considered as limiting scope of the present invention.Unreceipted concrete technology or condition person in embodiment, for example, according to the described technology of the document in this area or condition (" mineral membrane isolation technique and the application " of showing with reference to Xu Nan equality, Chemical Industry Press, 2003) or carry out according to product description.The unreceipted person of production firm of agents useful for same or instrument, being can be by the conventional products of commercial acquisition.
Approximate language used herein can be used for modifying any quantity statement in whole specification sheets and claims, and it can permit changing under the condition that does not cause its relevant basic function to change.The value of therefore, being modified by the term such as " approximately " is not limited to specified exact value.In at least some cases, approximate language can be corresponding with the precision of the instrument for measuring this value.Unless separately point out in context or statement, otherwise range limit can combine and/or exchange, and this scope is confirmed as and comprised included all subranges herein.Except in operation embodiment or indicating in elsewhere, the numeral of the amount of all expression compositions that use in specification sheets and claims, reaction conditions etc. or expression all should be understood to be subject to the modification of word " approximately " in all cases.
First the present invention first filters with ultra-filtration membrane L-Leu fermented liquid, remove impurity wherein, albumen, colloid, thalline etc., carry out purification process with the filtrate that nanofiltration membrane is carried out ultrafiltration again, nanofiltration membrane can be got rid of ultrafiltration and see through some pigments, impurity and a part of inorganic salt in liquid, L-Leu is seen through, the effect that product separation is purified can be played, next, L-Leu can be obtained by step concentrated to nanofiltration membrane, recrystallize.
For ultra-filtration membrane of the present invention, as long as can realize such the object of the invention of composition such as removing water-soluble polymer in fermented liquid, thalline, particle, colloid, be not particularly limited, can enumerate: the organic materialss such as Mierocrystalline cellulose, cellulose ester, polysulfones, polyethersulfone, polyvinyl chloride, propenyl chloride, polyolefine, polyvinyl alcohol, polymethylmethacrylate, poly(vinylidene fluoride), tetrafluoroethylene, or the inorganic materials such as the metal such as stainless steel or pottery.Preferably ceramic material.
For the step of ultrafiltration membrance filter, the selection in the aperture of ultra-filtration membrane directly has influence on the effect of filtering fermentation liquor degerming, aperture is excessive, can make filtering accuracy step-down, impurity permeation ceramic membrane in fermented liquid, increase the processing load of decolouring film (nanofiltration membrane), also affect the purity of subsequent product; Aperture is too small, can make filtration resistance excessive, and membrane flux is too low, causes that ceramic membrane filter facility investment is excessive, energy consumption increases, and in addition, also can hold back part effective constituent, causes ultimate yield to reduce.In the present invention, ceramic super-filtering film pore diameter range is 0.02～0.05 μ m, and preferably, ceramic membrane aperture is 0.05 μ m.Working pressure also directly has influence on the steady running of filtration procedure, and working pressure is too high, can cause film to pollute aggravation, causes membrane flux sharp-decay; Working pressure is too low, can make the motivating force of filtration procedure too little, causes membrane flux too low.In the present invention, ceramic membrane working pressure is 0.1～0.3MPa, and preferably, ceramic membrane working pressure is 0.2MPa.If the crossflow velocity in ultra-filtration process is excessive, can cause face cannot form good cake layer, these cake layers can play the effect of holding back further impurity in fermented liquid, can cause fine impurities to see through ultra-filtration membrane; If crossflow velocity is too small, can cause the flux of ultrafiltration too small, cycles of concentration cannot be improved further, cause ultimate yield step-down.In technical scheme of the present invention, the crossflow velocity of ceramic membrane filter is 2～5m/s, and preferably, the crossflow velocity of ceramic membrane filter is 4m/s.In ceramic membrane is concentrated, cycles of concentration is more excellent is 5～20 times, optimum 10 times.
