CN104561202B - A kind of preparation method and technology system of Enzyme catalyzed synthesis glutamine dipeptide - Google Patents
A kind of preparation method and technology system of Enzyme catalyzed synthesis glutamine dipeptide Download PDFInfo
- Publication number
- CN104561202B CN104561202B CN201510064439.5A CN201510064439A CN104561202B CN 104561202 B CN104561202 B CN 104561202B CN 201510064439 A CN201510064439 A CN 201510064439A CN 104561202 B CN104561202 B CN 104561202B
- Authority
- CN
- China
- Prior art keywords
- glutamine dipeptide
- enzyme
- synthesis
- added
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The present invention provides a kind of preparation method and technology system of Enzyme catalyzed synthesis glutamine dipeptide, includes the synthesis of L methyl lactamines, the synthesis of glutamine dipeptide, the inactivation of enzyme, filtering, the preparation of glutamine dipeptide crude product one, the preparation of glutamine dipeptide crude product two.Present invention process system also includes L methyl lactamine synthesis reactors（1）, L methyl lactamine basins（2）, the third paddy synthesis reaction vessel（3）, enzyme basin（4）, basin（5）, resin column（6）, disposing mother liquor tank（7）The production line of composition.The present invention effectively goes the removal of impurity, improves the purity of finished product；It is simple to equipment requirement；High income, post processing is simple, and cost is low.
Description
Technical field
The present invention relates to a kind of method for synthesizing glutamine dipeptide, in particular by efficient biological enzyme synthetic technology
Glutamine dipeptide biological synthesis method, the specifically a kind of preparation method and process system of Enzyme catalyzed synthesis glutamine dipeptide.
Background technology
N (2)-L-Ala-L-Glu amine（Also known as alanyl glutamine）Belong to the derivative of amino acid, referred to as
Glutamine dipeptide, it is mainly used in body-care industry, is the ideal substitute of the most abundant amino acid glutamine of body burden.
Due to the particularly important physiological function of glutamine and pharmacological action, make its application in parenteral nutrition by people
Universal attention.But because its solubility is low, and it is unstable in solution, under conditions of heat sterilization, generation has
The burnt glutaric acid and ammonia of poison, so all not containing glutamine in commodity amino acid preparation.Only it is translated into stable derivative
Thing could work to human body.
For glutamine, the solubility of glutamine dipeptide is 20 times of glu famine, in storage and heat sterilization
In it is also very stable, can directly be prepared into infusion preparation be used for clinic.Glutamine dipeptide resolves into rapidly glutamy after entering human body
Amine and play a role.Research has shown that glutamine dipeptide is broken down into it and forms amino acid quickly in vivo, and half-life period is very short, blood
In be only able to detect a small amount of dipeptides, only micro dipeptides is discharged from urine, illustrates that glutamine dipeptide can be effectively utilized
And will not gather in blood, avoid issuable pharmacology and physiological impairment.Healthy human body long-term intravenous instillation third
Paddy dipeptides does not have any side effect and adverse reaction, does not influence normal renal function.
CN101659691A and CN1786019A discloses chemical synthesis and the industrialized preparing process of a kind of glutamine dipeptide
（Following route）, but this method reaction synthetic route length, environmental pollution is big, and process costs are high.
Existing enzymatic clarification is prepared in glutamine dipeptide subsequent technique, the enzyme that can be mingled with crude product in reaction solution, is very difficult to
Remove, influence the purity and quality of glutamine dipeptide.
The content of the invention
In order to solve the above problems, of the invention first purpose is to provide a kind of work of Enzyme catalyzed synthesis glutamine dipeptide
Process.
Second purpose of the invention is to provide a kind of process system of Enzyme catalyzed synthesis glutamine dipeptide.
The biosynthesis of glutamine dipeptide provided by the invention is used in Patent No. 201410670694.X using biology enzyme
The prepared offer of method prepared by the biology enzyme of glutamine dipeptide.
