CN103641766B - From zymotic fluid, extract continuously the method for L-Trp - Google Patents
From zymotic fluid, extract continuously the method for L-Trp Download PDFInfo
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- CN103641766B CN103641766B CN201310692894.0A CN201310692894A CN103641766B CN 103641766 B CN103641766 B CN 103641766B CN 201310692894 A CN201310692894 A CN 201310692894A CN 103641766 B CN103641766 B CN 103641766B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
- C07D209/18—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D209/20—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals substituted additionally by nitrogen atoms, e.g. tryptophane
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
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Abstract
The invention discloses a kind of method of extracting continuously L-Trp from zymotic fluid, it comprises the following steps: step 1, and zymotic fluid uses hydrochloric acid adjust pH to 3.8 to 4.5 in fermentation tank, and heat inactivation forms feed liquid; Step 2, feed liquid enters high speed tube centrifuge, and removal thalline and the insoluble protein of separating out form centrifugal efflux; Step 3, centrifugal efflux is removed most of pigment with 3000 daltonian films, and the efflux after decolouring enters 200 daltonian tubular type NF membrane system nanofiltrations to the micro-muddy formation concentrate of feed liquid; Step 4, concentrate is by the rapid decrease temperature crystalline of tube type mold, and crystal solution obtains tryptophan crude product after Separation of Solid and Liquid; Step 5, dissolves tryptophan crude product with hot pure water, add hydrochloric acid adjust pH to 4.0 to 5.0 hydrotropies; After activated carbon decolorizing, white tryptophan fine work is dried and obtained to same employing step 2, to the round-robin method processing of step 4. The present invention has improved quality and the yield of product.
Description
Technical field
The present invention relates to a kind of extraction, particularly relate to a kind of method of extracting continuously L-Trp from zymotic fluid.
Background technology
Tryptophan has been widely used in the aspects such as medicine, food, feed, and fermentation method production technology has greatly improved, and carriesThe technology of getting also improves to some extent and improves, and steams but also rest at present traditional decompression heating in the concentrated link of zymotic fluid post processingSend out stage of anhydrating, even if partial monopoly has adopted the processing of film system, also just do at the beginning of concentrated early stage densely with nanofiltration, then proceed toTraditional decompression heating evaporation anhydrates to crystallization, until concentration is complete. Processing again of mother liquor is also to adopt same traditionMethod.
The patent No. is that the Chinese patent of CN101717360A discloses and removes thalline and albumen with milipore filter, activated carbon decolorizing,The concentrated tryptophan of separating out of evaporimeter. Can make part tryptophan become pigment as traditional heating is concentrated, increase on the one hand de-The intractability of look, has also reduced the yield of tryptophan on the other hand.
The patent No. is that the Chinese patent of CN201010158401.1 discloses and removes thalline and albumen with ceramic membrane, and milipore filter removesPartial pigment, nanofiltration concentrates to obtain concentrate, then reduced pressure concentration. This patent still adopts traditional heating concentrated in the concentrated later stage, there is the variable color of reduced pressure concentration when heating tryptophan in method, yield is declined and the problem of Quality Down.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of method of extracting continuously L-Trp from zymotic fluid, itsGet rid of traditional evaporation and concentration by circulation nanofiltration decrease temperature crystalline method, improved quality and the yield of product.
The present invention solves above-mentioned technical problem by following technical proposals: a kind of L-that extracts continuously from zymotic fluidThe method of tryptophan, is characterized in that, it comprises the following steps:
Step 1, zymotic fluid uses hydrochloric acid adjust pH to 3.8 to 4.5 in fermentation tank, and heat inactivation forms feed liquid;
Step 2, feed liquid enters high speed tube centrifuge, and removal thalline and the insoluble protein of separating out form centrifugal outflowLiquid;
Step 3, centrifugal efflux is removed most of pigment with 3000 daltonian films, and the efflux after decolouring enters 200The nanofiltration of daltonian tubular type NF membrane system is to the micro-muddy formation concentrate of feed liquid;
Step 4, concentrate is by the rapid decrease temperature crystalline of tube type mold, and crystal solution obtains tryptophan after Separation of Solid and LiquidCrude product; The tubular type NF membrane system that is back to isolated mother liquor circulates concentrated again, until feed liquid processing finishes by the gross;
Step 5, dissolves tryptophan crude product with hot pure water, add hydrochloric acid adjust pH to 4.0 to 5.0 hydrotropies; De-through active carbonAfter look, white tryptophan fine work is dried and obtained to same employing step 2, to the round-robin method processing of step 4.
Preferably, described tubular type NF membrane system held temperature is 45 DEG C.
Preferably, the number of times of the activated carbon decolorizing of described step 5 is twice.
Preferably, described mother liquor mixes post-drying with wheat bran and is used as animal feed additive.
Positive progressive effect of the present invention is: the present invention utilizes high temperature nanofiltration to anhydrate and in conjunction with cooling, utilizes tryptophanWhen nanofiltration, easily separate out the characteristic of water, adopt tube type mold crystallization, by Separation of Solid and Liquid, tryptophan is separated, mother liquor is got back toNanofiltration system again concentrated circulation the continuous crystallization of mode, isolate crystal, recrystallization, isolation of crystalline again, reach conventional methodThe effect of concentrated and crystallization, but mother liquor amount of the present invention is few, and yield is high.
Detailed description of the invention
Following examples are only for the present invention is described, but the present invention is not subject to the restriction of these embodiment.
The method that the present invention extracts continuously L-Trp from zymotic fluid comprises the following steps:
Step 1, zymotic fluid uses hydrochloric acid adjust pH to 3.8 to 4.5 in fermentation tank, and heat inactivation forms feed liquid;
Step 2, feed liquid enters high speed tube centrifuge, and removal thalline and the insoluble protein of separating out form centrifugal outflowLiquid;
Step 3, centrifugal efflux is removed most of pigment with 3000 daltonian films, and the efflux after decolouring enters 200The nanofiltration of daltonian tubular type NF membrane system is to the micro-muddy formation concentrate of feed liquid;
Step 4, concentrate is by the rapid decrease temperature crystalline of tube type mold, and crystal solution obtains tryptophan after Separation of Solid and LiquidCrude product; The tubular type NF membrane system that is back to isolated mother liquor circulates concentrated again, until feed liquid processing finishes by the gross;
Step 5, dissolves tryptophan crude product with hot pure water, add hydrochloric acid adjust pH to 4.0 to 5.0 hydrotropies; De-through active carbonAdopt that nanofiltration is concentrated equally after look, the round-robin method place of decrease temperature crystalline, Separation of Solid and Liquid, nanofiltration concentrated (step 2 is to step 4)Reason, dries and obtains white tryptophan fine work.
Embodiment 1
The method that the present invention (embodiment 1) extracts continuously L-Trp from zymotic fluid comprises the following steps:
The L-Trp zymotic fluid that is 40.18g/L by 1000L concentration is used hydrochloric acid adjust pH to 4.1 in fermentation tank, and addsHot deactivation forms feed liquid;
Feed liquid enters high speed tube centrifuge, and removal thalline and the insoluble protein of separating out form centrifugal efflux;
Centrifugal efflux is removed most of pigment with 3000 daltonian films, and the efflux after decolouring is daltonian with 200The nanofiltration of tubular type NF membrane system is to the micro-muddy formation concentrate of feed liquid, and tubular type NF membrane system held temperature is 45 DEG C;
Concentrate forms crystal solution by tube type mold decrease temperature crystalline, and it is thick that crystal solution obtains tryptophan through Separation of Solid and LiquidProduct, isolated mother liquor is back to tubular type NF membrane system and again circulates, until the whole processing of feed liquid finish by the gross.
The aforementioned hot pure water of tryptophan crude product 96.80kg obtaining is dissolved, add hydrochloric acid adjust pH to 4.3 hydrotropies. Add workProperty charcoal 1.0kg, 70 DEG C of decolourings 1 hour, cross elimination charcoal, filtrate adds active carbon 0.5kg again and carries out secondary decolourization, 60 DEG C of decolourings 1 are littleTime, after coarse filtration, filter to obtain subdiaphanous smart filtrate by 0.45 μ m filter membrane essence again. Essence filtrate by 200 daltonian tubular type NF membrane isSystem is concentrated, and nanofiltration system holding temperature is 52 DEG C, and nanofiltration is micro-muddy to feed liquid, and concentrate is through tube type mold cryosel cooling crystallization,Magma liquid obtains tryptophan wet product through Separation of Solid and Liquid, and mother liquor is back to NF membrane system and again circulates, and it is pure white that drying obtains color and lusterTryptophan fine work 35.16kg, light transmittance is 98.53%; Specific rotation is-30.8 °; Meet standards of pharmacopoeia.
The present embodiment is 87.51% to the extract yield of tryptophan in zymotic fluid.
