CN104561202A - Preparation method and technological system for enzymatically synthesizing N(2)-L-alanyl-L-glutamine - Google Patents
Preparation method and technological system for enzymatically synthesizing N(2)-L-alanyl-L-glutamine Download PDFInfo
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Abstract
The invention provides a preparation method and technological system for enzymatically synthesizing N(2)-L-alanyl-L-glutamine. The preparation method comprises the steps of synthesis of L-alanine methyl ester, synthesis of N(2)-L-alanyl-L-glutamine, inactivation of enzymes, filtration, preparation of a crude product I of N(2)-L-alanyl-L-glutamine and preparation of a crude product II of N(2)-L-alanyl-L-glutamine. The technological system also comprises a production line formed by an L-alanine methyl ester synthesis kettle (1), an L-alanine methyl ester storage tank (2), an N(2)-L-alanyl-L-glutamine synthesis reaction kettle (3), an enzyme storage tank (4), a storage tank (5), a resin column (6) and mother liquor recovery tanks (7). The preparation method and the technological system have the technical effects that impurities are effectively removed, thus improving the purity of a finished product; the requirements for equipment are simple; the yield is high, after-treatment is simple to carry out, and the cost is low.
Description
Technical field
The present invention relates to a kind of method of synthesizing glutamine dipeptide, especially adopt the glutamine dipeptide biosynthetic means of efficient biological enzyme synthetic technology, specifically a kind of preparation method of Enzyme catalyzed synthesis glutamine dipeptide and process system.
Background technology
N (2)-L-Ala-L-Glu amine (also known as Dipeptiven) belongs to amino acid whose derivative, is called for short glutamine dipeptide, is mainly used in body-care industry, is the ideal substitute of the abundantest amino acid glutamine of body burden.
The physiological function very important due to glutamine and pharmacological action, make its application in parenteral nutrition be subject to the general attention of people.But because its solubleness is low, and unstable in solution, under the condition of heat sterilization, generate poisonous burnt glutaric acid and ammonia, so not containing glutamine in commodity amino acid preparation.Only be translated into stable derivative to work to human body.
For glutamine, the solubleness of glutamine dipeptide is 20 times of glu famine, also very stable in storage and heat sterilization, directly can be prepared into infusion preparation for clinical.Namely glutamine dipeptide resolves into glutamine rapidly after entering human body and plays a role.Research proves, glutamine dipeptide is broken down into its composition amino acid in vivo very soon, transformation period is very short, a small amount of dipeptides can only be detected in blood, the dipeptides of trace is only had to discharge from urine, illustrate that glutamine dipeptide effectively can be utilized and can not gather in blood, avoid issuable pharmacology and physiological impairment.Healthy human body long-term intravenous instillation glutamine dipeptide, without any side effect and untoward reaction, does not affect normal renal function.
CN101659691A and CN1786019A discloses a kind of chemosynthesis and industrialized preparing process (following route) of glutamine dipeptide, but the method Reactive Synthesis route is long, and environmental pollution is large, and process costs is high.
Existing enzymatic clarification is prepared in glutamine dipeptide subsequent technique, can be mingled with the enzyme in reaction solution in crude product, is difficult to remove, and affects purity and the quality of glutamine dipeptide.
Summary of the invention
In order to solve the problem, first object of the present invention is the processing method providing a kind of Enzyme catalyzed synthesis glutamine dipeptide.
The present invention's second object is to provide a kind of process system of Enzyme catalyzed synthesis glutamine dipeptide.
There is provided prepared by the method that the biosynthesizing of glutamine dipeptide provided by the invention adopts biological enzyme to adopt the patent No. to prepare for the biological enzyme of glutamine dipeptide in 201410670694.X.