Technical scheme：The reaction equation of the biosynthesis of biological enzyme glutamine dipeptide is as follows：
The method of the biosynthesis of described biological enzyme glutamine dipeptide provided by the invention, is specified as following steps：
（1）The synthesis of ALANINE methyl esters salt
A. methanol is added in reactor, and is cooled to 0-10 DEG C；
B. thionyl chloride is added dropwise in reactor at 0-10 DEG C；
C. alanine is disposably added in reactor by thionyl chloride after being added dropwise completely, insulated and stirred 30min, then is delayed
It is slow to be warming up to 25-30 DEG C, 1h is stirred, then be warming up to 45-50 DEG C of stirring 2h；
D. 50 DEG C of decompressions evaporate most solvents, obtain ALANINE methyl esters salt；
（2）The synthesis of glutamine dipeptide
A. by above-mentioned steps prepare ALANINE methyl ester hydrochloride it is soluble in water and be cooled to 0-10 DEG C it is standby；
B. add water in reactor and be cooled to 5-10 DEG C；
C. Glu is added in reactor, and adjusts pH to 8.8-9.2 with ammoniacal liquor at such a temperature, by 50%-
The enzyme of 60% enzyme amount is added in reaction system；
D. the ALANINE methyl ester hydrochloride aqueous solution is slowly added dropwise in reactor at 5-10 DEG C, adjusts pH to make with ammoniacal liquor
PH maintains 8.9-9.0, and above-mentioned enzyme is added in reactor, stirring reaction generation glutamine dipeptide reaction solution.
（3）The inactivation of enzyme
A. methanol solution is added；
B. above-mentioned reaction solution is warming up to 80-95 DEG C；
C. filtered after adding 5-7 ‰ activated carbon stirring 20-30min；
（4）Filtering
A. above-mentioned reaction solution positive resin acid solution adsorbs；
B. resin is eluted with water；
C. resin is eluted with ammoniacal liquor, collection cleaning solution is glutamine dipeptide crude product solution；
（5）The preparation of glutamine dipeptide crude product one
A. the glutamine dipeptide wet product and water above-mentioned steps obtained is added in dissolution kettle, and control 30-35 DEG C of stirring of temperature is molten
Solution；Ammonification water adjusts pH to 7.3-7.5；
B. under 45-50 DEG C of outer temperature, being concentrated into above-mentioned solution has solid appearance, is cooled to 30-35 DEG C；
C. methanol is slowly added dropwise in above-mentioned reaction system, methanol and the mass ratio of above-mentioned nanofiltration concentrate are 5-6：
1；
D. 5-10 DEG C is cooled to after stirring 20-30min after being added dropwise completely, is centrifuged after insulated and stirred 1h；
E. the methanol of quantity of methyl alcohol and the mixed solution of water rinse to obtain glutamine dipeptide crude product used in 20%；
（6）The preparation of glutamine dipeptide crude product two
A. dissolve：The glutamine dipeptide crude product one and the water of 1.0 times of weight that above-mentioned steps are obtained are added in dissolution kettle, 30-
35 DEG C of stirring and dissolvings；Ammoniacal liquor adjusts pH to 7.2-7.3；
B. 50 DEG C of outer temperature of solution have been concentrated into solution has solid appearance, is cooled to 30-35 DEG C；
C. methanol is slowly added dropwise in reaction system；
D. 5-10 DEG C is cooled to after stirring 30min after being added dropwise completely, is centrifuged after insulated and stirred 1h；
E. the methanol of quantity of methyl alcohol and the mixed solution of water rinse to obtain glutamine dipeptide crude product two used in 20%；
（7）Glutamine dipeptide refines
A. dissolve：The glutamine dipeptide crude product two and water that above-mentioned steps are obtained are added in dissolution kettle, 40 DEG C of stirring and dissolvings；
B. decolourize：Reactor adds 5-7 ‰ activated carbon, stirs 30min, filtering；
C. above-mentioned filtered fluid is dried, obtains glutamine dipeptide finished product.
The present invention provides the process system of Enzyme catalyzed synthesis glutamine dipeptide, including production line, the production line include the ammonia of L- third
Sour methyl esters synthesis reactor, ALANINE methyl esters basin, the third paddy synthesis reaction vessel, enzyme basin, basin, resin column, disposing mother liquor tank；Institute
The ALANINE methyl esters synthesis reactor stated is located at the upstream of described ALANINE methyl esters basin, is connected therebetween by pipeline, described
The third paddy synthesis reaction vessel upstream respectively with the ALANINE methyl esters basin, the enzyme basin, ammoniacal liquor feed pot pipeline connect
Connect, the downstream of the third paddy synthesis reaction vessel is connected with the charging aperture pipeline of the basin；The upstream of the resin column respectively with institute
Discharging opening, acid tube road, ammoniacal liquor pipeline, the waterpipe for stating basin are connected；The downstream of described resin column and the disposing mother liquor
Pipeline connects between tank.
Preferably, described basin there are three, it is connected in series by pipeline head and the tail.
Preferably, described production line has three, pipeline is connected in parallel between the resin column in every production line.
Preferably, there is positive resin packed layer in the resin column.
