CN1061217A - From fermented liquid, extract the method for L-Methionin - Google Patents
From fermented liquid, extract the method for L-Methionin Download PDFInfo
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Abstract
The present invention relates to from fermented liquid, extract the method for L-Methionin, it mainly comprises with the organic or inorganic flocculation agent removes mycelium in the fermented liquid and protein and removes mineral ion in the fermented liquid with oxalic acid, spent ion exchange resin carries out the absorption and the desorb thereof of L-Methionin, with sorbent material stripping liquid is decoloured, concentrate destainer and the crystallization from concentrated solution of L-Methionin is separated out, it is characterized in that except that oxalic acid, the agent of removing mineral ion from fermented liquid also comprises tripoly phosphate sodium STPP, and described ion-exchange is degree of crosslinking greater than 12% resin, and described sorbent material is a granular active carbon.
Description
The present invention relates to the extracting method of Methionin, more particularly, relate to the method for from fermented liquid, extracting L-Methionin.
As everyone knows, Methionin is one of eight kinds of indispensable amino acids of humans and animals nutrition, and it as foodstuff additive, can be increased Nutritive value of food, improves the utilization ratio of food proteins; And with it as fodder additives, then can promote the growth of animal; In addition, Methionin also can be used for pharmaceutically, and it is good that the moriamin-s that L-Methionin and other amino acid are mixed preparation has nutrition, and pyrogen-free matter and allergin are safe in utilization, advantages such as long preservative period.Therefore, for the production of Methionin, people had carried out a large amount of research already.
At present, Methionin mostly with amylum hydrolysate of the sugar or molasses as raw material, utilize fermentation using bacteria to produce, and adopt ion exchange method to extract, usually with the direct upper prop of fermented liquid (without filtering), through concentrate, decolouring, twice crystallization and obtain finished product.Technical process is long, yield lower (65-75%), and raw materials consumption is many, the hydrochloric acid that will consume in the existing extraction process, ammoniacal liquor, the expense of ion exchange resin etc. generally will account for Feng 50%(such as the star of total cost, " the scientific and technological communication of fermenting " 13(2), P.99-102(1984), so adopting suitable method to extract Methionin is necessary to reduce production costs.
Usually, a complete ion exchange extraction method should comprise following five steps:
1. fermentation liquor pretreatment;
2. ion exchange resin adsorbs L-Methionin;
3. select the adsorbed L-Methionin of certain strippant wash-out for use;
4. to the elutriant processing of decolouring;
5. concentrate destainer and with its crystallization.
The purpose of fermentation liquor pretreatment is to remove existing mycelium in the fermented liquid, and the protein of remnants and some mineral ions are (as Ca in the substratum
2+, Mg
2+Deng).Cameron in No. 6407 south african patent (1970), C.﹠amp; Weiss, K. have disclosed with non-ionic type high molecular cellulose precipitation agent and have come separation of mycelial.People such as Yan Xikang (" microbiology circular " 14(2), P54-58(1987)) disclosed with non-ionic polyacrylamide and cationic polyacrylamide and removed mycelium and protein in the fermented liquid, all obtained effect preferably.
At present, the method for removing mineral ion generally is to add oxalic acid in fermented liquid, and like this, calcium ion and oxalic acid reaction can generate calcium oxalate precipitation.But because the solubleness of magnesium oxalate is bigger, so this method can not be removed the magnesium ion in the fermented liquid effectively.
In general, can directly carry out ion-exchange absorption through pretreated fermented liquid.At present, the ion exchange resin that general employing degree of crosslinking is 7-8% in the production adsorbs Methionin, and this resin has higher adsorptive capacity for Methionin, but its selectivity is bad, can adsorb more pigment simultaneously, gives the refining difficulty of having brought of Methionin.