Technical scheme of the present invention: the biosynthetic reaction formula of biological enzyme glutamine dipeptide is as follows:
The biosynthetic method of described biological enzyme glutamine dipeptide provided by the invention, is specified as following steps:
(1) synthesis of ALANINE methyl esters salt
A. methyl alcohol is joined in reactor, and be cooled to 0-10 DEG C;
B. thionyl chloride is added dropwise in reactor at 0-10 DEG C;
C. thionyl chloride drips completely and joins in reactor by disposable for L-Ala, insulated and stirred 30min, is more slowly warming up to 25-30 DEG C, stirs 1h, then is warming up to 45-50 DEG C of stirring 2h;
D. 50 DEG C of decompressions evaporate most solvent, obtain ALANINE methyl esters salt;
(2) synthesis of glutamine dipeptide
A. the ALANINE methyl ester hydrochloride prepared by above-mentioned steps soluble in water and be cooled to 0-10 DEG C for subsequent use;
B. water to be joined in reactor and to be cooled to 5-10 DEG C;
C. L-glutaminate is joined in reactor, and adjust pH to 8.8-9.2 with ammoniacal liquor at such a temperature, the enzyme of 50%-60% enzyme amount is joined in reaction system;
D. be slowly added dropwise in reactor at 5-10 DEG C by the ALANINE methyl ester hydrochloride aqueous solution, adjust pH to make pH maintain 8.9-9.0 with ammoniacal liquor, add in reactor by above-mentioned enzyme, stirring reaction generates glutamine dipeptide reaction solution.
(3) inactivation of enzyme
A. methanol solution is added;
B. above-mentioned reaction solution is warming up to 80-95 DEG C;
C. filter after adding the gac stirring 20-30min of 5-7 ‰;
(4) filter
A. above-mentioned reaction solution positive resin acidic solution absorption;
B. resin is washed with water;
C. use ammoniacal liquor wash-out resin, collecting washings is glutamine dipeptide crude product solution;
(5) preparation of glutamine dipeptide crude product one
A. glutamine dipeptide wet product above-mentioned steps obtained and water add in dissolution kettle, control temperature 30-35 DEG C stirring and dissolving; Add ammoniacal liquor and adjust pH to 7.3-7.5;
B., under 45-50 DEG C of outer temperature, be concentrated in above-mentioned solution and have solid to occur, be cooled to 30-35 DEG C;
C. be slowly added dropwise to by methyl alcohol in above-mentioned reaction system, the mass ratio of methyl alcohol and above-mentioned nanofiltration concentrated solution is 5-6:1;
D. drip after stirring 20-30min completely and be cooled to 5-10 DEG C, centrifugal after insulated and stirred 1h; E. glutamine dipeptide crude product is obtained with the methyl alcohol of 20% quantity of methyl alcohol used and the mixing solutions rinsing of water;
(6) preparation of glutamine dipeptide crude product two
A. dissolve: the glutamine dipeptide crude product one above-mentioned steps obtained and the water of 1.0 times of weight add in dissolution kettle, 30-35 DEG C of stirring and dissolving; Ammoniacal liquor adjusts pH to 7.2-7.3;
B. solution 50 DEG C of outer temperature being concentrated in solution has solid to occur, is cooled to 30-35 DEG C;
C. methyl alcohol is slowly added dropwise in reaction system;
D. drip after stirring 30min completely and be cooled to 5-10 DEG C, centrifugal after insulated and stirred 1h;
E. glutamine dipeptide crude product two is obtained with the methyl alcohol of 20% quantity of methyl alcohol used and the mixing solutions rinsing of water;
(7) glutamine dipeptide is refining
A. dissolve: the glutamine dipeptide crude product two above-mentioned steps obtained and water add in dissolution kettle, 40 DEG C of stirring and dissolving;
B. decolour: reactor adds the gac of 5-7 ‰, stir 30min, filter;
C. above-mentioned filtered liquid is dry, obtain glutamine dipeptide finished product.
The invention provides the process system of Enzyme catalyzed synthesis glutamine dipeptide, comprise production line, this production line comprises ALANINE methyl esters synthesis reactor, ALANINE methyl esters basin, the third paddy synthesis reaction vessel, enzyme basin, basin, resin column, disposing mother liquor tank; Described ALANINE methyl esters synthesis reactor is positioned at the upstream of described ALANINE methyl esters basin, connected by pipeline therebetween, the upstream of the third described paddy synthesis reaction vessel is connected with described ALANINE methyl esters basin, described enzyme basin, ammoniacal liquor feed pot pipeline respectively, and the downstream of this third paddy synthesis reaction vessel is connected with the opening for feed pipeline of described basin; The upstream of described resin column is connected with the discharge port of described basin, acid tube road, ammoniacal liquor pipeline, waterpipe respectively; The downstream of described resin column is connected with pipeline between described disposing mother liquor tank.