The advantageous effects of the present invention：
The present invention provides a kind of preparation method of Enzyme catalyzed synthesis glutamine dipeptide, and the present invention utilizes biological enzyme synthesis third
Paddy dipeptides；By adding methanol solution, heating is heated, unnecessary enzyme inactivation in reaction solution, protein denaturation flocculation, through activated carbon
Filtering, the enzyme of flocculation is removed, is adsorbed with resin column plus water removes removing chloride, then glutamine dipeptide crude product is eluted with ammoniacal liquor,
It is brilliant with methanol and elutriation after concentrated, obtain the glutamine dipeptide of high-purity.
The preparation method of Enzyme catalyzed synthesis glutamine dipeptide provided by the invention, synthetic route is simple, and raw material is cheap and easily-available, if
It is standby to require simple, it is few with enzyme amount（0.2%）, the reaction time is short（2-3 hours）, unreacted enzyme is removed, improves the pure of finished product
Degree；It is simple to equipment requirement；High income, post processing is simple, and product purity is high；Cost is low, has high economic value and city
Field competitiveness.
The present invention provides a kind of process system of Enzyme catalyzed synthesis glutamine dipeptide, including three production lines, in each production line
Resin column it is parallel with one another, three production lines can produce simultaneously, can also be produced by technique alternate cycles, three production lines
Resin column absorption, elution, regenerative process cycle alternation are carried out, and the acid tail washings of absorption is used for another group of resin regeneration, the 3rd group
Product, the process system economy, efficient, environmental protection are collected in elution, and tail washings effectively can be recycled fully, and discharge capacity is seldom.Mother liquor
Recycling can has three, and head and the tail are connected, and the mother liquor collected and concentrated in the disposing mother liquor tank in downstream returns to the mother liquor of upstream
In recycling can, the glutamine dipeptide crude product mother solution concentration highest collected in the disposing mother liquor tank being directly connected to resin column can return to
Resin column carries out absorb-elute and obtains the higher glutamine dipeptide solution of concentration, is further refining to obtain object.
Brief description of the drawings
Fig. 1 is the schematic diagram of the present invention.
Embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than limitation the scope of the present invention.
Embodiment 1：
180kg methanol is added in 1000L reactors, and is cooled to 0 DEG C；By 78.6kg thionyl chlorides in 0 DEG C of dropwise addition
Enter in reactor；52.5kgL- alanine is disposably added in reactor by thionyl chloride after being added dropwise completely, insulated and stirred
30min, then 25 DEG C are to slowly warm up to, 1h is stirred, then be warming up to 45 DEG C of stirring 2h；50 DEG C of decompressions evaporate most solvents, obtain
Obtain 90kgL- methyl lactamine salt；
Embodiment 2：
180kg methanol is added in 1000L reactors, and is cooled to 10 DEG C；By 78.6kg thionyl chlorides in 10 DEG C of drops
Add in reactor；Lysine is disposably added in reactor by thionyl chloride after being added dropwise completely, insulated and stirred 30min, then
30 DEG C are to slowly warm up to, stirs 1h, then is warming up to 50 DEG C of stirring 2h；50 DEG C of decompressions evaporate most solvents, obtain 82L- third
Propylhomoserin methyl esters salt；
Embodiment 3：
180kg methanol is added in 1000L reactors, and is cooled to 5 DEG C；By 78.6kg thionyl chlorides in 5 DEG C of dropwise additions
Enter in reactor；Lysine is disposably added in reactor by thionyl chloride after being added dropwise completely, insulated and stirred 30min, then is delayed
It is slow to be warming up to 28 DEG C, 1h is stirred, then be warming up to 48 DEG C of stirring 2h；50 DEG C of decompressions evaporate most solvents, obtain 88kgL- third
Propylhomoserin methyl esters salt；
Embodiment 4：
The synthetic method of ALANINE methyl esters is same as above.By 58g Glus（397mmol）Add in 400mL water, drop
Temperature adjusts pH to 9.0 to 10 DEG C, with ammonia spirit, 1.5ml enzyme liquids is added in reaction system, then by ALANINE methyl esters water
Solution（By 82g ALANINE methyl ester hydrochlorides（590mmol）It is dissolved in 100mL water）It is added dropwise in reaction system, drips simultaneously
Ammonification water, makes pH be maintained at 8.9；Enzyme liquid concentration is measured in course of reaction, enzyme liquid is repeatedly added in timing, when pH is constant, reaction knot
Beam；Add methanol（Reaction solution 20%）60 DEG C of inactivation 30min are heated to, are filtered after adding 5 ‰ activated carbon stirring 20min
Remove biology enzyme flocculate；Reaction solution concentration removes methanol, with positive resin adsorption reaction liquid, after removing chloride is eluted with water, then
Product is parsed with 1% ammoniacal liquor, the part for collecting product is concentrated, and it is 50 DEG C to control outer temperature, and being concentrated into solution has solid to go out
It is existing, carry out being cooled to 30 DEG C, add 300ml methanol aqueous solutions, stirring 30min, is cooled to 5 DEG C of filterings, crystallization 4 after being added dropwise completely
Hour, solid obtains the 60g of glutamine dipeptide crude product one in 50 DEG C of decompression dryings.
