CN101130503B - Method for preparing L-serine-15N - Google Patents

Method for preparing L-serine-15N Download PDF

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CN101130503B
CN101130503B CN2006100302647A CN200610030264A CN101130503B CN 101130503 B CN101130503 B CN 101130503B CN 2006100302647 A CN2006100302647 A CN 2006100302647A CN 200610030264 A CN200610030264 A CN 200610030264A CN 101130503 B CN101130503 B CN 101130503B
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serine
molar weight
acid
reaction
diethyl malonate
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CN101130503A (en
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杨飞
卢伟京
杜晓宁
徐建飞
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Shanghai Research Institute of Chemical Industry SRICI
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Abstract

The invention provides a method for preparing L-serine-<15>N, which comprises the following steps: adopting Na<15>NO2 and diethyl malonate as raw materials to synthesize acetamino-diethyl malonate-<15>N; using the obtained acetamino-diethyl malonate-<15>N to synthesize N-acetyl-DL-serine-<15>N; detaching through enzyme method to obtain the L-serine-<15>N. The utilization rate of the <15>N raw material of the inventive preparation method is high, the optical purity of the purified product L-serine-<15>N is more than 98. 5% and the chemical purity is more than 98%, and the <15>N abundance in the product is more than 98%.

Description

A kind of L-Serine- 15The preparation method of N
Technical field
The present invention relates to preparation method with the organic compound of cold labeling, relate in particular to a kind of L-Serine- 15The preparation method of N.
Background technology
Nitrogen-15 (is called for short 15N, down together) be the stable isotope of nitrogen element, 15The N marker can be used as tracer agent, is widely used in fields such as biological chemistry, medical science, pharmacology, agricultural sciences, especially life science is had very important meaning.Serine- 15N can come postgraduate's object intracellular metabolic as tracer agent.Interpolation L-Serine in the food- 15The N injection can be used to study Serine pathways metabolism (Shemin, D., J.Biol.Chem., 1945,158:297 in animal body; Elwyn, D., Sprinson, D.B., J.Biol.Chem., 1949,178:475), L-serine metabolism relation and proteinic synthesis rate (Monica, C., David in the researching human body, H., Rapid Commun.Mass.Spectrum., 1995,9 (8): 655-659; Stein, T.P., Settle, R.G., Albina, J.A., Denpsey, D.T., Melnick, G., J.Natr.1986,116 (9): 1651～1659), the L-Serine- 15N can be used for preparing other 15N labeled amino acid (Hsiao, H-Y., Wei, T., Campbell, K., Biothechnology and Bioengineering, 1986,28:857～867).By the gene means can with the L-Serine- 15N is used for proteinic selected marker (Waugh, David.S., J.Biomol.NMR., 1996,8 (2): 184～192; Takashi, Y., Takanori, K., Naoshi, D., Jonathan, A., J.Biomol.NMR., 1998,11 (3): 295～306).The L-Serine- 15The application of N more and more widely, good market prospects.