Nanofiltration membrane is the film that is defined as " prevention is less than the particle of 2nm and the macromolecular pressure drive membrane of dissolving " herein.Be applicable to preferably such film of effective nanofiltration membrane of the present invention: on this film surface, have electric charge, thereby the combination that is separated (particle size separation) and had benefited from the electrostatic separation of the lip-deep electric charge of this film by pore shows the separation efficiency of raising.Therefore, essential adopt such nanofiltration membrane, this nanofiltration membrane can be in carrying out separation from other ions with different charge characteristics by electric charge as separate targets product, remove high score subclass material by separation.As the material of the nanofiltration membrane using in the present invention, can use the macromolecular materials such as rhodia based polymer, polymeric amide, SPSF, polyethersulfone, polysulfones, polyacrylonitrile, polyester, polyimide and vinyl polymer, in concrete application system involved in the present invention, optimum is to adopt polyethersulfone (PES) or polysulfones (PS), retaining molecular weight is 500～1000Da, more excellent is 800Da, membrane filtration pressure is 0.5～1.5MPa, more excellent is 1.0MPa, and filtration temperature is 30～45 DEG C; While adopting such nanofiltration, can make to greatest extent L-Leu see through, and hold back inorganic salt and other impurity.The described film that is only made up of a kind of material of being not limited to can be the film that comprises multiple described material.About membrane structure, described film can be asymmetric membrane, and it at least has tight zone in one side film, and has from tight zone to film inside or another side aperture becomes large micropore gradually; Or composite membrane, it has the very thin functional layer being formed by other material on the tight zone of asymmetric membrane.In the present invention, adopt nanofiltration membrane to decolour and desalting treatment to ultra-filtration membrane penetrating fluid, be obviously better than traditional effect that utilizes activated carbon decolorizing; Because destainer does not need again through row activated carbon decolorizing, avoided that the activated carbon dosage that activated carbon decolorizing brings is large, charcoal absorption product causes that product yield is lower, very easily leak carbon in activated carbon decolorizing process causes product purity on the low side and use gac cost high and can be to problems such as environments, and decolouring and desalination are placed in same step and are carried out, there is the simple advantage of step.Nanofiltration membrane destainer transparence can reach more than 70%, L-Leu content 20～30mg/L.
Obtaining nanofiltration through after liquid, when being concentrated, it more preferably adopts evaporation concentration, until carry out decrease temperature crystalline after having mass crystallization to occur, Tc is 15～20 DEG C; Due to L-Leu, at high temperature solubleness is higher, adopts evaporation concentration, compared with other concentration method, can obviously improve the cycles of concentration of feed liquid, and follow-up crystallisation process need be only also that cocoa significantly improves cycles of concentration by decrease temperature crystalline, raises the efficiency.This technique can be increased to product yield more than 95% by 80% left and right, has also reduced production cost simultaneously.In the present invention, the yield of L-Leu is to calculate like this: yield=(weight of L-Leu in L-Leu product weight/fermented liquid) × 100%.
Due to after ultrafiltration membrance filter, in concentrated solution or filter cake, still have a certain amount of L-Leu, in order to improve yield, according to an improved embodiment of the present invention, dialysis adds water in concentrated solution, obtain dialyzate (being called the second filtrate in the present invention), can more the active substance wash-out in filter cake out, can more improve the yield of L-Leu, preferably under condition, amount of water is 5～15 times of concentrated liquid measure, and more excellent is 10 times.In preferred situation, the pH regulator of the water adding during by dialysis is 8～9, can make further the L-Leu in filter cake separate out, and yield is improved.