60g glutamine dipeptides crude product one is dissolved in 60mL water, add 300mL methanol, slow cooling to 10 DEG C, crystallization 4h,
Glutamine dipeptide crude product two is obtained, is dissolved in water, in 40 DEG C, stirs 30min, 5 ‰ activated carbons stirring 30min is added, filtering, centrifuged
Filter, dries pulverizing obtain 51g glutamine dipeptide sterlings, yield 85%, purity 99.0%.According to high performance liquid chromatography（Chinese Pharmacopoeia
The two annex V D of version in 2010）Measure.
Embodiment 5：
The synthetic method of ALANINE methyl esters is same as above.By 58g Glus（397mmol）Add in 400mL water, drop
Temperature adjusts pH to 8.8 to 10 DEG C, with ammonia spirit, 1.5ml enzyme liquids is added in reaction system, then by ALANINE methyl esters water
Solution（By 82g ALANINE methyl ester hydrochlorides（590mmol）It is dissolved in 100mL water）It is added dropwise in reaction system, drips simultaneously
Ammonification water, makes pH be maintained at 9.0；Enzyme liquid concentration is measured in course of reaction, enzyme liquid is repeatedly added in timing, when pH is constant, reaction knot
Beam；Add methanol（The 20% of reaction solution）70 DEG C of inactivation 30min are heated to, add mistake after 7 ‰ activated carbon stirring 30min
Elimination removes biology enzyme flocculate；Reaction solution concentration removes methanol, with positive resin adsorption reaction liquid, after removing chloride is eluted with water,
Product is parsed with 1% ammoniacal liquor again, the part for collecting product is concentrated, and it is 50 DEG C to control outer temperature, and being concentrated into solution has solid
Occur, carry out being cooled to 35 DEG C, add 300ml methanol aqueous solutions, stirring 30min, is cooled to 5 DEG C of filterings after being added dropwise completely, analyses
Brilliant 4 hours, solid obtains the 60.5g of glutamine dipeptide crude product one in 50 DEG C of decompression dryings.
60.5g glutamine dipeptides crude product one is dissolved in 60.5mL water, adds 300mL methanol, slow cooling is to 8 DEG C, crystallization
4h, glutamine dipeptide crude product two is obtained, is dissolved in water, in 40 DEG C, stir 30min, added 7 ‰ activated carbons stirring 30min, filter, from
The heart filters, and dries pulverizing obtains 51.5g glutamine dipeptide sterlings, yield 86%, purity 99.2%.According to high performance liquid chromatography（China
Two annex V D of pharmacopeia version in 2010）Measure.
Embodiment 6：
The synthetic method of ALANINE methyl esters is same as above.By 58g Glus（397mmol）Add in 400mL water, drop
Temperature adjusts pH to 8.8 to 10 DEG C, with ammonia spirit, 1.5ml enzyme liquids is added in reaction system, then by ALANINE methyl esters water
Solution（By 82g ALANINE methyl ester hydrochlorides（590mmol）It is dissolved in 100mL water）It is added dropwise in reaction system, drips simultaneously
Ammonification water, makes pH be maintained at 9.0；Enzyme liquid concentration is measured in course of reaction, enzyme liquid is repeatedly added in timing, when pH is constant, reaction knot
Beam；Add methanol（The 20% of reaction solution）80 DEG C of inactivation 30min are heated to, add mistake after 7 ‰ activated carbon stirring 30min
Elimination removes biology enzyme flocculate；Reaction solution concentration removes methanol, with positive resin adsorption reaction liquid, after removing chloride is eluted with water,
Product is parsed with 1% ammoniacal liquor again, the part for collecting product is concentrated, and it is 50 DEG C to control outer temperature, and being concentrated into solution has solid
Occur, carry out being cooled to 30 DEG C, add 300ml methanol aqueous solutions, stirring 30min, is cooled to 5 DEG C of filterings after being added dropwise completely, analyses
Brilliant 4 hours, solid obtains the 61.5g of glutamine dipeptide crude product one in 50 DEG C of decompression dryings.