But this type of product production is little, requires high to purity and abundance.Adopt biological method can synthesize the L-Serine- 15N has simple to operate and optical purity of products than characteristics such as height, but 15The N abundance is diluted easily.Prior art have with glycine- 15N is a substrate, the synthetic L-Serine of use hydroxymethyl transferases- 15N (J.P.G.Malthouse., T.B.Fitzpatrick, J.J.Mline, et al, Journal of Peptide Science, 1997,3:361～366); Or with the L-L-Ala- 15N is a substrate, by desaturase prepare the L-Serine- 15N (Kelly, Nicholas.M., O ' Neill, Bridget.C., Probert, John., Reid, Gordon., Stephen, Rosamund., Tetrahedron Lett, 1994,35 (35): 6533～6566).But because two kinds of enzymes being adopted or the enzyme reagent that does not have commodity can be bought in the above-mentioned technology, or the price height is not suitable for being used for producing, and is not suitable method.Use the method for chemistry, have 15Characteristics such as the high and equipment requirements of N abundance is simple, but product is export trade revolve the DL-serine of body- 15N needs to split, just can obtain the L-Serine- 15N.As with the urobenzoic acid ethyl ester- 15N be the synthetic DL-serine of raw material- 15N (Stetten, D., J.Biol.Chem., 1942,144:501), after the fractionation, can obtain the L-Serine- 15N.But this method reactions steps is many, and generated time is long, and yield is not high.As use L-Aminoacylase to split, DL-serine- 15N also need to be converted into N-acetyl-DL-serine- 15N; And N-acetyl-DL-serine- 15The synthetic of N do not appear in the newspapers, usually the yield by the synthetic N-acetyl-DL-serine of DL-serine not high (being about is 31%, S.Akabori, T.T.Otani, R.Marshall, et al, Biochem.Biophys., 1959,83:1).Therefore, need to explore the higher N-acetyl-DL-serine of yield- 15The N route.The method that adopts L-Aminoacylase to split can obtain satisfied optical purity, but do not see be used for N-acetyl-DL-serine- 15The fractionation of N.
Above synthetic method, do not have comparatively suitable preparation L-Serine- 15The N method, need to explore new L-Serine- 15The synthetic route of N, and synthesis technique improved.
Summary of the invention
Purpose of the present invention, provide in order to address the above problem exactly a kind of L-Serine- 15The preparation method of N.
The object of the present invention is achieved like this: a kind of L-Serine- 15The preparation method of N is with Na 15NO 2For the synthetic N-acetyl-DL-serine of isotropic substance nitrogenous source- 15N, with N-acetyl-DL-serine- 15The synthetic L-Serine of N process enzyme fractionation- 15N, concrete steps comprise:
A, with Na 15NO 2With diethyl malonate be raw material, add toluene, water and acetic acid and react, prepare Diethyl Oximinomalonate- 15N;
B, get step a gained Diethyl Oximinomalonate- 15N adds acetate and diacetyl oxide, adopt the reductive method prepare acetamino diethyl malonate- 15N;
C, get step b gained acetamino diethyl malonate- 15N adds formaldehyde and reacts under base catalysis, prepare the methylol acetamino diethyl malonate- 15N;
D, get step c gained methylol acetamino diethyl malonate- 15N adds acid and carries out decarboxylic reaction, prepare N-acetyl-DL-serine- 15N;
E, get steps d gained N-acetyl-DL-serine- 15N adds L-Aminoacylase and splits, through separate purify obtain the L-Serine- 15The N product.
Na described in the step a 15NO 2The mol ratio of reacting with diethyl malonate is 0.5～10: 1, and temperature of reaction is 0～80 ℃, and the reaction times is 3～15 hours, and the adding volume of toluene is 0.5～5 times of raw material diethyl malonate volume, and the add-on of water is raw material Na 15NO 21%～10% of quality, the adding molar weight of acetic acid is raw material Na 15NO 21～10 times of molar weight.
The adding molar weight of the acetate described in the step b be Diethyl Oximinomalonate- 152～10 times of N molar weight, the adding molar weight of diacetyl oxide be Diethyl Oximinomalonate- 151～6 times of N molar weight; Described method of reducing comprises conventional catalytic hydrogenating reduction method and chemical reagent reduction method, and the temperature of reduction reaction is controlled at 0～120 ℃, and reaction pressure is normal pressure～10MPa, and the reaction times is 1～20 hour.
Catalyzer in the described catalytic hydrogenating reduction method is selected from the catalyzer that contains palladium, platinum, ruthenium, rhenium, nickel and composition thereof; chemical reagent in the described chemical reagent reduction method is selected from the particle or the powder of zinc, iron, magnesium, nickel, alumel and composition thereof, the adding molar weight of chemical reagent be Diethyl Oximinomalonate- 151～5 times of N molar weight.