In dialyzate, the content of L-Leu is lower, while directly entering nanofiltration membrane filtration, can cause working load larger, according to an improved embodiment of the present invention, need to adsorb and wash-out dialyzate spent ion exchange resin, the effect that can produce on the one hand concentrated feed liquid, another aspect is can be to dialyzate purifying further.Can adopt dilute sulphuric acid to regulate dialyzate pH to 2～3, adsorb through ion exchange resin column again, resin column continuously feeding is adsorbable saturated, adsorb the saturated rear ammoniacal liquor with pH10 left and right L-Leu is carried out to desorb, thereby improve the leucine content in filtrate, in ion exchange resin stripping liquid, effective constituent can improve 2～3 times, reach 25～30mg/L, further dwindle material liquid volume, reduce the foreign matter content (also can remove a part of impurity in sorption and desorption process) in feed liquid, thereby alleviate the load of follow-up nanofiltration membrane decolouring.Ion exchange resin can be itself known ion exchange resin, for example Zeo-karb, anionite-exchange resin, and the functional group that these ion exchange resin possess is sulfonic group, carboxylic acid group and quaternary ammonium group etc. respectively.The thickness of ion exchange resin can be 10 μ m～100 μ m, and preferably the hole dimension of 10 μ m～30 μ m. ion exchange resin can be 1 μ m～10 μ m, is preferably 4 μ m～7 μ m.For example ion exchange resin or preferably configured with the thickness of 0.1 μ m～100 μ m by chippy ion exchange resin.The pulverizing of ion exchange resin for example can be by utilizing mortar to pulverize to carry out.In addition, the size of chippy ion exchange resin can be 0.01 μ m～10 μ m.The type of ion exchange resin is preferably storng-acid cation exchange resin, for example 732, WA-2, HD-8, JK008, HZ014, and more preferably 732.After the stripping liquid of ion exchange resin can mix with the filtrate of ultra-filtration membrane, filter by nanofiltration membrane, also can send into respectively nanofiltration membrane and filter.Most of ceramic super-filtering film filtered liquid is directly through organic nanofiltration membrane decolouring, and ion-exchange resins post operation is only processed a small amount of dialyzate, has obviously reduced ion-exchange resins consumption, has reduced acid, alkali, water consumption.Production technique is simpler, and product yield obviously improves, and amount of investment still less.
Embodiment 1
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 20nm, filter pressure 0.3MPa, crossflow velocity 2m/s, after concentrating 7 times, fermented liquid obtains ultrafiltrated permeation liquid, penetrating fluid is through organic nanofiltration membrane decolouring desalination, the material of nanofiltration membrane is polyethersulfone, retaining molecular weight is 500Da, pressure 0.6MPa, 40 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring and product in dope is carried out to wash-out obtains organic membrane destainer 170kg altogether.L-Leu content 26mg/L in destainer, transparence 72.3%.Destainer is cooled to concentrated solution to 18 DEG C after evaporation concentration certain multiple and carries out decrease temperature crystalline, after crystal drying, obtain L-Leu crude product, L-Leu purity 82.3%, yield 75.9%.
Embodiment 2
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 20nm, filter pressure 0.3MPa, crossflow velocity 2m/s, after concentrating 7 times, fermented liquid obtains ultrafiltrated permeation liquid, add wash water to stream in ultrafiltration dope again L-Leu in dope and filter cake is carried out to wash-out, amount of water is 13 times of concentrated solution volume, obtain dialyzate, after ultrafiltrated permeation liquid and dialyzate are mixed through organic nanofiltration membrane decolouring desalination, the material of nanofiltration membrane is polyethersulfone, retaining molecular weight is 500Da, pressure 0.6MPa, 40 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring and product in dope is carried out to wash-out obtains organic membrane destainer 190kg altogether.L-Leu content 30mg/L in destainer, transparence 74.8% is cooled to concentrated solution 18 DEG C by destainer and carries out decrease temperature crystalline after evaporation concentration certain multiple, obtains L-Leu crude product, L-Leu purity 83.8%, yield 80.2% after crystal drying.Can find out, after carrying out the dialysis operation of dope, can make yield improve.
Embodiment 3
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 20nm, filter pressure 0.3MPa, crossflow velocity 2m/s, after concentrating 7 times, fermented liquid obtains ultrafiltrated permeation liquid, add wash water to stream in ultrafiltration dope again L-Leu in dope and filter cake is carried out to wash-out, amount of water is 13 times of concentrated solution volume, obtain dialyzate, dialyzate is regulated to pH to 3 with dilute sulphuric acid, by adsorbing from storng-acid cation exchange resin WA-2 post, adsorb the saturated rear ammoniacal liquor with pH10 the L-Leu on resin column is carried out to desorb, obtain stripping liquid 18kg altogether.After ultrafiltrated permeation liquid and stripping liquid are mixed through organic nanofiltration membrane decolouring desalination, the material of nanofiltration membrane is polyethersulfone, and retaining molecular weight is 500Da, pressure 0.6MPa, 40 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring and product in dope is carried out to wash-out obtains organic membrane destainer 170kg altogether.L-Leu content 40mg/L in destainer, transparence 80.3% is cooled to concentrated solution 18 DEG C by destainer and carries out decrease temperature crystalline after evaporation concentration certain multiple, obtains L-Leu crude product, L-Leu purity 88.7%, yield 89.5% after crystal drying.Can find out, after dialyzate is carried out to resin absorption, wash-out, can improve significantly the transparence of product liquid, also can improve yield and purity.