61.5g glutamine dipeptides crude product one is dissolved in 61.5mL water, adds 300mL methanol, slow cooling is to 5 DEG C, crystallization
4h, glutamine dipeptide crude product two is obtained, is dissolved in water, in 40 DEG C, stir 30min, added 7 ‰ activated carbons stirring 30min, filter, from
The heart filters, and dries pulverizing obtains 51.8g glutamine dipeptide sterlings, yield 87%, purity 99.5%.According to high performance liquid chromatography（China
Two annex V D of pharmacopeia version in 2010）Measure.
Embodiment 7：
The synthetic method of ALANINE methyl esters is same as above.By 58g Glus（397mmol）Add in 400mL water, drop
Temperature adjusts pH to 9.2 to 10 DEG C, with ammonia spirit, 1.5ml enzyme liquids is added in reaction system, then by ALANINE methyl esters water
Solution（By 82g ALANINE methyl ester hydrochlorides（590mmol）It is dissolved in 100mL water and is added dropwise in reaction system, drips simultaneously
Ammonification water, makes pH be maintained at 9.0；Enzyme liquid concentration is measured in course of reaction, enzyme liquid is repeatedly added in timing, when pH is constant, reaction knot
Beam；Add methanol（The 20% of reaction solution）80 DEG C of inactivation 30min are heated to, add mistake after 7 ‰ activated carbon stirring 30min
Elimination removes biology enzyme flocculate；Reaction solution concentration removes methanol, with positive resin adsorption reaction liquid, after removing chloride is eluted with water,
Product is parsed with 1% ammoniacal liquor again, the part for collecting product is concentrated, and it is 50 DEG C to control outer temperature, and being concentrated into solution has solid
Occur, carry out being cooled to 40 DEG C, add 300ml methanol aqueous solutions, stirring 30min, is cooled to 8 DEG C of filterings after being added dropwise completely, analyses
Brilliant 4 hours, solid obtains the 59.6g of glutamine dipeptide crude product one in 50 DEG C of decompression dryings.
59.6g glutamine dipeptides crude product one is dissolved in 59mL water, add 300mL methanol, slow cooling to 5 DEG C, crystallization 4h,
Glutamine dipeptide crude product two is obtained, is dissolved in water, in 40 DEG C, stirs 30min, 7 ‰ activated carbons stirring 30min is added, filtering, centrifuged
Filter, dries pulverizing obtain 51.2g glutamine dipeptide sterlings, yield 83%, purity 99.0%.According to high performance liquid chromatography（Chinese Pharmacopoeia
The two annex V D of version in 2010）Measure.
Such as Fig. 1, a kind of process system of Enzyme catalyzed synthesis glutamine dipeptide, including production line, the production line include the ammonia of L- third
Sour 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, mother liquor return
Closed cans 7；Described ALANINE methyl esters synthesis reactor 1 is located at the upstream of described ALANINE methyl esters basin 2, passes through pipeline therebetween
Connection, the upstream of the third described paddy synthesis reaction vessel 3 respectively with the ALANINE methyl esters basin 2, the enzyme basin 4, ammoniacal liquor
Feed pot pipeline is connected, and the downstream of the third paddy synthesis reaction vessel 3 is connected with the charging aperture pipeline of the basin 5；The resin column 5
The discharging opening with the basin 5, acid tube road, ammoniacal liquor pipeline, waterpipe are connected respectively for upstream；Under described resin column 5
Pipeline connects between trip and the 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, every
Pipeline is connected in parallel between resin column 6 in production line；There is positive resin packed layer in the resin column 6.
The synthesis of ALANINE methyl esters is carried out in ALANINE methyl esters synthesis reactor 1, is subsequently collected in ALANINE methyl esters
Basin 2, when starting to synthesize glutamine dipeptide, ALANINE methyl esters basin 2, enzyme basin 4 and ammoniacal liquor pipeline synthesize to the third paddy simultaneously
Reactor 3 feeds, reacted, and reaction solution is collected in basin 5, then heating flocculation, carries out enzyme inactivation, and the solution after filtering enters
Resin column is adsorbed, eluted.
The present invention provides a kind of process system of Enzyme catalyzed synthesis glutamine dipeptide, including three production lines, in each production line
Resin column it is parallel with one another, three production lines can produce simultaneously, can also be produced by technique alternate cycles, three production lines
Resin column absorption, elution, regenerative process cycle alternation are carried out, and the acid tail washings of absorption is used for another group of resin regeneration, the 3rd group
Product, the process system economy, efficient, environmental protection are collected in elution, and tail washings effectively can be recycled fully, and discharge capacity is seldom.Mother liquor
Recycling can has three, and head and the tail are connected, and the mother liquor collected and concentrated in the disposing mother liquor tank in downstream returns to the mother liquor of upstream
In recycling can, the glutamine dipeptide crude product mother solution concentration highest collected in the disposing mother liquor tank being directly connected to resin column can return to
Resin column carries out absorb-elute and obtains the higher glutamine dipeptide solution of concentration, is further refining to obtain object.