The adding molar weight of the formaldehyde described in the step c be acetamino diethyl malonate- 151～8 times of N molar weight, the adding molar weight of alkali be acetamino diethyl malonate- 151～10 times of N molar weight, temperature of reaction is-10～60 ℃.
Alkali described in the step c is selected from sodium hydroxide, potassium hydroxide, magnesium hydroxide, calcium hydroxide, strontium hydroxide, hydrated barta, yellow soda ash, salt of wormwood and composition thereof.
Acid described in the steps d is selected from formic acid, acetate, propionic acid, butyric acid, phosphoric acid, hydrochloric acid and composition thereof; Acid the adding molar weight be the methylol acetamino diethyl malonate- 152～20 times of N molar weight, the concentration of acid is 1～10mol/L, and temperature of reaction is 30～100 ℃, and the reaction times is 2～6 hours.
L-Aminoacylase described in the step e derives from pluck, aspergillus oryzae, and the condition that splits is concentration of substrate 0.01～1mol/L, enzyme dosage 0.1～4g/L, 30～50 ℃ of temperature, pH value 4～9,8～48 hours time.
Separation described in the step e is purified and is comprised: with the solution concentration behind the resolution reaction, spent ion exchange resin separate obtain the L-Serine- 15The thick product of N, thick product decolours in pH value 3～8 with gac, carries out crystallization at last in the alcoholic acid aqueous solution, obtain the L-Serine- 15The N straight product.
Water and alcoholic acid volume ratio are 1: 0.5～10 in the described alcoholic acid aqueous solution.
Preparation method of the present invention 15N utilization ratio of raw materials height, the product L-Serine that purification obtains- 15The optical purity of N reaches more than 98.5%, and chemical purity reaches more than 98%, in the product 15The N abundance reaches more than 98%.
Embodiment
Below by several embodiment illustrate a kind of L-Serine of the present invention- 15The preparation method of N.
Embodiment 1
With 0.3mol Na 15N0 2(abundance is 99.3%), 0.3mol diethyl malonate, 80ml toluene and 10g water join in the there-necked flask of 500ml and stir, and the temperature of reaction system is reduced to 5 ℃, are added dropwise to 1mol acetate in 1 hour; Temperature is increased to 30 ℃, continues to stir 10 hours.Reaction finishes the back and adds toluene and water, by extracting and separating obtain containing Diethyl Oximinomalonate- 15The toluene phase of N, through washing dry and underpressure distillation remove toluene obtain Diethyl Oximinomalonate- 15N.
With 0.2mol synthetic Diethyl Oximinomalonate- 15N, 0.4mol acetate, 0.8mol diacetyl oxide and 2g 5%Pd/C catalyzer join in the voltage-resistant reactor of 500ml and stir, use nitrogen and hydrogen exchange 10 times respectively, control reaction temperature is 40 ℃, and pressure is 0.1MPa, continues to feed hydrogen reaction 10 hours.Solids removed by filtration, and with the acetate washing leaching cake 4 times of heat, with the acetate pressure reducing and steaming.Add dichloromethane solution with the dissolution of crystals of separating out,, spend the night with anhydrous sodium sulfate drying with saturated common salt water washing 4 times.The methylene dichloride steaming is removed, obtain the crystal of white, with the acetic acid ethyl dissolution of 60ml, add sherwood oil 300ml, placement is spent the night.With the crystal of separating out filter, drying obtains acetamino diethyl malonate- 15N, with the mother liquor secondary crystal can also the recovery part acetamino diethyl malonate- 15N.
Get the 0.1mol acetamino diethyl malonate- 15N, 0.1mol formalin and 20ml water, join in the there-necked flask of 250ml, dripping 10ml concentration is the potassium hydroxide solution of 4mol/L, reacted 3 hours down at 30 ℃, the potassium hydroxide that adds 0.2mol again, with temperature rise to 60 ℃ the reaction 8 hours, obtain the methylol acetamino diethyl malonate- 15The settled solution of N; Add the formic acid of 60ml in solution, temperature rises to 70 ℃, react 4 hours, and underpressure distillation removes anhydrates and acid obtains the melicera material of xanchromatic; Adding acid and water again, to be adjusted to pH be weakly acidic 300ml solution, and last ion exchange resin column washes with water to nearly neutrality, and collection effluent liquid and concentrating under reduced pressure obtain lurid oily matter.The water that adds certain volume obtains the solution of 200ml, adds the 3g gac, 60 ℃ of decolourings 4 hours, filters, and the solution decompression distillation obtains colourless oily mater, and product was placed in the refrigerator 48 hours, obtain N-acetyl-DL-serine- 15The N crystal.
Get 0.1mol N-acetyl-DL-serine- 15It is the 0.5mol/L aqueous solution that N is made into concentration, adds the 0.5g L-Aminoacylase, the pH value be 7 and 40 ℃ of temperature under, carried out enzymatic reaction 36 hours; To split solution concentration for about 200ml, the heating enzymes that went out in 30 minutes are alive down at 100 ℃, and are centrifugal or remove by filter the solid of suspension, the pH value of solution is adjusted to slightly acidic, and last ion exchange resin column washes with water to neutrality, with the ammoniacal liquor wash-out of 1mol/L, collect contain the L-Serine- 15The effluent liquid of N will be collected liquid and concentrate, and obtain the xanchromatic solid; Add 200ml water, add acetate the pH value is adjusted to about 5, add the 1g gac, 50-60 ℃ of decolouring 2 hours, filter, the solution decompression distillation obtains the solid of white; Solid is dissolved in the water of 20ml, adds the ethanol of 30ml under stirring condition, refrigerator is placed and is spent the night, obtain the L-Serine- 15The N crystal.
Embodiment 2
With 1mol Na 15NO 2(abundance is 99.3%), 0.3mol diethyl malonate, 40ml toluene and 4g water join in the there-necked flask of 500ml and stir, and the temperature of reaction system is reduced to 5 ℃, are added dropwise to 5mol acetate in 2 hours; Temperature is increased to 30 ℃, continues to stir 10 hours.Reaction finishes the back and adds toluene and water, by extracting and separating obtain containing Diethyl Oximinomalonate- 15The toluene phase of N, through washing dry and underpressure distillation remove toluene obtain Diethyl Oximinomalonate- 15N.
With 0.2mol synthetic Diethyl Oximinomalonate- 15N, 2mol acetate, 1.2mol diacetyl oxide and 8g5%Pd/C catalyzer join in the voltage-resistant reactor of 500ml and stir, and use nitrogen and hydrogen exchange 10 times respectively, and control reaction temperature is 70 ℃, and pressure is 1.0MPa, continue to feed hydrogen reaction 8 hours.Solids removed by filtration, and with the acetate washing leaching cake 4 times of heat, with the acetate pressure reducing and steaming.Add dichloromethane solution with the dissolution of crystals of separating out,, spend the night with anhydrous sodium sulfate drying with saturated common salt water washing 4 times.The methylene dichloride steaming is removed, obtain the crystal of white, with the acetic acid ethyl dissolution of 60ml, add sherwood oil 300ml, placement is spent the night.With the crystal of separating out filter, drying obtains acetamino diethyl malonate- 15N, with the mother liquor secondary crystal can also the recovery part acetamino diethyl malonate- 15N.
Get the 0.1mol acetamino diethyl malonate- 15N, 0.3mol formalin and 8ml water, join in the there-necked flask of 250ml, Dropwise 5 ml concentration is the sodium hydroxide solution of 4mol/L, reacted 4 hours down at 20 ℃, the sodium hydroxide that adds 0.6mol again, with temperature rise to 40 ℃ the reaction 8 hours, obtain the methylol acetamino diethyl malonate- 15The settled solution of N; Add the acetate of 80ml in solution, temperature rises to 80-90 ℃, react 3 hours, and underpressure distillation removes anhydrates and acid obtains the melicera material of xanchromatic; Adding acid and water again, to be adjusted to pH be weakly acidic 300ml solution, and last ion exchange resin column washes with water to the near neutrality of pH value, and collection effluent liquid and concentrating under reduced pressure obtain lurid oily matter.The water that adds certain volume obtains the solution of 200ml, adds the 2g gac, 80 ℃ of decolourings 2 hours, filters, and the solution decompression distillation obtains colourless oily mater, and product was placed in the refrigerator 48 hours, obtain N-acetyl-DL-serine- 15The N crystal.
Get 0.1mol N-acetyl-DL-serine- 15It is the 1.0mol/L aqueous solution that N is made into concentration, adds the 0.4g L-Aminoacylase, the pH value be 5 and 30 ℃ of temperature under, carried out enzymatic reaction 48 hours; To split solution concentration for about 200ml, the heating enzymes that went out in 50 minutes are alive down at 100 ℃, and are centrifugal or remove by filter the solid of suspension, the pH value of solution is adjusted to slightly acidic, and last ion exchange resin column washes with water to neutrality, with the ammoniacal liquor wash-out of 2mol/L, collect contain the L-Serine- 15The effluent liquid of N will be collected liquid and concentrate, and obtain the xanchromatic solid; Add 200ml water, with acetate solution value is adjusted to slightly acidic, add the 2g gac, 40-50 ℃ of decolouring 4 hours, filter, the solution decompression distillation obtains the solid of white; Solid is dissolved in the water of 20ml, adds the ethanol of 40ml under stirring condition, refrigerator is placed and is spent the night, obtain the L-Serine- 15The N crystal.
Embodiment 3
With 2mol Na 15NO 2(abundance is 99.3%), 0.3mol diethyl malonate, 100ml toluene and 20g water join in the there-necked flask of 500ml and stir, and the temperature of reaction system is reduced to 5 ℃, are added dropwise to 2mol acetate in 4 hours; Temperature is increased to 30 ℃, continues to stir 10 hours.Reaction finishes the back and adds toluene and water, by extracting and separating obtain containing Diethyl Oximinomalonate- 15The toluene phase of N, through washing dry and underpressure distillation remove toluene obtain Diethyl Oximinomalonate- 15N.
With 0.2mol synthetic Diethyl Oximinomalonate- 15N, 1mol acetate, 0.4mol diacetyl oxide join in the voltage-resistant reactor of 500ml and stir, and add excessive zinc powder in batches and carry out reduction reaction, and control reaction temperature is 80 ℃, and pressure is normal pressure, reacts 18 hours.Solids removed by filtration, and with acetate washing leaching cake 3-4 time of heat, with the acetate pressure reducing and steaming.Add dichloromethane solution with the dissolution of crystals of separating out,, spend the night with anhydrous sodium sulfate drying with saturated common salt water washing 4 times.The methylene dichloride steaming is removed, obtain the crystal of white, with the acetic acid ethyl dissolution of 60ml, add sherwood oil 300ml, placement is spent the night.With the crystal of separating out filter, drying obtains acetamino diethyl malonate- 15N, with the mother liquor secondary crystal can also the recovery part acetamino diethyl malonate- 15N.
Get the 0.1mol acetamino diethyl malonate- 15N, 0.6mol formalin and 8ml water, join in the there-necked flask of 250ml, Dropwise 5 ml concentration is the sodium hydroxide solution of 4mol/L, reacted 2 hours down at 40 ℃, the sodium hydroxide that adds 0.8mol again, with temperature rise to 50 ℃ the reaction 8 hours, obtain the methylol acetamino diethyl malonate- 15The settled solution of N; Add the hydrochloric acid of 40ml in solution, temperature rises to 50 ℃, react 6 hours, and underpressure distillation removes anhydrates and acid obtains the melicera material of xanchromatic; Adding acid and water again, to be adjusted to pH be weakly acidic 300ml solution, and last ion exchange resin column washes with water to the near neutrality of pH value, and collection effluent liquid and concentrating under reduced pressure obtain lurid oily matter.The water that adds certain volume obtains the solution of 200ml, adds the 1g gac, 50 ℃ of decolourings 3 hours, filters, and the solution decompression distillation obtains colourless oily mater, and product was placed in the refrigerator 48 hours, obtain N-acetyl-DL-serine- 15The N crystal.