As preferably, described basin has three, is connected in series by pipeline head and the tail.
As preferably, described production line has three, and between the resin column in every bar production line, pipeline is connected in parallel.
As preferably, in described resin column, there is positive resin packing layer.
Advantageous Effects of the present invention:
The invention provides a kind of preparation method of Enzyme catalyzed synthesis glutamine dipeptide, the present invention utilizes biological enzyme to synthesize glutamine dipeptide; Through adding methanol solution, heat up heating, enzyme deactivation unnecessary in reaction solution, protein denaturation is flocculated, through activated carbon filtration, the enzyme of removing flocculation, carries out adsorbing, adding water removal chlorion with resin column, then elutes glutamine dipeptide crude product with ammoniacal liquor, with methyl alcohol and elutriation crystalline substance after concentrated, obtain highly purified glutamine dipeptide.
The preparation method of Enzyme catalyzed synthesis glutamine dipeptide provided by the invention, synthetic route is simple, and raw material is cheaply easy to get, and equipment requirements is simple, with enzyme amount few (0.2%), in the reaction times short (2-3 hour), removes unreacted enzyme, improves the purity of finished product; Simple to equipment requirements; Yield is high, and aftertreatment is simple, and product purity is high; Cost is low, has high economic worth and the market competitiveness.
The invention provides a kind of process system of Enzyme catalyzed synthesis glutamine dipeptide, comprise three production lines, resin column in each production line is parallel with one another, and three production lines can be produced simultaneously, also can produce by technique alternate cycles, article three, the resin column absorption of production line, wash-out, regenerative process cycle alternation carry out, the acid tail washings of absorption is used for another group resin regeneration, and the 3rd group of wash-out collects product, this process system economy, efficient, environmental protection, tail washings can effectively fully recycle, and quantity discharged is little.Disposing mother liquor tank has three, head and the tail series connection, the disposing mother liquor tank being arranged in downstream is collected and the mother liquor concentrated turns back to the disposing mother liquor tank of upstream, the glutamine dipeptide crude product mother solution concentration of collecting in the disposing mother liquor tank be directly connected with resin column is the highest, resin column can be returned and carry out the higher glutamine dipeptide solution of absorb-elute acquisition concentration, refine further and obtain target compound.
Accompanying drawing explanation
Fig. 1 is schematic diagram of the present invention.
Embodiment
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.
Embodiment 1:
180kg methyl alcohol is joined in 1000L reactor, and is cooled to 0 DEG C; 78.6kg thionyl chloride is added dropwise in reactor at 0 DEG C; Thionyl chloride drips completely and joins in reactor by disposable for 52.5kgL-L-Ala, insulated and stirred 30min, is more slowly warming up to 25 DEG C, stirs 1h, then is warming up to 45 DEG C of stirring 2h; 50 DEG C of decompressions evaporate most solvent, obtain 90kgL-alanine methyl ester salt;
Embodiment 2:
180kg methyl alcohol is joined in 1000L reactor, and is cooled to 10 DEG C; 78.6kg thionyl chloride is added dropwise in reactor at 10 DEG C; Thionyl chloride drips completely and joins in reactor by disposable for Methionin, insulated and stirred 30min, is more slowly warming up to 30 DEG C, stirs 1h, then is warming up to 50 DEG C of stirring 2h; 50 DEG C of decompressions evaporate most solvent, obtain 82L-alanine methyl ester salt;
Embodiment 3:
180kg methyl alcohol is joined in 1000L reactor, and is cooled to 5 DEG C; 78.6kg thionyl chloride is added dropwise in reactor at 5 DEG C; Thionyl chloride drips completely and joins in reactor by disposable for Methionin, insulated and stirred 30min, is more slowly warming up to 28 DEG C, stirs 1h, then is warming up to 48 DEG C of stirring 2h; 50 DEG C of decompressions evaporate most solvent, obtain 88kgL-alanine methyl ester salt;
Embodiment 4:
The synthetic method of ALANINE methyl esters is the same.58g L-glutaminate (397mmol) is added in 400mL water, be cooled to 10 DEG C, adjust pH to 9.0 with ammonia soln, 1.5ml enzyme liquid is joined in reaction system, then 82g ALANINE methyl ester hydrochloride (590mmol) (is dissolved in 100mL by the ALANINE methyl esters aqueous solution
waterin) be added dropwise in reaction system, drip ammoniacal liquor simultaneously, make pH remain on 8.9; Measure enzyme liquid concentration in reaction process, timing repeatedly adds enzyme liquid, and when pH is constant, reaction terminates; Add methyl alcohol (reaction solution 20%) and be heated to 60 DEG C of inactivation 30min, filter after adding the gac stirring 20min of 5 ‰ and remove biological enzyme throw out; Reaction solution concentrated removing methyl alcohol, with positive resin absorption reaction liquid, after washing removing chloride with water, product is resolved again with 1% ammoniacal liquor, the part of collecting product concentrates, and controlling outer temperature is 50 DEG C, is concentrated in solution and has solid to occur, carry out being cooled to 30 DEG C, add 300ml methanol aqueous solution, drip completely and stir 30min, be cooled to 5 DEG C of filtrations, crystallization 4 hours, solid obtains glutamine dipeptide crude product one 60g at 50 DEG C of decompression dryings.
60g glutamine dipeptide crude product one is dissolved in 60mL water, adds 300mL methyl alcohol, slow cooling to 10 DEG C, crystallization 4h, obtains glutamine dipeptide crude product two, is dissolved in water, in 40 DEG C, stir 30min, add 5 ‰ gacs and stir 30min, filter, centrifuging, 51g glutamine dipeptide sterling is pulverized to obtain in oven dry, yield 85%, purity 99.0%.Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 two annex V D).
Embodiment 5:
The synthetic method of ALANINE methyl esters is the same.58g L-glutaminate (397mmol) is added in 400mL water, be cooled to 10 DEG C, adjust pH to 8.8 with ammonia soln, 1.5ml enzyme liquid is joined in reaction system, then 82g ALANINE methyl ester hydrochloride (590mmol) (is dissolved in 100mL by the ALANINE methyl esters aqueous solution
waterin) be added dropwise in reaction system, drip ammoniacal liquor simultaneously, make pH remain on 9.0; Measure enzyme liquid concentration in reaction process, timing repeatedly adds enzyme liquid, and when pH is constant, reaction terminates; Add methyl alcohol (20% of reaction solution) and be heated to 70 DEG C of inactivation 30min, filter after adding the gac stirring 30min of 7 ‰ and remove biological enzyme throw out; Reaction solution concentrated removing methyl alcohol, with positive resin absorption reaction liquid, after washing removing chloride with water, product is resolved again with 1% ammoniacal liquor, the part of collecting product concentrates, and controlling outer temperature is 50 DEG C, is concentrated in solution and has solid to occur, carry out being cooled to 35 DEG C, add 300ml methanol aqueous solution, drip completely and stir 30min, be cooled to 5 DEG C of filtrations, crystallization 4 hours, solid obtains glutamine dipeptide crude product one 60.5g at 50 DEG C of decompression dryings.
60.5g glutamine dipeptide crude product one is dissolved in 60.5mL water, adds 300mL methyl alcohol, slow cooling to 8 DEG C, crystallization 4h, obtains glutamine dipeptide crude product two, is dissolved in water, in 40 DEG C, stir 30min, add 7 ‰ gacs and stir 30min, filter, centrifuging, 51.5g glutamine dipeptide sterling is pulverized to obtain in oven dry, yield 86%, purity 99.2%.Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 two annex V D).