Present specification describes some embodiments, it should be understood, however, that those skilled in the art are by reading this specification
It would know that the various improvement without departing substantially from the spirit and scope of the present invention.Therefore, these other embodiments should also be as being included in institute
In attached Claims scope.
SEQ ID NO：1
MKLKATFLFSFFVLYIHVIAQDILTDSAYVREHYTKVEQLIPMRDGVKLFTAIYIPKDTNNKYPFLINR
TPYTVSPYGADQYKKTLGNFPAMMRKGYIFVYQDVRGKWMSEGTFEDVRPQQSKYTSRKDIDESTDTYDTIDWLIKN
IKNNNGKAGVYGISYPGFYSTASLVNAHPALKAVSPQAPVTDWYIGDDFHHGGALFLMDAFRFMSTFGVPRPHPITP
DKGPKGFEFPIKDQYRFYLNAGTVKNLKETYFADSIKFWNDLFAHPDYDQFWKSRLITPHLTNVQPAVMVVGGFFDA
EDAYGAFKTYQSIEQQNKKNNNILVMGPWFHGGWVRGDGSSFGDIQFNQKTSIHYQEELELPFFEYYLKGQGNFNAA
EANIFLSGSNEWKKFSSWPPKETQQRNLYLHPNGKLSFDKVQRTDSWDEYVSDPNNPVPFQAGVWDSRSREYMVDDQ
RFASTRPDVMTYQTDALSEDMTLTGPVIANLVVSTTGTDADYVVKLIDVYPENSPNNKNTMMAGYQMLVRGEILRGK
YRNDFSKPEAFVPNQITKVNYTLPDVGHTFKKGHRIMIQIQNSWFPLADRNPQKFMNIYEAEPQDFQKATQRIFHDV
HNSSYITLPVLNN
Claims (5)
- A kind of 1. preparation method of Enzyme catalyzed synthesis glutamine dipeptide, using following device：Including production line, the production line includes L- Methyl lactamine 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）Positioned at described ALANINE methyl esters basin （2）Upstream, connected therebetween by pipeline, the third described paddy synthesis reaction vessel（3）Upstream respectively with the ALANINE first Ester basin（2）, the enzyme basin（4）, ammoniacal liquor feed pot pipeline connection, the third paddy synthesis reaction vessel（3）Downstream and the storage Tank（5）Charging aperture pipeline connection；The resin column（6 ）Upstream respectively with the basin（5）Discharging opening, acid tube road, ammonia Waterpipe, waterpipe are connected；Described resin column（5）Downstream and the disposing mother liquor tank（7）Between pipeline connect, it is special Sign is, comprises the following steps：（1）The synthesis of ALANINE methyl ester hydrochloride；A. methanol is added in reactor, and dropped Temperature is to 0-10 DEG C；B. thionyl chloride is added dropwise in reactor at 0-10 DEG C；C. by alanine after thionyl chloride is added dropwise completely Disposably it is added in reactor, insulated and stirred 20min-30min, then is to slowly warm up to 25-30 DEG C, stirs 30min-1h, then It is warming up to 45-50 DEG C of stirring 1-2h；D. 50 DEG C of decompressions evaporate most solvents, obtain ALANINE methyl ester hydrochloride；（2） The synthesis of glutamine dipeptide；A. by above-mentioned steps prepare ALANINE methyl ester hydrochloride it is soluble in water and be cooled to 0-10 DEG C it is standby With；B. add water in reactor and be cooled to 5-10 DEG C；C. Glu is added in reactor, and in the temperature PH to 8.8-9.2 is adjusted with ammoniacal liquor under degree, biology enzyme is added in reaction system；The biology enzyme is SEQ ID NO:Shown in 1 Amino acid sequence；The total enzyme amount 50-60% biology enzyme is added for the first time；D. by the ALANINE methyl ester hydrochloride aqueous solution Slowly it is added dropwise at 5-10 DEG C in reactor, adjusts pH pH is maintained 8.9-9.0 with ammoniacal liquor, by above-mentioned remaining total enzyme amount 50-40% Biology enzyme second add in reactor, stirring reaction generation glutamine dipeptide reaction solution；（3）The inactivation of biology enzyme；A. add Methanol solution；B. above-mentioned reaction solution is warming up to 60-80 DEG C；C. filtered after adding 5-7 ‰ activated carbon stirring 20-30min； （4）Filtering；A. above-mentioned reaction solution positive resin acid solution adsorbs；B. resin is eluted with water；C. resin is eluted with ammoniacal liquor, received Integrate cleaning solution as glutamine dipeptide crude product solution；（5）The preparation of glutamine dipeptide crude product one；A. glutamine dipeptide above-mentioned steps obtained Wet product and water are added in dissolution kettle, control 30-35 DEG C of stirring and dissolving of temperature；Ammonification water adjusts pH to 7.3-7.5；B. at 45-50 DEG C Under outer temperature, being concentrated