Get 0.1mol N-acetyl-DL-serine- 15It is the 0.05mol/L aqueous solution that N is made into concentration, adds the 4g L-Aminoacylase, the pH value be 5 and 50 ℃ of temperature under, carried out enzymatic reaction 12 hours; To split solution concentration for about 200ml, 100 ℃ down the heating enzyme that went out in 20 minutes live, centrifugal or remove by filter the solid of suspension, with the slightly acidic that is adjusted to of solution, last ion exchange resin column washes with water to neutrality, uses the weak ammonia wash-out, collection contain the L-Serine- 15The effluent liquid of N will be collected liquid and concentrate, and obtain the xanchromatic solid; Add 200ml water, with acetate solution is adjusted to slightly acidic, add the 0.5g gac, 70-80 ℃ of decolouring 1 hour, filter, the solution decompression distillation obtains the solid of white; Solid is dissolved in the water of 20ml, adds the ethanol of 20ml under stirring condition, refrigerator is placed and is spent the night, obtain the L-Serine- 15The N crystal.
To the L-Serine that adopts method of the present invention to prepare in the foregoing description- 15The N product 15The N abundance adopts Delta S gas isotope mass spectrograph (Finnigan company) to carry out analyzing and testing, and adopts Waters2487 high performance liquid chromatography (HPLC) to carry out qualitative and semi-quantitative analysis; Use the optical purity of WZZ-2S polarimeter (Shanghai Precision Scientific Apparatus Co., Ltd) measure product; Adopt the purity of nitriding specimen.Illustrate and adopt preparation method of the present invention, the L-Serine- 15The N product 15The N abundance reaches 98.5%, and optical purity reaches more than 98.5%, and chemical purity reaches 98.6%.

Claims (4)

  1. A L-Serine- 15The preparation method of N is characterized in that, with Na 15NO 2For the synthetic N-acetyl-DL-serine of isotropic substance nitrogenous source- 15N, with N-acetyl-DL-serine- 15The synthetic L-Serine of N process enzyme fractionation- 15N, concrete steps comprise:
    A, with Na 15NO 2With diethyl malonate be raw material, add toluene, water and acetic acid and react, prepare Diethyl Oximinomalonate- 15N;
    B, get step a gained Diethyl Oximinomalonate- 15N adds acetate and diacetyl oxide, adopt the reductive method prepare acetamino diethyl malonate- 15N;
    C, get step b gained acetamino diethyl malonate- 15N adds formaldehyde and reacts under base catalysis, prepare the methylol acetamino diethyl malonate- 15N;
    D, get step c gained methylol acetamino diethyl malonate- 15N adds acid and carries out decarboxylic reaction, prepare N-acetyl-DL-serine- 15N;
    E, get steps d gained N-acetyl-DL-serine- 15N adds L-Aminoacylase and splits, through separate purify obtain the L-Serine- 15The N product;
    Na described in the step a 15NO 2The mol ratio of reacting with diethyl malonate is 0.5～10: 1, and temperature of reaction is 0～80 ℃, and the reaction times is 3～15 hours, and the adding volume of toluene is 0.5～5 times of raw material diethyl malonate volume, and the add-on of water is raw material Na 15NO 21%～10% of quality, the adding molar weight of acetic acid is raw material Na 15NO 21～10 times of molar weight;
    The adding molar weight of the acetate described in the step b be Diethyl Oximinomalonate- 152～10 times of N molar weight, the adding molar weight of diacetyl oxide be Diethyl Oximinomalonate- 151～6 times of N molar weight; Described method of reducing is selected from conventional catalytic hydrogenating reduction method and chemical reagent reduction method, and the temperature of reduction reaction is controlled at 0～120 ℃, and reaction pressure is normal pressure～10MPa, and the reaction times is 1～20 hour;