Embodiment 6:
The synthetic method of ALANINE methyl esters is the same.58g L-glutaminate (397mmol) is added in 400mL water, be cooled to 10 DEG C, adjust pH to 8.8 with ammonia soln, 1.5ml enzyme liquid is joined in reaction system, then 82g ALANINE methyl ester hydrochloride (590mmol) (is dissolved in 100mL by the ALANINE methyl esters aqueous solution
waterin) be added dropwise in reaction system, drip ammoniacal liquor simultaneously, make pH remain on 9.0; Measure enzyme liquid concentration in reaction process, timing repeatedly adds enzyme liquid, and when pH is constant, reaction terminates; Add methyl alcohol (20% of reaction solution) and be heated to 80 DEG C of inactivation 30min, filter after adding the gac stirring 30min of 7 ‰ and remove biological enzyme throw out; Reaction solution concentrated removing methyl alcohol, with positive resin absorption reaction liquid, after washing removing chloride with water, product is resolved again with 1% ammoniacal liquor, the part of collecting product concentrates, and controlling outer temperature is 50 DEG C, is concentrated in solution and has solid to occur, carry out being cooled to 30 DEG C, add 300ml methanol aqueous solution, drip completely and stir 30min, be cooled to 5 DEG C of filtrations, crystallization 4 hours, solid obtains glutamine dipeptide crude product one 61.5g at 50 DEG C of decompression dryings.
61.5g glutamine dipeptide crude product one is dissolved in 61.5mL water, adds 300mL methyl alcohol, slow cooling to 5 DEG C, crystallization 4h, obtains glutamine dipeptide crude product two, is dissolved in water, in 40 DEG C, stir 30min, add 7 ‰ gacs and stir 30min, filter, centrifuging, 51.8g glutamine dipeptide sterling is pulverized to obtain in oven dry, yield 87%, purity 99.5%.Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 two annex V D).
Embodiment 7:
The synthetic method of ALANINE methyl esters is the same.58g L-glutaminate (397mmol) is added in 400mL water, be cooled to 10 DEG C, adjust pH to 9.2 with ammonia soln, 1.5ml enzyme liquid is joined in reaction system, then 82g ALANINE methyl ester hydrochloride (590mmol) (is dissolved in 100mL by the ALANINE methyl esters aqueous solution
waterin be added dropwise in reaction system, drip ammoniacal liquor simultaneously, make pH remain on 9.0; Measure enzyme liquid concentration in reaction process, timing repeatedly adds enzyme liquid, and when pH is constant, reaction terminates; Add methyl alcohol (20% of reaction solution) and be heated to 80 DEG C of inactivation 30min, filter after adding the gac stirring 30min of 7 ‰ and remove biological enzyme throw out; Reaction solution concentrated removing methyl alcohol, with positive resin absorption reaction liquid, after washing removing chloride with water, product is resolved again with 1% ammoniacal liquor, the part of collecting product concentrates, and controlling outer temperature is 50 DEG C, is concentrated in solution and has solid to occur, carry out being cooled to 40 DEG C, add 300ml methanol aqueous solution, drip completely and stir 30min, be cooled to 8 DEG C of filtrations, crystallization 4 hours, solid obtains glutamine dipeptide crude product one 59.6g at 50 DEG C of decompression dryings.
59.6g glutamine dipeptide crude product one is dissolved in 59mL water, adds 300mL methyl alcohol, slow cooling to 5 DEG C, crystallization 4h, obtains glutamine dipeptide crude product two, is dissolved in water, in 40 DEG C, stir 30min, add 7 ‰ gacs and stir 30min, filter, centrifuging, 51.2g glutamine dipeptide sterling is pulverized to obtain in oven dry, yield 83%, purity 99.0%.Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 two annex V D).
As Fig. 1, a kind of process system of Enzyme catalyzed synthesis glutamine dipeptide, comprises production line, and this production line comprises ALANINE methyl esters synthesis reactor 1, ALANINE methyl esters basin 2, third paddy synthesis reaction vessel 3, enzyme basin 4, basin 5, resin column 6, disposing mother liquor tank 7; Described ALANINE methyl esters synthesis reactor 1 is positioned at the upstream of described ALANINE methyl esters basin 2, connected by pipeline therebetween, the upstream of the third described paddy synthesis reaction vessel 3 is connected with described ALANINE methyl esters basin 2, described enzyme basin 4, ammoniacal liquor feed pot pipeline respectively, and the downstream of this third paddy synthesis reaction vessel 3 is connected with the opening for feed pipeline of described basin 5; The upstream of described resin column 5 is connected with the discharge port of described basin 5, acid tube road, ammoniacal liquor pipeline, waterpipe respectively; The downstream of described resin column 5 is connected with pipeline between described disposing mother liquor tank 7.