into above-mentioned solution has solid appearance, is cooled to 30-35 DEG C；C. methanol is slowly added dropwise into above-mentioned reactant In system, the mass ratio of methanol and above-mentioned solution is 5-6：1；D. 5-10 DEG C is cooled to after stirring 20-30min after being added dropwise completely, is protected Centrifuged after temperature stirring 1h；E. rinsed to obtain glutamine dipeptide crude product one with the mixed solution of methanol and water；（6）Glutamine dipeptide crude product Two preparation；A. dissolve：The glutamine dipeptide crude product one and the water of 1.0 times of weight that above-mentioned steps are obtained are added 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 into 50 DEG C of outer temperature, being concentrated into solution has Solid occurs, and is cooled to 30-35 DEG C；C. methanol is slowly added dropwise in reaction system；D. after being added dropwise completely after stirring 30min 5-10 DEG C is cooled to, is centrifuged after crystallization insulated and stirred 1h；E. glutamine dipeptide crude product two is rinsed to obtain with the aqueous solution of methanol；（7）Third Paddy dipeptides refines；A. dissolve：The glutamine dipeptide crude product two and water that above-mentioned steps are obtained are added in dissolution kettle, 40 DEG C of stirrings Dissolving；B. decolourize：Reactor adds 5-7 ‰ activated carbon, stirs 30min, filtering；C. above-mentioned filtered fluid is dried, obtained Glutamine dipeptide finished product.
- 2. the preparation method of Enzyme catalyzed synthesis glutamine dipeptide according to claim 1, it is characterised in that in step（2）Third paddy In two peptide symthesis, described biology enzyme, ALANINE methyl ester hydrochloride, ammoniacal liquor are uniformly added into reactor simultaneously；Described biology Enzyme interval is added in reaction system several times.
- 3. the preparation method of Enzyme catalyzed synthesis glutamine dipeptide according to claim 1, it is characterized in that：Described basin（5） There are three, be connected in series by pipeline head and the tail.
- 4. the preparation method of Enzyme catalyzed synthesis glutamine dipeptide according to claim 1, it is characterized in that：Described production line has Three, the resin column in every production line（6）Between pipeline be connected in parallel.
- 5. the preparation method of Enzyme catalyzed synthesis glutamine dipeptide according to claim 1, it is characterized in that：The resin column（6） In have positive resin packed layer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510064439.5A CN104561202B (en) | 2015-02-06 | 2015-02-06 | A kind of preparation method and technology system of Enzyme catalyzed synthesis glutamine dipeptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510064439.5A CN104561202B (en) | 2015-02-06 | 2015-02-06 | A kind of preparation method and technology system of Enzyme catalyzed synthesis glutamine dipeptide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104561202A CN104561202A (en) | 2015-04-29 |
| CN104561202B true CN104561202B (en) | 2018-02-09 |
Family
ID=53078277
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510064439.5A Active CN104561202B (en) | 2015-02-06 | 2015-02-06 | A kind of preparation method and technology system of Enzyme catalyzed synthesis glutamine dipeptide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104561202B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104829680A (en) * | 2015-05-04 | 2015-08-12 | 江苏天晟药业有限公司 | Potassium glycyrrhizate preparation apparatus |
| CN106367340A (en) * | 2016-11-19 | 2017-02-01 | 江苏诚信药业有限公司 | Process system and method for preparing simvastatin |
| 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 |
| CN106699668A (en) * | 2017-02-12 | 2017-05-24 | 江苏诚信药业有限公司 | Process system and method for preparing carnosine |
| 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 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1671840A (en) * | 2002-07-26 | 2005-09-21 | 味之素株式会社 | Novel enzyme forming peptide, microorganism producing the same and process for producing dipeptide using them |