Embodiment 4
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 50nm, filter pressure 0.1MPa, crossflow velocity 4m/s, after concentrating 7 times, fermented liquid obtains ultrafiltrated permeation liquid, add wash water to stream in ultrafiltration dope again L-Leu in dope and filter cake is carried out to wash-out, amount of water is 8 times of concentrated solution volume, obtain dialyzate, dialyzate is regulated to pH to 2 with dilute sulphuric acid, by adsorbing from storng-acid cation exchange resin 732 posts, adsorb the saturated rear ammoniacal liquor with pH10 the L-Leu on resin column is carried out to desorb, obtain stripping liquid 10kg altogether.After ultrafiltrated permeation liquid and stripping liquid are mixed through organic nanofiltration membrane decolouring desalination, adopt respectively the nanofiltration membrane of unlike material, retaining molecular weight is 1000Da, pressure 1.2MPa, 30 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring carries out wash-out to product in dope and obtains organic membrane destainer 150kg altogether, destainer is cooled to concentrated solution to 18 DEG C after evaporation concentration certain multiple and carries out decrease temperature crystalline, after crystal drying L-Leu crude product.
Product index under the test conditions of different nanofiltration membrane is as following table:
| Nanofiltration membrane material | SPSF | Polymeric amide | Polyethersulfone | Polysulfones | Polyimide | Polyacrylonitrile |
| Destainer transparence % | 70.2 | 71.5 | 81.1 | 81.3 | 72.5 | 71.3 |
| L-Leu purity % | 84.2 | 79.4 | 88.5 | 86.5 | 84.7 | 83.6 |
| L-Leu yield % | 75.4 | 72.4 | 91.5 | 91.2 | 72.4 | 83.7 |
As can be seen from the table, while adopting poly (ether sulfone) film and polysulfone membrane, the purity of the product of gained and yield are all high compared with other nanofiltration membrane.
Embodiment 5
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 50nm, filter pressure 0.1MPa, crossflow velocity 4m/s, after concentrating 7 times, fermented liquid obtains ultrafiltrated permeation liquid, add wash water to stream in ultrafiltration dope again L-Leu in dope and filter cake is carried out to wash-out, amount of water is 8 times of concentrated solution volume, obtain dialyzate, dialyzate is regulated to pH to 2 with dilute sulphuric acid, by adsorbing from storng-acid cation exchange resin 732 posts, adsorb the saturated rear ammoniacal liquor with pH10 the L-Leu on resin column is carried out to desorb, obtain stripping liquid 10kg altogether.After ultrafiltrated permeation liquid and stripping liquid are mixed through organic nanofiltration membrane decolouring desalination, adopt polyether sulfone filtering film, retaining molecular weight adopts scale not of the same race, pressure 1.2MPa, 30 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring carries out wash-out to product in dope and obtains organic membrane destainer 150kg altogether, destainer is cooled to concentrated solution to 18 DEG C after evaporation concentration certain multiple and carries out decrease temperature crystalline, after crystal drying L-Leu crude product.
Product index under the test conditions of different nanofiltration membrane is as following table:
| Nanofiltration membrane molecular weight cut-off | 300 | 400 | 500 | 700 | 800 | 1000 |
| Destainer transparence % | 85.4 | 85.9 | 86.1 | 86.2 | 86.1 | 83.8 |
| L-Leu purity % | 92.5 | 91.9 | 92.5 | 90.2 | 92.6 | 89.5 |
| L-Leu yield % | 84.7 | 85.7 | 87.9 | 93.1 | 95.1 | 95.1 |
As can be seen from the table, while adopting 500～1000 molecular weight cut-off, the purity of the product of gained and yield are all high compared with other nanofiltration membrane.
Embodiment 6
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 50nm, filter pressure 0.2MPa, crossflow velocity 4m/s, after concentrating 10 times, fermented liquid obtains ultrafiltrated permeation liquid, add wash water to stream in ultrafiltration dope again L-Leu in dope and filter cake is carried out to wash-out, amount of water is 10 times of concentrated solution volume, obtain dialyzate, dialyzate is regulated to pH to 2 with dilute sulphuric acid, by adsorbing from storng-acid cation exchange resin 732 posts, adsorb the saturated rear ammoniacal liquor with pH10 the L-Leu on resin column is carried out to desorb, obtain stripping liquid 14kg altogether.After ultrafiltrated permeation liquid and stripping liquid are mixed through organic nanofiltration membrane decolouring desalination, adopt polyether sulfone filtering film, retaining molecular weight 800Da, pressure 1.0MPa, 40 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring and product in dope is carried out to wash-out obtains organic membrane destainer 150kg altogether, L-Leu content 40mg/L in destainer, transparence 87.4%, destainer is cooled to concentrated solution to 18 DEG C after evaporation concentration certain multiple and carries out decrease temperature crystalline, after crystal drying, obtain L-Leu crude product, L-Leu purity 93.3%, yield 96.0%.
Embodiment 7
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 50nm, filter pressure 0.2MPa, crossflow velocity 4m/s, after concentrating 10 times, fermented liquid obtains ultrafiltrated permeation liquid, add wash water to the middle stream of ultrafiltration dope (pH is 6～7) again L-Leu in dope and filter cake is carried out to wash-out (it is 8 that the middle sodium hydroxide that adds water makes pH), amount of water is 10 times of concentrated solution volume, obtain dialyzate, dialyzate is regulated to pH to 2 with dilute sulphuric acid, by adsorbing from storng-acid cation exchange resin 732 posts, resin demand is 5kg, adsorb the saturated rear ammoniacal liquor with pH10 the L-Leu on resin column is carried out to desorb, obtain stripping liquid 14kg altogether, completing whole adsorption-desorption manipulation require lives again resin 2 times.After ultrafiltrated permeation liquid and stripping liquid are mixed through organic nanofiltration membrane decolouring desalination, adopt polyether sulfone filtering film, retaining molecular weight 800Da, pressure 1.0MPa, 40 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring and product in dope is carried out to wash-out obtains organic membrane destainer 150kg altogether, L-Leu content 40mg/L in destainer, transparence 87.4%, destainer is cooled to concentrated solution to 18 DEG C after evaporation concentration certain multiple and carries out decrease temperature crystalline, after crystal drying, obtain L-Leu crude product, L-Leu purity 93.1%, yield 97.0%.Can find out according to embodiment 6 and embodiment 7, by adopting alkalescence washing when the dialysis operation, can change amino acid whose character, it is more easily seen through by ultra-filtration membrane, yield is improved.
Embodiment 8
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 50nm, filter pressure 0.2MPa, crossflow velocity 4m/s, after concentrating 10 times, fermented liquid obtains ultrafiltrated permeation liquid, add wash water to the middle stream of ultrafiltration dope (pH is 6～7) again L-Leu in dope and filter cake is carried out to wash-out (it is 8 that the middle sodium hydroxide that adds water makes pH), amount of water is 10 times of concentrated solution volume, obtain dialyzate, dialyzate is regulated to pH to 2 with dilute sulphuric acid, by adsorbing from storng-acid cation exchange resin 732 posts, resin demand is 5kg, adsorb the saturated rear ammoniacal liquor with pH10 the L-Leu on resin column is carried out to desorb, obtain stripping liquid 14kg altogether, completing whole adsorption-desorption manipulation require lives again resin 2 times.By after ultrafiltrated permeation liquid and stripping liquid mixing, after adding carbonic acid gas to make pH be 6, again through organic nanofiltration membrane decolouring desalination, adopt polyether sulfone filtering film, retaining molecular weight 800Da, pressure 1.0MPa, 40 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring and product in dope is carried out to wash-out obtains organic membrane destainer 150kg altogether, L-Leu content 40mg/L in destainer, transparence 87.4%, destainer is cooled to concentrated solution to 18 DEG C after evaporation concentration certain multiple and carries out decrease temperature crystalline, after crystal drying, obtain L-Leu crude product, L-Leu purity 95.5%, yield 97.2%.Can find out according to embodiment 7 and embodiment 8, by carrying out in the process of nanofiltration desalination, by making feed liquid acidifying, can improve the rejection of inorganic salt, product purity is further improved, and carbonic acid gas can be got rid of after heating evaporation from material, can not affected product purity.