In addition, the decolouring of L-Methionin generally adopts powdered active carbon as pigment adsorbent at present.This method is based on gac and has bigger specific surface area, and it can behind the elimination gac, just can obtain the L-lysine solution through decolouring by physisorption and with impurity absorption such as pigments on the surface.This method operation is simpler, but the inventor adopts powdered active carbon to decolour by discovering, can cause L-Methionin greater loss, and the regeneration of powdered active carbon is difficult, can not reuse, and makes production cost higher.And owing to adopt plate-and-frame filter press just can remove powdered active carbon, so can not produce continuously.
Last step of L-Methionin extraction process is that the L-Methionin crystallization after the decolouring is obtained finished product.Crystallization method commonly used at present has two kinds: (1) adds a certain amount of ethanol in the L lysine HCL solution of decolouring back (or concentrating the back), because the hydrochloride of L-Methionin is insoluble to ethanol, (Muradyan, A.G., SU.891,643), crystallization yield increases with the increase of ethanol dosage, when the ethanol dosage is 1 times of L lysine HCL solution, crystallization yield can reach more than 90%, but along with the increase of ethanol dosage, the problem that crystal purity descends then occurs, can not satisfy the demand.In addition, reclaiming the alcoholic acid process need increases some equipment and makes and be difficult to use in scale operation.(2) in the aqueous solution, obtain the product of L lysine HCL by the continuous quadratic crystallization, the detailed process of this method is: will be concentrated into 23Be ° through the L lysine HCL of decolouring, then its pH is transferred to 4.9, about 10 hours of crystallization at a certain temperature filters to isolate crystal; The mother liquor reconcentration is pressed the said process recrystallize once again to about 23Be °, and the purity of the crystalline product that so obtains also can meet the requirements.The latter is used on producing widely, but the inventor finds the best pH when pH=4.9 is not crystallization by a large amount of research, and crystallization can not obtain maximum crystallization yield under this pH condition.
The purpose of this invention is to provide a kind of method of from fermented liquid, extracting L-Methionin, adopt this method, can high quality and high yield ground obtain required L-Methionin product, and production cost is reduced.
The objective of the invention is to finish: a kind of method of from fermented liquid, extracting L-Methionin by following design, comprise with the organic or inorganic flocculation agent and remove mycelium in the fermented liquid and protein and remove mineral ion in the fermented liquid with oxalic acid, spent ion exchange resin carries out the absorption and the desorb of L-Methionin, with sorbent material stripping liquid is decoloured, concentrate destainer and the crystallization from concentrated solution of L-Methionin is separated out, it is characterized in that except that oxalic acid the agent of removing mineral ion from fermented liquid also comprises tripoly phosphate sodium STPP; And described ion exchange resin is the interlinkage degree greater than 12% resin, and described sorbent material is a granular active carbon.
Described organic floculant is polyacrylamide or chitosan, and described inorganic flocculating agent is aluminium chlorohydroxide or CaCl
2Or Na
2HPO
4The add-on of described oxalic acid is a 0.05-0.2%(weight), be preferably 0.08-0.12%(weight), the add-on of described tripoly phosphate sodium STPP is a 0.1-0.3%(weight), be preferably 0.12-0.22%(weight).
The degree of crosslinking of described ion exchange resin is preferably 14-16%.The particle diameter of described granular active carbon is the 10-70 order, is preferably the 20-30 order.
Above-mentioned sorbent material also comprises the condensed type weak-acid ion exchange resin, and granular active carbon and condensed type weak-acid ion exchange resin are used in combination and decolour.Described concentration through spissated destainer is 20-30Be °, is preferably 22-25Be °.And adopting the mode of regulating pH that the crystallization from concentrated solution of L-Methionin is separated out, described pH the best is 5.5.
Advantage of the present invention and effect are tangible.Since the fermentation pre-treatment in oxalic acid and tripoly phosphate sodium STPP are used in combination, they respectively with Ca
2+And Mg
2+(Ca
2+And Mg
2+Be the main inorganic impurity ion that influences ion-exchange in the L-lysine fermentation liquor) in conjunction with and generate the calcium oxalate precipitation of indissoluble and the MgNa of solubility
3P
3O
10Complex compound, and can not participate in ion-exchange.Remove by filter post precipitation, just can obtain the ferment filtrate that impurity level significantly reduces, help ion-exchange subsequently.In addition, owing to select the ion exchange resin of high-crosslinking-degree for use, this resinoid selectivity is higher than the resin of employed medium degree of crosslinking at present, the pigment of absorption seldom, the quality of the L-lysine solution of institute's wash-out can be improved, help follow-up decolouring step greatly, also help improving the final product quality of L-Methionin.
In addition, the present invention adopts granular active carbon as sorbent material, and granular active carbon can be loaded in the exchange column and decolour, like this, production is carried out continuously, shortened the production cycle, and powdered active carbon does not reach this requirement, because if powdered active carbon is filled in the post, fluid resistance is too big, operation can't be carried out, so, when adopting powdered active carbon, must remove gac with plate-and-frame filter press as sorbent material, can not serialization production.In case of necessity, also condensed type weak-acid ion exchange resin and granular active carbon tandem of the present invention can be decoloured, and obtain better decolorizing effect.
In addition, the present inventor is by discovering in a large number, make the crystallization of L-Methionin about pH regulator to 5.5 with the L-lysine solution after concentrating, can obtain maximum L-Methionin crystallization yield, than produce at present the crystallization yield of the L-Methionin that obtained under the situation of general pH=4.9 improve about 1.5-2%, and the purity of resulting L-Methionin can reach more than 99%, and the quality of product is improved, relatively, cost has just reduced.
Below, will advantage of the present invention and effect be described in further detail by specific examples.
Example 1: the pre-treatment of fermented liquid
In the L-fermenting lysine, contain a considerable amount of organic and inorganic impurities, organic impurity mainly is mycelium and protein, and inorganic impurity mainly is Ca
2+And Mg
2+These impurity can influence the carrying out of ion-exchange, so it is very necessary to remove impurity as far as possible that fermented liquid is carried out pre-treatment.
Organic impurity in the L-lysine fermentation liquor generally adopts polymeric flocculant to remove, wherein with the effect that adopts polyacrylamide flocculant for well, operate with the ratio that adds the polyacrylamide flocculant of 1.2-1.5ml 0.2% in the 100ml L-lysine fermentation liquor, at suitable pH(such as pH=6.0) under, resulting supernatant liquor light absorption value less (being that foreign matter content is few), and the amount of throw out is more.
Ca in the L-lysine fermentation liquor
2+Can remove by adding the calcium oxalate precipitation that oxalic acid forms indissoluble volumes such as (, " production of antibiotics technology ", Chemical Industry Press, (1982)) Wu Hangyan, and Mg
2+Then can be by method of the present invention, promptly add tripoly phosphate sodium STPP and form the complex compound of tripoly phosphate sodium STPP magnesium.The add-on of oxalic acid is a 0.05-0.2%(weight), be preferably 0.08-0.12%(weight), the add-on of tripoly phosphate sodium STPP is a 0.1-0.3%(weight), be preferably 0.12-0.22%(weight).
In general, can be earlier with the Ca in oxalic acid and the tripoly phosphate sodium STPP removal fermented liquid
2+And Mg
2+Ion, then with the pH regulator to 6.0 of solution, add polyacrylamide flocculant, in case of necessity, also can add a little wilkinite (it can improve flocculation agent for protein and mycelial flocculating effect), add a little diatomite again and help filter, suction filtration can obtain through pretreated L-lysine fermentation liquor then.
Example 2: to the selection of the ion exchange resin of absorption L-Methionin, absorption and elution process
The ion exchange resin that is adopted in the L-Methionin production at present is that degree of crosslinking is the resin of 7-8%, this resinoid is bigger to the adsorptive capacity of L-Methionin, but simultaneously, it also has bigger adsorptive power to pigment, promptly bad to the adsorption selectivity of L-Methionin, thus, brought difficulty with making with extra care for L-Lysine Decolorant subsequently.
The present inventor selects the resin of various degree of crosslinking to carry out it to the absorption of L-Methionin and the test of wash-out.(in 10 * 100mm) ion exchange column, with the feeding through pretreated fermented liquid of pH regulator to 2.0, the space velocity s.v. of feeding is 1/20min then at first the ion exchange resin of a certain amount of (as 10ml) to be drained and is packed into certain specification
-1About, saturated to adsorbing (detecting with 731 type spectrophotometers) uses excessive 7%NH then
4The OH wash-out, s.v.=1/40min
-1Adopting down, formula I calculates dynamic total exchange capacity and desorb (wash-out) percentage of various resins for L-Methionin respectively with (II):
Q= ((C
1-C
2)V′)/(V) (Ⅰ)
Wherein: the total exchanging container of Q=(mg/ml resin)
C
1The starting point concentration of=L-lysine fermentation liquor (mg/ml)
C
2=raffinate concentration (mg/ml)
The volume (ml) of the L-lysine fermentation liquor of V '=passed through
The volume of the resin that V=packed into (ml)
Wherein: the mean concns of C=stripping liquid (mg/ml)
V " the volume of=stripping liquid (ml)
The meaning of other symbol is the same.
Resulting the results are shown in the following table 1.
The various resins of table 1. are for the dynamic exchange capacity and the desorption efficiency thereof of L-Methionin
Resin | Total exchange capacity Q(mg/ml) | Desorption efficiency (%) |
The Dowex1 Dowex2 Dowex4 732(degree of cross linking is 7) Dowex8 Dowex12 strong acid 1 * 14.5 Dowex16 | 37.0 49.4 67.5 115.0 118.0 136.0 149.0 137.0 | 98.7 99.5 99.6 98.4 98.3 97.6 98.9 96.5 |
The concentration of the L-lysine fermentation liquor that is * adopted is 2.79%, and the concentration of stoste and stripping liquid is all filled photometric detection with 731 type branches.
Data can see that Dowex 12,1 * 14.5 from table, and the total exchange capacity and the desorption efficiency of the resin of 16 3 kinds of high interlinkage degree of Dowex are all better, and in experiment, the inventor finds, the NH with 7%
4After the OH desorb, the color restoration original state of these three kinds of resins, the color of other resin is then darker, and this may be that pigment can not enter resin inside owing to these three kinds of cross-linkage of resin height.The resin of this proof high-crosslinking-degree has selectivity preferably for L-Methionin.
The decolouring of example 3:L-Methionin stripping liquid is handled
Producing the upward general decolouring of adopting powdered active carbon to carry out L-Methionin stripping liquid at present handles, when adopting this method to decolour, the loss of L-Methionin is bigger, and powdered active carbon is difficult to regeneration, caused production cost higher, again owing to must adopt Plate Filtration to remove powdered active carbon, so, can not carry out operate continuously.
In view of gac itself has bigger specific surface area, it is a kind of good pigment adsorbent, the present inventor is by a large amount of experiments, discovery can adopt granular gac to replace pulverous gac, like this, just can realize continuous operation, (compare because granular active carbon can being filled in the post decolours with powdered active carbon, the fluid resistance of granular active carbon is much smaller), the particle diameter of granular active carbon is generally the 10-70 order, is preferably the 20-30 order.
The 10ml granular active carbon is packed in the post of φ 20 * 200mm, feed the stripping liquid of L-Methionin, s.v. is 1/50min
-1, obtain the adsorption curve of a tears shaped activated carbon, as shown in Figure 1 for pigment.As can see from Figure 1, granular active carbon has bigger adsorptive power for pigment.
Use the adsorbed pigment of 0.5N NaOH desorb again, obtain result (s.v.=1/50min shown in Figure 2
-1Can observe from Fig. 2, the elution peak of pigment is concentrated, no conditions of streaking, and the regeneration problem of this explanation granular active carbon can be solved.Therefore, production cost can correspondingly reduce.
In case of necessity, condensed type weakly acidic resin and granular active carbon tandem can be carried out the decolouring of L-lysine solution, like this, can obtain better decolorizing effect, further improve the quality of product.The regeneration of condensed type weakly acidic resin also can be adopted the NaOH about 0.5N.
Concentrating and crystallization of embodiment 4:L-Methionin
To place through the L-Methionin dope that decolouring is handled and be evaporated to certain degree in the rotatory evaporator, be generally 20-30Be °, be preferably 22-25Be °.
Get the beaker of 12 150ml, respectively put into 25ml through spissated L-lysine solution, under 10 ℃, the pH with HCl regulates the L-Methionin concentrated solution in a plurality of beakers respectively observes the crystalline quantity that produces under different pH conditions.
The result shows, resulting amount in the condition of resulting L-Methionin crystal amount greater than pH=4.9 under the condition of pH=5.5.Can see that thus pH5.5 is the best pH of L-Methionin crystalline.Under this pH condition, the solubleness minimum of L-Methionin (hydrochloride) in the aqueous solution.
Experiment finds, the crystallization yield during pH5.5 on average exceeds 1.5% during than pH4.9.And resulting crystalline purity can reach 99.8% under the pH5.5 condition.
Crystallization is better under pH5.5, is valid.Because the pK of Methionin
1=2.18, pK
2=8.9, and at pH=1/2 (PK
1+ PK
2)=1/2 (2.18+8.90)=5.54 o'clock, the concentration maximum of Methionin mono-hydrochloric salts.
Concrete method is: will transfer to 5.5 through the pH of spissated L-lysine solution, and then at a certain temperature, be preferably 10 ℃ of following crystallizations, and after about 10 hours, filter to isolate crystal.The crystallization yield of resultant L-Methionin can reach about 92%, meets the requirement of generation, and need not secondary crystal, thereby has further reduced production cost and alleviated labour intensity.
Example 5: adopt method of the present invention to produce the process of L-Methionin
Respectively in 0.1% and 0.17%(weight) ratio oxalic acid and tripoly phosphate sodium STPP are added in the L-lysine fermentation liquor, regulate the pH to 6.0 of fermented liquid then, optionally add 0.2%(weight) wilkinite, under agitation condition, with 1-1.5ml/100ml fermentation 0.2%(weight) wilkinite, under agitation condition, ratio adding 0.2%(weight with the 1-1.5ml/100ml fermented liquid) polyacrylamide flocculant, after treating that a large amount of throw outs occur, add 1.0%(weight again) diatomite as flocculating aids, suction filtration under the vacuum tightness of 600mmHg discards filter residue.
Then, will transfer to 2.0 through the pH of pretreated L-lysine solution with 6NHCl.Place stand-by.
1 * 14.5 NH packs in the ion exchange column of φ a 85 * 1200mm
4Resin (low price of this resin can be used for scale operation), resin device is about 3000ml.Then above-mentioned solution upper prop is carried out ion-exchange absorption (s.v.=1/40min
-1); When outflow concentration when the concentration of fermented liquid is identical during with upper prop, stop to feed fermented liquid.Then, use salt-free water rinse resin, use 7.0% NH again
4OH carries out desorb (SV=1/90min
-1); When the pH of elutriant reaches 8.5 left and right sides, begin to collect elutriant, the amount of L-Methionin stops to collect elutriant seldom the time in the liquid that flows out to post.Then, stripping liquid is placed the rotatory evaporator evaporation remove deammoniation.About pH regulator to 4.5, place stand-by again with stripping liquid.
Other gets the ion exchange column of φ a 40 * 600mm, toward the 500ml granular active carbon (particle diameter is the 30-60 order) of wherein packing into, then above-mentioned elutriant is fed the (s.v.=1/20min that decolours
-1) (look particular case, also available condensed type weakly acidic resin is (as 122
#Resin) further decolouring), collected destainer is placed rotatory evaporator, at 60 ℃, be concentrated into about 23Be ° under the 740mmHg, under same temperature with the pH regulator to 5.5 of concentrated solution, naturally cooling under agitation, be preferably in 10 ℃ of following crystallizations, after 4 hours, isolate crystal, be the L-Methionin finished product that is obtained.
The result that the method that relatively adopts method of the present invention to produce L-Methionin and employing prior art is produced L-Methionin finds that method of the present invention can obtain better result, and concrete data are listed in the following table 3 and table 4.
The method of table 3 prior art is produced L-Methionin
The experiment number | 1 | 2 | 3 |
Fermentation liquid measure (ml) | 1200 | 1200 | 1200 |
Fermented liquid concentration (%) | 2.80 | 2.80 | 2.80 |
The amount (g) of L-Methionin (hydrochloride) in the fermented liquid | 336.00 | 336.00 | 336.00 |
Crystallization obtains the amount (g) of L-Methionin (hydrochloride) | 269.47 | 267.12 | 271.15 |
Yield (%) | 80.2 | 79.5 | 80.7 |
Average yield (%) | 80.1 |
Table 4 method of the present invention is produced L-Methionin
The experiment number | 1 | 2 | 3 |
Fermentation liquid measure (ml) | 1200 | 1200 | 1200 |
Fermented liquid concentration (%) | 2.80 | 2.80 | 2.80 |
The amount (g) of L-Methionin (hydrochloride) in the fermented liquid | 336.00 | 336.00 | 336.00 |
Crystallization obtains the amount (g) of L-Methionin (hydrochloride) | 286.67 | 284.36 | 285.62 |
Yield (%) | 85.3 | 84.6 | 85.0 |
Average yield (%) | 85.0 |
Therefore advantage of the present invention and effect are tangible.
Above-mentioned example of the present invention is appreciated that in aim of the present invention and scope just in order to help the person skilled in art to understand and to implement the present invention, can makes multiple modification.
Claims (11)
1, a kind of method of extracting L-Methionin from fermented liquid comprises that removing mycelium in the fermented liquid and protein and oxalic acid with the organic or inorganic flocculation agent removes mineral ion in the fermented liquid; Spent ion exchange resin carries out the absorption and the desorb thereof of L-Methionin; With sorbent material stripping liquid is decoloured, concentrate destainer and the crystallization from concentrated solution of L-Methionin is separated out, it is characterized in that except that oxalic acid, the agent of removing mineral ion from fermented liquid also comprises tripoly phosphate sodium STPP, and described ion exchange resin is degree of crosslinking greater than 12% resin, and described sorbent material is a granular active carbon.
2, the method for claim 1 is characterized in that described organic floculant is polyacrylamide or chitosan, and described inorganic flocculating agent is aluminium chlorohydroxide or CaCl
2Or Na
2HPO
4
3, the method for claim 1, the add-on that it is characterized in that described oxalic acid are 0.05-0.2%(weight), be preferably 0.08-0.12%(weight).
4, the method for claim 1, the add-on that it is characterized in that described tripoly phosphate sodium STPP are 0.1-0.3%(weight), be preferably 0.12-0.22%(weight).
5, the method for claim 1 is characterized in that the degree of crosslinking of described ion exchange resin is preferably 14-16%.
6, the method for claim 1 is characterized in that the particle diameter of described granular active carbon is the 10-70 order, is preferably the 20-30 order.
7,, it is characterized in that described sorbent material also comprises the condensed type weakly acidic resin as claim 1 or 6 described methods.
8, the method for claim 1, it is characterized in that described granular active carbon and condensed type weakly acidic resin the series connection decolour.
9, the method for claim 1, it is characterized in that the concentration of spissated destainer be 20-30Be °, be preferably 22-25Be °.
10, the method for claim 1 is characterized in that adopting the mode of regulating pH that the crystallization from concentrated solution of L-Methionin is separated out.
11, as claim 1 or 10 described methods, it is characterized in that making the L-Methionin best pH that crystallization is separated out from concentrated solution is 5.5.
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Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US2894026A (en) * | 1956-08-01 | 1959-07-07 | Du Pont | Purification of lysine |
US3036125A (en) * | 1959-06-02 | 1962-05-22 | Pfizer & Co C | Process for purifying lysine |
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CN1041824C (en) | 1999-01-27 |
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