| CN101479292A (en) * | 2006-06-28 | 2009-07-08 | 协和发酵生化株式会社 | Method for purification of oligopeptide |
| CN203284395U (en) * | 2013-05-22 | 2013-11-13 | 新沂市汉菱生物工程有限公司 | Novel device for producing valine |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4447557B2 (en) * | 2003-11-28 | 2010-04-07 | 協和発酵バイオ株式会社 | Dipeptide production method |
| JP4931801B2 (en) * | 2005-03-29 | 2012-05-16 | 協和発酵バイオ株式会社 | Dipeptide crystals and process for producing the same |
| CN101463075A (en) * | 2007-12-19 | 2009-06-24 | 天津天成制药有限公司 | Preparation of N(2)-L-alanyl-L-glutamine |
| CN103087148A (en) * | 2011-11-03 | 2013-05-08 | 青岛康地恩药业股份有限公司 | Synthesis method of N-(2)-L-alanyl-L-glutamine |
| CN102491918B (en) * | 2011-11-28 | 2013-07-10 | 海南灵康制药有限公司 | Alanyl glutamine compound and preparation method thereof |
| CN102838656B (en) * | 2012-09-29 | 2014-10-22 | 德州诚瑞达生物科技有限公司 | Preparation method of L-alanyl-L-glutamine |
| CN103143189A (en) * | 2013-03-12 | 2013-06-12 | 南通诚信氨基酸有限公司 | Process improvement system for preparing alanyl-glutamine |
| CN104163849A (en) * | 2014-08-16 | 2014-11-26 | 厦门世达膜科技有限公司 | Separation and purification method of N(2)-L-alanyl-L-glutamine |
| CN204474686U (en) * | 2015-02-06 | 2015-07-15 | 江苏诚信药业有限公司 | A kind of process system of Enzyme catalyzed synthesis glutamine dipeptide |
-
2015
- 2015-02-06 CN CN201510064439.5A patent/CN104561202B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1671840A (en) * | 2002-07-26 | 2005-09-21 | 味之素株式会社 | Novel enzyme forming peptide, microorganism producing the same and process for producing dipeptide using them |
| CN101479292A (en) * | 2006-06-28 | 2009-07-08 | 协和发酵生化株式会社 | Method for purification of oligopeptide |
| CN203284395U (en) * | 2013-05-22 | 2013-11-13 | 新沂市汉菱生物工程有限公司 | Novel device for producing valine |
Non-Patent Citations (1)
| Title |
|---|
| 二氯亚砜-甲醇溶液中合成氨基酸甲酯盐酸盐的研究;宋霞等;《化学试剂》;20071115;第29卷(第11期);第687页右栏1.3节和第688页左栏第3段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104561202A (en) | 2015-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104561202B (en) | A kind of preparation method and technology system of Enzyme catalyzed synthesis glutamine dipeptide | |
| CN108033893A (en) | The method that continuous flow upstream ultrasonic technique extracts levodopa from cat beans | |
| JPS6124548A (en) | Method for operating ion exchange resin in method for separating basic amino acid | |
| CN102557970B (en) | Preparation method of anhydrous betaine | |
| CN106631853B (en) | A method of extracting levodopa from cat beans | |
| CN101759728A (en) | Method for preparing and refining sucralose | |
| CN102531989B (en) | Purification method for (S)-oxiracetam | |
| CN101407478A (en) | Preparation of creatine hydrochloride | |
| CN101624351A (en) | Preparation method of DL-lysine | |
| CN102286058A (en) | Method for preparing glycyrrhizic acid with high purity | |
| CN109422642B (en) | Refining method of benzoic acid, production method and equipment of pharmaceutical grade sodium benzoate | |
| CN104592053B (en) | A kind of industrialized process for preparing of high-purity sodium pantothenate | |
| CN101985440B (en) | Method for producing piperine | |
| CN111892498A (en) | Method for extracting L-malic acid | |
| CN216192003U (en) | Device system of purification taurine mother liquor | |
| CN106279197A (en) | The purification of isosorbide reaction solution and crystallization processes | |
| CN101811991B (en) | Method for preparing L-arginine nitrate | |
| CN105503630B (en) | A kind of method for purifying lysine hydrochloride | |
| CN107698596A (en) | A kind of synthetic method of Allopurinol | |
| CN101709038A (en) | Method for extracting L-phenylalanine from fermentation broth | |
| CN112662714B (en) | Production process for preparing ornithine | |
| CN1229332C (en) | Extraction process of 15N-L-phenylalanine | |
| CN1065236C (en) | Synthesis of hydrochloride of betaine | |
| CN106755157A (en) | The method for preparing L aspartic acids and L alanine using cis-butenedioic anhydride | |
| CN110483318A (en) | A kind of extracting method of L-threonine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |