CN100488943C - Method for synthesizing theanine - Google Patents

Method for synthesizing theanine Download PDF

Info

Publication number
CN100488943C
CN100488943C CNB200510122360XA CN200510122360A CN100488943C CN 100488943 C CN100488943 C CN 100488943C CN B200510122360X A CNB200510122360X A CN B200510122360XA CN 200510122360 A CN200510122360 A CN 200510122360A CN 100488943 C CN100488943 C CN 100488943C
Authority
CN
China
Prior art keywords
theanine
solution
aminoacylase
weight
acyl
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.)
Expired - Fee Related
Application number
CNB200510122360XA
Other languages
Chinese (zh)
Other versions
CN1789237A (en
Inventor
朱廷哲
张金荣
侯庆林
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tianjin Golden Amino Acid Co ltd
Original Assignee
TIANJIN PHARMACEUTICALS GROUP CORP
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by TIANJIN PHARMACEUTICALS GROUP CORP filed Critical TIANJIN PHARMACEUTICALS GROUP CORP
Priority to CNB200510122360XA priority Critical patent/CN100488943C/en
Publication of CN1789237A publication Critical patent/CN1789237A/en
Application granted granted Critical
Publication of CN100488943C publication Critical patent/CN100488943C/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Landscapes

  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

The present invention relates to a theanine synthesizing method. After L-N-acyl glutamic acid-gamma-ester is ammoniated by a water solution of 70% ethylamine, L-N-acyl theanine is obtained, and is hydThe invention relates to a method for synthesizing theanine, which consists of ammonifying the L-N-acyl aminoglutaric acid-gamma-ester with aminoethane solution of 70% and getting L-N-acyl theanine, hrolyzed by adopting L-aminoacylase, and only the L-N-acyl theanine is hydrolyzed to removed acyl group so as to precipitate L-theanine of high quality out of ethanol water. The present invention combiydrolyzing with L-aminoacylase, and only L-N-acyl theanine is hydrolyzed to remove the acyl, and evoluting in alcohol solution to get the L-theanine of high quality. The invention combines the dominannes the industrialized advantage that the L-aminoacylase hydrolyzes the acyl group, uses the cheap acyl as a protective group, and uses the L-aminoacylase to hydrolyze the acyl group so as to obtain tce of already industrialized L-aminoacylase hydrolyzing acyl, employing cheap acyl as protective group and reacting with aminoethane solution, then hydrolyzing acyl with L-aminoacylase to get L-theanihe L-theanine after reacting with the ethylamine water solution. The present invention has the characteristics of low raw material price, low production cost, high selectivity, high reaction speed andne. The invention is characterized by the cheap raw material, low production cost, high selectivity, fast reaction speed and high productivity. The raw material and agent is suited for industrial prod high yield; the raw material and agents are suitable for industrialized production, and operation process is simple so as to ensure product quality. uction, the operation process is simple and the product quality can be guaranteed.

Description

The synthetic method of theanine
Technical field
The present invention relates to the synthetic method of theanine.
Background technology
Theanine (L-Theanine) is a distinctive amino acid in the tealeaves, accounts for amino acid whosely 50%, and its structural formula is: CH 3CH 2NHCOCH 2CH 2CH (NH 2) COOH.It is a topmost taste compound in the tealeaves.Theanine is a kind of safe, nontoxic, natural additive for foodstuff with different physiological roles.Be widely used in food such as beverage, baked confectionery, frozen dessert etc. abroad, theanine is still blank in the application of field of food at home.Japan has been foodstuff additive in approval L-theanine in 1964, and U.S. FDA was confirmed as the generally recognized as safe material in 1985 with L-theanine.Nearest animal and human's body test shows (Ling Guanting, L-theanine and physiological function thereof, grain and grease.2003.5 46-49): L-theanine is a kind of safety, effectively, excited promotor without any side effects is the good substitute of caffeine.L-theanine is refreshed oneself, and makes resourcefully, focuses one's attention on, and improves learning capacity, brings high blood pressure down, reduces muscle tone pain, separates the fatigue that disappears, regulates the effect of mood, loose spirit.Caffeine-free anxiety waits side effect with feeling sick.Utilize the special physiological function of theanine, develop the functional health medicine, this will play a role to the sub-health state that solves people.