Reference examples
Get 60 DEG C of high-temperature sterilization secondary fermentation liquid 150kg, after mixing through ceramic super-filtering film filtering and concentrating, the mean pore size of ceramic super-filtering film is 50nm, filter pressure 0.2MPa, crossflow velocity 4m/s, after concentrating 10 times, fermented liquid obtains ultrafiltrated permeation liquid, regulate pH to 2 to ultrafiltration dope with dilute sulphuric acid again, by adsorbing from storng-acid cation exchange resin 732 posts, resin demand 5kg, adsorb the saturated rear ammoniacal liquor with pH10 the L-Leu on resin column is carried out to desorb, obtain stripping liquid 160kg altogether, complete whole adsorption-desorption manipulation require resin is lived again 9 times.After stripping liquid is mixed through polyethersulfone reverse osmosis decolouring desalination, pressure 2.0MPa, 40 DEG C of temperature, the decolouring later stage adds pure water in dope to decolouring and product in dope is carried out to wash-out obtains organic membrane concentrated solution 40kg altogether, L-Leu content 9g/L in concentrated solution is cooled to concentrated solution 18 DEG C by concentrated solution and carries out decrease temperature crystalline after evaporation concentration certain multiple, obtains L-Leu crude product after crystal drying, L-Leu purity 92.4%, yield 94.0%.Can find out with embodiment 7 contrast, adopt ultrafiltration-dialysis-can significantly reduce ion-exchange resins use regeneration frequency and consumption from friendships-nanofiltration technique, and the product yield obtaining and purity are also better than reference examples.
Claims (10)
- The leucic extracting method of 1.L-, is characterized in that, comprises the steps:, by L-Leu fermented liquid ultrafiltration membrance filter, to obtain the first filtrate; Then the first filtrate is sent into separation circuit; Described separation circuit refers to and filters by first nanofiltration membrane, then nanofiltration permeate is obtained to L-Leu by after concentrated, crystallization.
- 2. the extracting method of L-Leu according to claim 1, is characterized in that: after ultrafiltration membrance filter, water carries out dialysis to the filter cake of ultra-filtration membrane, obtains the second filtrate, then the second filtrate is sent into described separation circuit.
- 3. the extracting method of L-Leu according to claim 2, is characterized in that: in described dialysis operation, amount of water is 5～15 times of concentrated liquid measure.
- 4. the extracting method of L-Leu according to claim 3, is characterized in that: the pH regulator of the water adding when dialysis is 8～9.
- 5. the extracting method of L-Leu according to claim 2, is characterized in that: the second filtrate being need to be after ion exchange resin carries out Adsorption and desorption processing, then sends into described separation circuit; Described ion exchange resin refers to storng-acid cation exchange resin.
- 6. according to the extracting method of L-Leu claimed in claim 5, it is characterized in that: described storng-acid cation exchange resin refers to 732, WA-2, HD-8, JK008 or HZ014.
- 7. the extracting method of L-Leu according to claim 1, is characterized in that: the molecular weight cut-off of described nanofiltration membrane is 500～1000Da.
- 8. the extracting method of L-Leu according to claim 1, is characterized in that: the material of described nanofiltration membrane is rhodia based polymer, polymeric amide, SPSF, polyethersulfone, polysulfones, polyacrylonitrile, polyester, polyimide and vinyl polymer; Preferred, polyethers sulfone or polysulfones.
- 9. the extracting method of L-Leu according to claim 1, is characterized in that: in ultrafiltration membrance filter process, working pressure is 0.1～0.3MPa; Crossflow velocity is 2～5m/s; Cycles of concentration is 5～20 times.
- 10. the extracting method of L-Leu according to claim 1, is characterized in that: during nanofiltration membrane is filtered, filter pressure is 0.5～1.5MPa, and filtration temperature is 30～45 DEG C.
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