Basin 4 in the present embodiment has three, is connected in series by pipeline head and the tail; Described production line has three, and between the resin column 6 in every bar production line, pipeline is connected in parallel; Positive resin packing layer is had in described resin column 6.
The synthesis of ALANINE methyl esters is carried out in ALANINE methyl esters synthesis reactor 1, then ALANINE methyl esters basin 2 is collected in, when starting to synthesize glutamine dipeptide, ALANINE methyl esters basin 2, enzyme basin 4 and ammoniacal liquor pipeline feed in raw material to the third paddy synthesis reaction vessel 3 simultaneously, through reaction, reaction solution is collected in basin 5, then heats flocculation, carry out enzyme deactivation, the solution after filtration enters that resin column carries out adsorbing, wash-out.
The invention provides a kind of process system of Enzyme catalyzed synthesis glutamine dipeptide, comprise three production lines, resin column in each production line is parallel with one another, and three production lines can be produced simultaneously, also can produce by technique alternate cycles, article three, the resin column absorption of production line, wash-out, regenerative process cycle alternation carry out, the acid tail washings of absorption is used for another group resin regeneration, and the 3rd group of wash-out collects product, this process system economy, efficient, environmental protection, tail washings can effectively fully recycle, and quantity discharged is little.Disposing mother liquor tank has three, head and the tail series connection, the disposing mother liquor tank being arranged in downstream is collected and the mother liquor concentrated turns back to the disposing mother liquor tank of upstream, the glutamine dipeptide crude product mother solution concentration of collecting in the disposing mother liquor tank be directly connected with resin column is the highest, resin column can be returned and carry out the higher glutamine dipeptide solution of absorb-elute acquisition concentration, refine further and obtain target compound.
Present specification describes some embodiments, but should be understood that those skilled in the art pass through to read this specification sheets and can know the various improvement not deviating from the spirit and scope of the present invention.Therefore, these other embodiments also should be included within the scope of appended claims.
Claims (6)
1. a preparation method for Enzyme catalyzed synthesis glutamine dipeptide, is characterized in that comprising the following steps:
(1) synthesis of ALANINE methyl esters salt
A. methyl alcohol is joined in reactor, and be cooled to 0-10 DEG C;
B. thionyl chloride is added dropwise in reactor at 0-10 DEG C;
C. thionyl chloride drips completely and joins in reactor by disposable for L-Ala, insulated and stirred 20min-30min, is more slowly warming up to 25-30 DEG C, stirs 30min-1h, then is warming up to 45-50 DEG C of stirring 1-2h;
D. 50 DEG C of decompressions evaporate most solvent, obtain ALANINE methyl esters salt;
(2) synthesis of glutamine dipeptide
A. the ALANINE methyl ester hydrochloride prepared by above-mentioned steps soluble in water and be cooled to 0-10 DEG C for subsequent use;
B. water to be joined in reactor and to be cooled to 5-10 DEG C;
C. L-glutaminate is joined in reactor, and adjust pH to 8.8-9.2 with ammoniacal liquor at such a temperature, the enzyme of 50%-60% enzyme amount is joined in reaction system;
D. be slowly added dropwise in reactor at 5-10 DEG C by the ALANINE methyl ester hydrochloride aqueous solution, adjust pH to make pH maintain 8.9-9.0 with ammoniacal liquor, add in reactor by above-mentioned enzyme, stirring reaction generates glutamine dipeptide reaction solution;
(3) inactivation of enzyme
A. methanol solution is added;
B. above-mentioned reaction solution is warming up to 60-80 DEG C;
C. filter after adding the gac stirring 20-30min of 5-7 ‰;
(4) filter
A. above-mentioned reaction solution positive resin acidic solution absorption;
B. resin is washed with water;
C. use ammoniacal liquor wash-out resin, collecting washings is glutamine dipeptide crude product solution;
(5) preparation of glutamine dipeptide crude product one
A. glutamine dipeptide wet product above-mentioned steps obtained and water add in dissolution kettle, control temperature 30-35 DEG C stirring and dissolving; Add ammoniacal liquor and adjust pH to 7.3-7.5;
B., under 45-50 DEG C of outer temperature, be concentrated in above-mentioned solution and have solid to occur, be cooled to 30-35 DEG C;
C. be slowly added dropwise to by methyl alcohol in above-mentioned reaction system, the mass ratio of methyl alcohol and above-mentioned solution is 5-6:1;
D. drip after stirring 20-30min completely and be cooled to 5-10 DEG C, centrifugal after insulated and stirred 1h; E. glutamine dipeptide crude product one is obtained with the mixing solutions rinsing of methyl alcohol and water;
(6) preparation of glutamine dipeptide crude product two
A. dissolve: the glutamine dipeptide crude product one above-mentioned steps obtained and the water of 1.0 times of weight add in dissolution kettle, 30-35 DEG C of stirring and dissolving; Ammoniacal liquor adjusts pH to 7.2-7.3;
B. above-mentioned solution is kept 50 DEG C of outer temperature, be concentrated in solution and had solid to occur, be cooled to 30-35 DEG C;
C. methyl alcohol is slowly added dropwise in reaction system;
D. drip after stirring 30min completely and be cooled to 5-10 DEG C, centrifugal after crystallization insulated and stirred 1h;
E. glutamine dipeptide crude product two is obtained with the aqueous solution rinsing of methyl alcohol;
(7) glutamine dipeptide is refining
A. dissolve: the glutamine dipeptide crude product two above-mentioned steps obtained and water add in dissolution kettle, 40 DEG C of stirring and dissolving;
B. decolour: reactor adds the gac of 5-7 ‰, stir 30min, filter;
C. above-mentioned filtered liquid is dry, obtain glutamine dipeptide finished product.
2. the preparation method of Enzyme catalyzed synthesis glutamine dipeptide according to claim 1, is characterized in that, in the synthesis of step (2) glutamine dipeptide, described biological enzyme, ALANINE methyl esters, ammoniacal liquor evenly add reactor simultaneously; Described biological enzyme interval joins in reaction system all several times.
3. the process system of an Enzyme catalyzed synthesis glutamine dipeptide as claimed in claim 1, it is characterized by: comprise production line, this production line comprises ALANINE methyl esters synthesis reactor (1), ALANINE methyl esters basin (2), the third paddy synthesis reaction vessel (3), enzyme basin (4), basin (5), resin column (6), disposing mother liquor tank (7); Described ALANINE methyl esters synthesis reactor (1) is positioned at the upstream of described ALANINE methyl esters basin (2), connected by pipeline therebetween, the upstream of the third described paddy synthesis reaction vessel (3) is connected with described ALANINE methyl esters basin (2), described enzyme basin (4), ammoniacal liquor feed pot pipeline respectively, and the downstream of this third paddy synthesis reaction vessel (3) is connected with the opening for feed pipeline of described basin (5); The upstream of described resin column (5) is connected with the discharge port of described basin (5), acid tube road, ammoniacal liquor pipeline, waterpipe respectively; The downstream of described resin column (5) is connected with pipeline between described disposing mother liquor tank (7).
4. the process system of Enzyme catalyzed synthesis glutamine dipeptide according to claim 3, is characterized by: described basin (4) has three, is connected in series by pipeline head and the tail.
5. the process system of Enzyme catalyzed synthesis glutamine dipeptide according to claim 3, is characterized by: described production line has three, and between the resin column (6) in every bar production line, pipeline is connected in parallel.
6. the process system of Enzyme catalyzed synthesis glutamine dipeptide according to claim 3, is characterized by: described resin column has positive resin packing layer in (6).
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| CN106701869A (en) * | 2016-12-30 | 2017-05-24 | 山东辰龙药业有限公司 | Preparation method of N (2)-L-alanyl-L-glutamine |
| CN110407913A (en) * | 2018-06-11 | 2019-11-05 | 合肥川迪医药技术有限公司 | A kind of process for separation and purification of glutamine dipeptide |
| CN115160164A (en) * | 2022-05-06 | 2022-10-11 | 大连医诺生物股份有限公司 | Preparation method and application of composite alanine methyl ester hydrochloride |
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