Embodiment 2
The method that the present invention (embodiment 2) extracts continuously L-Trp from zymotic fluid comprises the following steps:
Hydrochloric acid adjust pH to 4.2 for the L-Trp zymotic fluid that is 39.97g/L by 2500L concentration, and heat inactivation formsFeed liquid;
Feed liquid is pressed into high speed tube centrifuge with compressed air, removes thalline and the insoluble protein of separating out forms centrifugalEfflux;
Centrifugal efflux enters the feed liquid basin of decolouring film system, removes most of pigment with 3000 daltonian films, takes offEfflux after look enters the feed liquid basin of 200 daltonian tubular type NF membrane systems, and nanofiltration is to the micro-muddy formation concentrate of feed liquid,Tubular type NF membrane system held temperature is 45 DEG C;
Concentrate forms crystal solution by tube type mold decrease temperature crystalline, enters pocket type automatic centrifuge. Centrifugal discharge Dry SackPropylhomoserin crude product. Mother liquor is back to NF membrane system and again circulates, until the whole processing of feed liquid finish. After can mixing with wheat bran, dries in mother liquorThe dry animal feed additive of being used as;
Tryptophan crude product 232.7kg is dissolved with hot pure water 1830L, and hydrochloric acid adjust pH is to 4.5 hydrotropies. Add active carbon2.5kg, 70 DEG C are incubated 1 hour, cross elimination charcoal, obtain yellow filtrate. Filtrate adds active carbon 1.2kg again, and to carry out secondary charcoal de-, 60 DEG CDecolour 1 hour, after coarse filtration, again with 0.45 μ m filter membrane essence filter, obtain subdiaphanous smart filtrate. Essence filtrate is by 200 daltonian tubular typesNF membrane system is concentrated to slightly muddy, cryosel cooling crystallization, and brilliant pulp separation, mother liquor returns and pumps into NF membrane system and again circulate, straightFinish to the whole processing of feed liquid. The centrifugal material drying of washing after crystalline substance obtains the tryptophan fine work 87.7kg that color and luster is pure white. Light transmittanceBe 99.20%; Specific rotation is-31.2 °, meets standards of pharmacopoeia.
It is 87.77% that tryptophan in zymotic fluid is calculated to extract yield.
Embodiment 3
The method that the present invention (embodiment 3) extracts continuously L-Trp from zymotic fluid comprises the following steps:
The L-Trp zymotic fluid that is 40.02g/L by 2000L concentration is used hydrochloric acid adjust pH to 3.8 in fermentation tank, and addsHot deactivation forms feed liquid;
Feed liquid enters high speed tube centrifuge, and removal thalline and the insoluble protein of separating out form centrifugal efflux;
Centrifugal efflux is removed most of pigment with 3000 daltonian films, and the efflux after decolouring is daltonian with 200The nanofiltration of tubular type NF membrane system is to the micro-muddy formation concentrate of feed liquid, and tubular type NF membrane system held temperature is 45 DEG C;
Concentrate forms crystal solution by tube type mold decrease temperature crystalline, and it is thick that crystal solution obtains tryptophan through Separation of Solid and LiquidProduct, isolated mother liquor is back to tubular type NF membrane system and again circulates, until the whole processing of feed liquid finish by the gross.
The aforementioned hot pure water of tryptophan crude product 100.6kg obtaining is dissolved, add hydrochloric acid adjust pH to 4.0 hydrotropies. Add workProperty charcoal 2.0kg, 70 DEG C of decolourings 1 hour, cross elimination charcoal, filtrate adds active carbon 1.5kg again and carries out secondary decolourization, 60 DEG C of decolourings 1 are littleTime, after coarse filtration, filter to obtain subdiaphanous smart filtrate by 0.45 μ m filter membrane essence again. Essence filtrate by 200 daltonian tubular type NF membrane isSystem is concentrated, and nanofiltration system holding temperature is 52 DEG C, and nanofiltration is micro-muddy to feed liquid, and concentrate is through tube type mold cryosel cooling crystallization,Magma liquid obtains tryptophan wet product through Separation of Solid and Liquid, and mother liquor is back to NF membrane system and again circulates, and it is pure white that drying obtains color and lusterTryptophan fine work 40.35kg, light transmittance is 98.72%; Specific rotation is-30.6 °; Meet standards of pharmacopoeia.
The present embodiment is 87.62% to the extract yield of tryptophan in zymotic fluid.
Embodiment 4
The method that the present invention (embodiment 4) extracts continuously L-Trp from zymotic fluid comprises the following steps:
The L-Trp zymotic fluid that is 41.65g/L by 1500L concentration is used hydrochloric acid adjust pH to 4.5 in fermentation tank, and addsHot deactivation forms feed liquid;
Feed liquid enters high speed tube centrifuge, and removal thalline and the insoluble protein of separating out form centrifugal efflux;
Centrifugal efflux is removed most of pigment with 3000 daltonian films, and the efflux after decolouring is daltonian with 200The nanofiltration of tubular type NF membrane system is to the micro-muddy formation concentrate of feed liquid, and tubular type NF membrane system held temperature is 45 DEG C;
Concentrate forms crystal solution by tube type mold decrease temperature crystalline, and it is thick that crystal solution obtains tryptophan through Separation of Solid and LiquidProduct, isolated mother liquor is back to tubular type NF membrane system and again circulates, until the whole processing of feed liquid finish by the gross.
The aforementioned hot pure water of tryptophan crude product 103.2kg obtaining is dissolved, add hydrochloric acid adjust pH to 4.5 hydrotropies. Add workProperty charcoal 4.0kg, 70 DEG C of decolourings 1 hour, cross elimination charcoal, filtrate adds active carbon 0.2kg again and carries out secondary decolourization, 60 DEG C of decolourings 1 are littleTime, after coarse filtration, filter to obtain subdiaphanous smart filtrate by 0.45 μ m filter membrane essence again. Essence filtrate by 200 daltonian tubular type NF membrane isSystem is concentrated, and nanofiltration system holding temperature is 52 DEG C, and nanofiltration is micro-muddy to feed liquid, and concentrate is through tube type mold cryosel cooling crystallization,Magma liquid obtains tryptophan wet product through Separation of Solid and Liquid, and mother liquor is back to NF membrane system and again circulates, and it is pure white that drying obtains color and lusterTryptophan fine work 45.35kg, light transmittance is 98.12%; Specific rotation is-30.2 °; Meet standards of pharmacopoeia.
The present embodiment is 87.43% to the extract yield of tryptophan in zymotic fluid.
The present invention divides with concentrate → decrease temperature crystalline → solid-liquid of concentrate → decrease temperature crystalline → Separation of Solid and Liquid → nanofiltration of high temperature nanofiltrationFrom continuous circulation processing method replace traditional heating under diminished pressure, mild condition, technical process does not have the negative of localized hyperthermiaImpact, nanofiltration concentrates when anhydrating and has also removed small molecular weight impurity and inorganic ion etc., and the speed of anhydrating is also than decompression heatingThe evaporation speed of anhydrating is fast, and energy consumption is low, and the quality of tryptophan is improved significantly, and yield is corresponding increasing also. Root of the present inventionWhen concentrated according to tryptophan nanofiltration, water outlet is very fast, viscosity is little, the characteristic of difficult stifled NF membrane, in the concentrated film processing of high-temperature tubular nanofiltrationTime can concentrate feed is directly dense slightly muddy to material, fast cooling post crystallization is good, according to this feature, whole process using nanofiltration is denseContracting, simple to operate, mild condition, without hot-spot, yield is slightly high compared with conventional method, and quality is good, through a small amount of activated carbon decolorizing twoInferior, then, cooling concentrated by continous way film system, separate, dry, get final product to such an extent that meet the tryptophan product of standards of pharmacopoeia.
The invention is not restricted to these disclosed embodiment, the present invention is by described soverlay technique scheme scope, Yi JiquanVarious distortion and the equivalence of profit claimed range change, and are not departing under the prerequisite of technical solution of the present invention, to the present inventionAny amendment that the those skilled in the art that do easily realize or improvement all belong to the present invention's scope required for protection.
Claims (1)
1. a method of extracting continuously L-Trp from zymotic fluid, is characterized in that, it comprises the following steps:
Step 1, zymotic fluid uses hydrochloric acid adjust pH to 3.8 to 4.5 in fermentation tank, and heat inactivation forms feed liquid;
Step 2, feed liquid enters high speed tube centrifuge, and removal thalline and the insoluble protein of separating out form centrifugal efflux;
Step 3, centrifugal efflux is removed most of pigment with 3000 daltonian films, and the efflux after decolouring enters 200 Dao ErThe tubular type NF membrane system nanofiltration of pausing is to the micro-muddy formation concentrate of feed liquid;
Step 4, concentrate is by the rapid decrease temperature crystalline of tube type mold, and crystal solution obtains tryptophan crude product after Separation of Solid and Liquid;The tubular type NF membrane system that is back to isolated mother liquor circulates concentrated again, until feed liquid processing finishes by the gross;
Step 5, dissolves tryptophan crude product with hot pure water, add hydrochloric acid adjust pH to 4.0 to 5.0 hydrotropies; After activated carbon decolorizingL-Trp is dried and obtained to same employing step 2, to the method processing of step 4;
Described tubular type NF membrane system held temperature is 45 DEG C;
The number of times of the activated carbon decolorizing of described step 5 is twice.
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| CN103864667B (en) * | 2014-03-24 | 2016-04-13 | 无锡荣丰生物工程有限公司 | L-Trp economic benefits and social benefits consecutive evaporation deamination technique |
| 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 |
| CN110041242A (en) * | 2019-05-27 | 2019-07-23 | 山东银丰生物科技有限公司 | A kind of extraction process technology and equipment of tryptophan fermentation liquid |
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