At present the synthetic method of L-theanine mainly comprises the chloroformic acid benzyl ester protection amino (the wine family is more two bright; synthetic [J] Japanese agriculture chemistry of L-theanine can will 1950,23:269), pyrrolidone acid system (CN 02134857), Tetra hydro Phthalic anhydride method (CN 200410014081) wait the amino method of protection to synthesize.Shortcomings such as these synthetic methods exist to some extent that production cost height, speed of response are slow, poor selectivity, yield are low; Adopt aforesaid method, in building-up process, can produce a small amount of D-type product, can not remove, influence quality product with chemical process.
The characteristics of the L-L-Aminoacylase of mentioning among the present invention are the L-type amino acid in can hydrolyzing N-acylamino acid, and this enzyme can extract from animal kidney and obtain; Also can obtain (L-Aminoacylase produces the seed selection of bacterium GR1-11 bacterial strain and the research of condition of enzyme production, Nankai University's journal: natural science edition .1992 (4) .-39-43, Zhou Juyan kingly way guest etc.) by aspergillus or actinomycetes.This enzyme also can be buied from external or domestic reagent market.All can be used for this technology from the L-Aminoacylase that obtains with upper type.At present, domestic many units all adopt different methods to produce this enzyme to be used for the amino acid whose fractionation of DL and to produce L-amino acid (enzyme process of DL-phenylalanine splits research, chemical reaction engineering and technology .2004,20 (1) .-64-69, it is loose that He Shi state Yu one army is permitted literary composition).
N-acyl glutamic acid-γ-ester can make with different methods, and the acyl group in N-acyl glutamic acid-γ-ester is meant and can be preferentially adopted ethanoyl, benzoyl, formyl radical etc. by the acyl group of L-L-Aminoacylase hydrolysis; Ester in N-acyl glutamic acid-γ-ester is meant methyl esters, ethyl ester, butyl ester, benzyl ester etc.; N-acyl glutamic acid-γ-ester can adopt different process, method to produce and obtain.
Summary of the invention
The purpose of this invention is to provide-synthetic method of kind of new theanine, this method has cost of material cheapness, production cost is low, selectivity is high, speed of response is fast, yield is high characteristics, realizes the suitability for industrialized production of this product.
The synthetic method of theanine provided by the invention comprises the steps:
1) R 1OCOCH 2CH 2CH (NH 2) COOH (L-L-glutamic acid-γ-ester) CL is at C 1-4Alcoholic solution or the aqueous solution of alcohol in, add alkali after controlled temperature to 0-40 ℃, add (R 2CO) 2O or R 2To there not being raw material, concentrated after-filtration obtains R to COCl in 0-50 ℃ of reaction 1OCOCH 2CH 2CH (NHCOR 2) COOH (solution of N-acidylate-L-glutamic acid-γ-ester); Or
HOCOCH 2CH 2CH (NHCOR 3) COOH (N-acidylate-L-glutamic acid) adds C 1-6Alcohol, complete molten back react under an acidic catalyst condition and is finished, and adds in the saturated inorganic alkali alcosol and an acidic catalyst makes PH neutrality, filters the concentrated R that obtains in back 1OCOCH 2CH 2CH (NHCOR 3) solution of COOH (N-acidylate-L-glutamic acid-γ-ester);
2) R 1OCOCH 2CH 2CH (NHCOR 3) solution of COOH (N-acidylate-L-glutamic acid-γ-ester) and ethamine or ethylamine solution carry out 0-50 ℃ down reaction obtain N-acidylate-theanine solution;
3) N-acidylate-theanine solution and L-L-Aminoacylase are reacted complete to hydrolysis in 20-60 ℃, filter rear filtrate and concentrate, add C 1-4Alcohol or pure aqueous solution crystallization, obtain the theanine crude product after the filtration;
4) the theanine crude product uses 70-95%C 1-4The aqueous solution of alcohol carry out recrystallization, obtain the theanine elaboration;
R 1=C 1-6Alkyl, R 2=C 2H 5R 3=C 1-8Alkyl.
L-L-glutamic acid-γ-ester: alkali=1:0.1～10 (weight);
L-L-glutamic acid-γ-ester: ethylamine solution=1:1 of ethamine or 50-80%～100; (weight/volume, grams per milliliter);
N-acidylate-L-glutamic acid: ethylamine solution=1:1 of ethamine or 50-80%～100; (weight/volume, grams per milliliter);
L-L-glutamic acid-γ-ester: L-L-Aminoacylase=1:0.1～10 (weight ratio);
N-acidylate-L-glutamic acid: L-L-Aminoacylase=1:0.1～10 (weight ratio);
L-L-Aminoacylase vigor 2000～6000u/g.