    The adding molar weight of the formaldehyde described in the step c be acetamino diethyl malonate- 151～8 times of N molar weight, the adding molar weight of alkali be acetamino diethyl malonate- 151～10 times of N molar weight, temperature of reaction is-10～60 ℃;
    Acid described in the steps d is selected from formic acid, acetate, propionic acid, butyric acid, phosphoric acid, hydrochloric acid and composition thereof; Acid the adding molar weight be the methylol acetamino diethyl malonate- 152～20 times of N molar weight, the concentration of acid is 1～10mol/L, and temperature of reaction is 30～100 ℃, and the reaction times is 2～6 hours;
    L-Aminoacylase described in the step e derives from pluck, aspergillus oryzae, and the condition that splits is concentration of substrate 0.01～1mol/L, enzyme dosage 0.1～4g/L, 30～50 ℃ of temperature, pH value 4～9,8～48 hours time;
    Separation described in the step e is purified and is comprised: with the solution concentration behind the resolution reaction, spent ion exchange resin separate obtain the L-Serine- 15The thick product of N, thick product decolours in pH value 3～8 with gac, carries out crystallization at last in the alcoholic acid aqueous solution, obtain the L-Serine- 15The N straight product;
    Chemical reagent in the described chemical reagent reduction method is the particle or the powder of zinc, the adding molar weight of chemical reagent be Diethyl Oximinomalonate- 151～5 times of N molar weight.
  2. 2. L-Serine as claimed in claim 1- 15The preparation method of N is characterized in that: the catalyzer in the described catalytic hydrogenating reduction method is selected from the catalyzer that contains palladium, platinum, ruthenium, rhenium, nickel and composition thereof.
  3. 3. L-Serine as claimed in claim 1- 15The preparation method of N is characterized in that: the alkali described in the step c is selected from sodium hydroxide, potassium hydroxide, magnesium hydroxide, calcium hydroxide, strontium hydroxide, hydrated barta, yellow soda ash, salt of wormwood and composition thereof.
  4. 4. L-Serine as claimed in claim 1- 15The preparation method of N is characterized in that: water and alcoholic acid volume ratio are 1: 0.5～10 in the described alcoholic acid aqueous solution.
CN2006100302647A 2006-08-22 2006-08-22 Method for preparing L-serine-15N Expired - Fee Related CN101130503B (en)

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CN102442921A (en) * 2010-10-13 2012-05-09 中国科学技术大学 Preparation method for 15N/19N dual-labeled unnatural amino acid and intermediates thereof
CN107365260A (en) * 2017-07-31 2017-11-21 长沙道勤生物科技有限公司 The preparation method of feed addictive serine

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2530065A (en) * 1946-12-18 1950-11-14 Sterling Drug Inc Serine synthesis
CN1306092A (en) * 2000-01-18 2001-08-01 覃业亚 Process for preparing levoamino acid by splitting racemic amino acid by immobilized amino acylase
CN1609097A (en) * 2003-10-21 2005-04-27 蔡艳 Serine synthesizing process

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2530065A (en) * 1946-12-18 1950-11-14 Sterling Drug Inc Serine synthesis
CN1306092A (en) * 2000-01-18 2001-08-01 覃业亚 Process for preparing levoamino acid by splitting racemic amino acid by immobilized amino acylase
CN1609097A (en) * 2003-10-21 2005-04-27 蔡艳 Serine synthesizing process

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
H.R.SNYDER ET AL..A CONVENIENT SYNTHESIS OF DL-TRYPTOPHAN.JACS66.1944,350-351. *

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