Described C 1-4Alcohol be methyl alcohol or ethanol.
Described C 1-6Alcohol be methyl alcohol or ethanol.
Described an acidic catalyst is the vitriol oil.
Described mineral alkali is NaOH, KOH, NaHCO 3, Na 2CO 3
The synthetic method of the theanine of selection provided by the invention comprises the steps:
1) L-L-glutamic acid is dissolved in the dehydrated alcohol, is cooled to 0-10 ℃, adds the vitriol oil, and room temperature reaction 8-10 hour, add alkaline solution to neutral, cooled and filtered, filtration cakes torrefaction obtains the L-ethyl glutamate;
2) the L-ethyl glutamate is dissolved in 30-80% aqueous ethanolic solution, add alkali, the dissolving postcooling is to 0-10 ℃, added the acetic anhydride room temperature reaction 3 hours, concentrate desalination, obtain L-acetylglutamate ethyl ester solution, add 70% ethylamine solution, stirring reaction is complete to aminating reaction, obtains L-acetyl theanine solution;
3) L-acetyl theanine solution is through concentrating, to neutral, temperature 20-60 ℃ adds L-L-Aminoacylase and carries out enzymic hydrolysis, stirring reaction 10 hours is complete to hydrolysis, remove by filter L-L-Aminoacylase, filtrate concentrates, and adds 95% ethanol crystallization, obtain L-theanine crude product, the aqueous ethanolic solution recrystallization of process 80-95% obtains finished product L-theanine;
Wherein, the weight of L-L-glutamic acid and dehydrated alcohol: volume=1:5～15 (grams per milliliter);
The weight of the L-L-glutamic acid and the vitriol oil: volume=1:0.3～2 (grams per milliliter);
L-ethyl glutamate: alkali=1:0.5～3 (weight);
The weight of L-L-glutamic acid and acetic anhydride: volume=1:1～2 (grams per milliliter);
L-ethyl glutamate: 70% ethylamine solution=1:5～15 (weight/volume, grams per milliliter);
L-ethyl glutamate: L-L-Aminoacylase=1:0.5～4 (weight);
L-L-Aminoacylase vigor is 6000u/g;
Described alkali is triethylamine, sodium bicarbonate, NaOH or KOH.
The synthetic method of the theanine of selection provided by the invention comprises the steps:
1) the L-acetylglutamate mixes with methyl alcohol or propyl carbinol, stirs and is cooled to 0-10 ℃, adds the vitriol oil, and room temperature reaction 8-10 hour, be neutralized to neutrality with saturated mineral alkali alcoholic solution, cooled and filtered, mother liquor concentrates, and obtains L-acetylglutamate methyl ester solution;
2) L-acetylglutamate methyl esters or the positive butyl acetate solution of L-acetylglutamate add 70% ethylamine solution, and stirring reaction is complete to aminating reaction, obtain L-acetyl theanine solution;
3) L-acetyl theanine solution is through concentrating, to neutral, temperature 20-60 ℃ adds L-L-Aminoacylase and carries out enzymic hydrolysis, stirring reaction 10 hours is complete to hydrolysis, remove by filter L-L-Aminoacylase, filtrate concentrates, and adds 95% ethanol crystallization, obtain L-theanine crude product, the aqueous ethanolic solution recrystallization of process 80-95% obtains finished product L-theanine;
The weight of L-acetylglutamate and methyl alcohol or propyl carbinol: volume=1:5～15 (grams per milliliter);
The weight of the L-acetylglutamate and the vitriol oil: volume=1:0.3～2 (grams per milliliter);
L-acetylglutamate: alkali=1:0.5～3 (weight);
L-acetylglutamate methyl esters or the positive butyl ester of L-acetylglutamate: 70% ethylamine solution=1:5～15 (weight/volume, grams per milliliter;
L-acetylglutamate: L-L-Aminoacylase=1:0.5～4 (weight);
L-L-Aminoacylase vigor 6000u/g;
Described alkali is triethylamine, sodium bicarbonate, NaOH or KOH.
The synthetic method of the theanine of selection provided by the invention comprises the steps:
1) L-L-glutamic acid is dissolved in the dehydrated alcohol, is cooled to 0-10 ℃, adds the vitriol oil, and room temperature reaction 8-10 hour, add triethylamine solution to neutral, cooled and filtered, filtration cakes torrefaction obtains the L-ethyl glutamate;
2) the L-ethyl glutamate is dissolved in 30-70% aqueous ethanolic solution, add sodium bicarbonate, the dissolving postcooling is to 0-10 ℃, added the acetic anhydride room temperature reaction 3 hours, concentrate desalination, obtain L-acetylglutamate ethyl ester solution, add 70% ethylamine solution, stirring reaction is complete to aminating reaction, obtains L-acetyl theanine solution.
3) L-acetyl theanine solution is through concentrating, to neutral, temperature adds L-L-Aminoacylase for 42 ℃ and carries out enzymic hydrolysis, stirring reaction 10 hours is complete to hydrolysis, remove by filter L-L-Aminoacylase, filtrate concentrates, and adds 95% ethanol crystallization, obtain L-theanine crude product, the aqueous ethanolic solution recrystallization of process 80-95% obtains finished product L-theanine;
Wherein, the weight of L-L-glutamic acid and dehydrated alcohol: volume=1:5～15 (grams per milliliter);
The weight of the L-L-glutamic acid and the vitriol oil: volume=1:0.3～2 (grams per milliliter);
L-ethyl glutamate: alkali=1:0.5～3 (weight);
The weight of L-L-glutamic acid and acetic anhydride: volume=1:1～2 (grams per milliliter);
L-ethyl glutamate: 70% ethylamine solution=1:5～15 (weight/volume, grams per milliliter);
L-ethyl glutamate: L-L-Aminoacylase=1:0.5～4 (weight);
L-L-Aminoacylase vigor is 6000u/g.
The synthetic method of the theanine of selection provided by the invention comprises the steps:
1) the L-acetylglutamate mixes with methyl alcohol or propyl carbinol, stirs and is cooled to 0-10 ℃, adds the vitriol oil, and room temperature reaction 8-10 hour, be neutralized to neutrality with saturated KOH or NaOH ethanolic soln, cooled and filtered, mother liquor concentrates, and obtains L-acetylglutamate methyl ester solution;
2) L-acetylglutamate methyl esters or the positive butyl acetate solution of L-acetylglutamate add 70% ethylamine solution, and stirring reaction is complete to aminating reaction, obtain L-acetyl theanine solution;
3) L-acetyl theanine solution is through concentrating; to neutral; temperature adds L-L-Aminoacylase for 42 ℃ and carries out enzymic hydrolysis; stirring reaction 10 hours is complete to hydrolysis; remove by filter L-L-Aminoacylase, filtrate concentrates, and adds 95% ethanol crystallization; obtain L-theanine crude product, the aqueous ethanolic solution recrystallization of process 80-95% obtains finished product L-theanine.
Wherein, the weight of L-acetylglutamate and methyl alcohol or propyl carbinol: volume=1:5～15 (grams per milliliter);
The weight of the L-acetylglutamate and the vitriol oil: volume=1:0.3～2 (grams per milliliter);
L-acetylglutamate: alkali=1:0.5～3 (weight).
L-acetylglutamate methyl esters or the positive butyl ester of L-acetylglutamate: 70% ethylamine solution=1:5～15; (weight/volume, grams per milliliter)
L-acetylglutamate: L-L-Aminoacylase=1:0.5～4 (weight);
L-L-Aminoacylase vigor 6000u/g.
L-N-acyl glutamic acid-γ-ester obtains L-N-acyl group theanine after 70% ethylamine solution ammonification, also contain a small amount of D-N-acyl group theanine.Adopt the hydrolysis of L-L-Aminoacylase, have only the hydrolysis of L-N-acyl group theanine to remove acyl group, separate out in aqueous ethanolic solution, D-N-acyl group theanine still is dissolved in the alcohol solution, thereby obtains high-quality L-theanine.The present invention uses cheap acyl group to do protecting group in conjunction with the advantage of the L-L-Aminoacylase hydrolysis acyl group of industrialization, after reacting with ethylamine solution, with L-L-Aminoacylase hydrolysis acyl group, just obtains L-theanine again.
The invention provides a kind of synthetic method of new theanine, this method has cost of material cheapness, production cost is low, selectivity is high, speed of response is fast, yield is high characteristics.Raw materials used and reagent is fit to suitability for industrialized production, and operating process is simple, can guarantee quality product.
Embodiment
L-L-glutamic acid-γ-ester can be that raw material prepares as people such as Pan Shirong at " study on the synthesis of L-LEU-glutamic acid methyl ester-L-glutamic acid multipolymer " (biomedical engineering magazine according to currently known methods with L-L-glutamic acid, 1997,2, people such as the method for mentioning 101-104) or what dogface are in " the esterification modification foreign study progress of biosynthesizing polyglutamic acid " (fine chemistry industry, 2002,11,636) the method preparation of mentioning in.L-L-glutamic acid-γ-ethyl ester can directly be bought from Shanghai posthumous title discipline chemical industry company limited etc.
N-acidylate-L-glutamic acid can be that raw material prepares as Shao Weiqing at " preparation of N-p-benzoyl-L-L-glutamic acid " (Chinese Journal of Pharmaceuticals 2004 according to currently known methods with L-L-glutamic acid; 1; people such as method of 10) mentioning or Li Qiuxian are at " N-acylation reaction of L-glutamic acid " (Changchun Polytechnic Univ.'s journal; 2003; 4,19) the method preparation of mentioning in.
N-acidylate-L-glutamic acid-γ-ester and N-acidylate-theanine do not obtain solid materials in reaction process; but exist with the solution form, so be ethamine or ethylamine solution or L-L-Aminoacylase consumption all just L-L-glutamic acid-γ-ester or N-acidylate-L-glutamic acid carry out relevant proportioning and calculate with starting raw material.
Example 1
Take by weighing in 1000 milliliters of three-necked bottles, add L-L-glutamic acid 60 grams, 600 milliliters of dehydrated alcohols are cooled to 10 ℃ under stirring, and drip 30 milliliters of vitriol oils, after drip finishing, room temperature reaction 8 hours drips triethylamine to neutral, cooled and filtered, filtration cakes torrefaction obtains L-L-glutamic acid-γ-ethyl ester 50 grams, yield 70%.
In 1000 milliliters of three-necked bottles, add L-L-glutamic acid-γ-ethyl ester 40g, add 600 milliliters of aqueous ethanolic solutions, add 60 gram sodium bicarbonates, dissolving postcooling to 10 ℃ drips 40 milliliters of acetic anhydrides, drip complete after, room temperature reaction 3 hours, concentrate desalination, obtain just solution of L-N-, add 300 milliliters of 70% ethylamine solutions, stirring reaction is complete to aminating reaction, obtains L-N-acetyl theanine solution.
L-N-acetyl theanine solution is through concentrating; check that solution is to neutral; L-the immobilized aminoacylase that adds equivalent (weight) (is buied from Tianjin TianAn Medicine Industry Co., Ltd; L-L-Aminoacylase vigor 6000u/g) carries out enzymic hydrolysis; 42 ℃ of stirring reactions 10 hours are complete to hydrolysis; remove by filter L-L-Aminoacylase; filtrate concentrates; 95% ethanol crystallization; obtain L-theanine crude product, the aqueous ethanolic solution recrystallization through 80% obtains finished product L-theanine, output 14g; productive rate 35% (in L-L-glutamic acid-γ-ethyl ester) meets japanese food additive standard.
Example 2
Example 1 takes by weighing in 1000 milliliters of three-necked bottles, add L-L-glutamic acid 60 grams, 500 milliliters of propyl carbinols are cooled to below 10 ℃ under stirring, and drip 30 milliliters of vitriol oils, after drip finishing, room temperature reaction 12 hours drips triethylamine to neutral, cooled and filtered, filtration cakes torrefaction obtains L-L-glutamic acid-γ-positive butyl ester 66.6 grams, yield 80%.
In 1000 milliliters of three-necked bottles, add L-L-glutamic acid-γ-positive butyl ester 40g, add 400 milliliters of aqueous ethanolic solutions, add 50 gram sodium bicarbonates, below the dissolving postcooling to 10 ℃, 0 milliliter of acetic anhydride of Dropwise 5, drip finish after, room temperature reaction 2 hours, concentrate desalination, obtain L-N-acetylglutamate-γ-positive butyl acetate solution, add 400 milliliters of 70% ethylamine solutions, stirring reaction is complete to aminating reaction, obtains L-N-acetyl theanine solution.
L-N-acetyl theanine solution is through concentrating; check that solution is to neutral; L-the immobilized aminoacylase that adds equivalent (weight) (is buied from Tianjin TianAn Medicine Industry Co., Ltd; L-L-Aminoacylase vigor 6000u/g) carries out enzymic hydrolysis; 42 ℃ of stirring reactions 10 hours are complete to hydrolysis; remove by filter L-L-Aminoacylase; filtrate concentrates; 95% ethanol crystallization; obtain L-theanine crude product, the aqueous ethanolic solution recrystallization through 80% obtains finished product L-theanine, output 12.35g; productive rate 36% (in L-L-glutamic acid-γ-positive butyl ester) meets japanese food additive standard.
Example 3
In 1000 milliliters of three-necked bottles, add L-L-glutamic acid 60 grams, 600 milliliters of dehydrated alcohols are cooled to 10 ℃ under stirring, drip 30 milliliters of vitriol oils, after drip finishing, room temperature reaction 8 hours drips triethylamine to neutral, cooled and filtered, filtration cakes torrefaction obtain L-L-glutamic acid-γ-ethyl ester 50 grams, yield 70%.
In 1000 milliliters of three-necked bottles, add L-L-glutamic acid-γ-ethyl ester 40g, add 200 milliliters of entry, add 40 gram sodium bicarbonates, dissolving postcooling to 10 ℃ drips 15 milliliters of Benzoyl chlorides, drip complete after, room temperature reaction 2 hours, concentrate desalination, obtain L-N-benzoyl L-glutamic acid-γ-ethyl ester solution, add 400 milliliters of 70% ethylamine solutions, stirring reaction is complete to aminating reaction, obtains L-N-benzoyl theanine solution.
L-N-benzoyl theanine solution is through concentrating; check that solution is to neutral; L-the immobilized aminoacylase that adds equivalent (weight) (is buied from Tianjin TianAn Medicine Industry Co., Ltd; L-L-Aminoacylase vigor 6000u/g) carries out enzymic hydrolysis; 42 ℃ of stirring reactions 16 hours are complete to hydrolysis; remove by filter L-L-Aminoacylase; filtrate concentrates; 95% ethanol crystallization; obtain L-theanine crude product, the aqueous ethanolic solution recrystallization through 80% obtains finished product L-theanine, output 15g; productive rate 37.5% (in L-L-glutamic acid-γ-ethyl ester) meets japanese food additive standard.
Example 4
L-N-acetylglutamate 40g, methyl alcohol 400ml stir to be cooled to and drip the 20ml vitriol oil below 10 ℃, drip and finished the back room temperature reaction 8 hours, be neutralized to neutrality with saturated NaOH alcoholic solution (concentration), cooled and filtered, mother liquor concentrates, and obtains L-N-acetylglutamate-γ-methyl ester solution, adds 300 milliliters of 70% ethylamine solutions, stirring reaction is complete to aminating reaction, obtains L-N-acetyl theanine solution.
L-N-acetyl theanine solution is near neutral through concentrating; 42 ℃ add equivalent immobilized aminoacylases, stirring reaction 10 hours to hydrolysis fully, remove by filter L-Aminoacylase, filtrate concentrates, 95% ethanol is separated out crystallization obtains L-theanine crude product; aqueous ethanolic solution recrystallization through 80% obtains finished product L-theanine 12 grams; yield 32.6% (in the L-N-acetylglutamate) meets japanese food additive standard.
Example 5
L-N-acetylglutamate 40g, 200 milliliters of stirrings of propyl carbinol are cooled to and drip the 20ml vitriol oil below 10 ℃, drip and finished the back room temperature reaction 12 hours, be neutralized to neutrality with saturated KOH alcoholic solution (concentration), cooled and filtered, mother liquor concentrates, and obtains L-N-acetylglutamate-γ-positive butyl acetate solution, adds 300 milliliters of 70% ethylamine solutions, stirring reaction is complete to aminating reaction, obtains L-N-acetyl theanine solution.
L-N-acetyl theanine solution is near neutral through concentrating; 42 ℃ add equivalent immobilized aminoacylases, stirring reaction 10 hours to hydrolysis fully, remove by filter L-Aminoacylase, filtrate concentrates, 95% ethanol is separated out crystallization obtains L-theanine crude product; aqueous ethanolic solution recrystallization through 80% obtains finished product L-theanine 13.3 grams; yield 36.1% (in the L-N-acetylglutamate) meets japanese food additive standard.

Claims (6)

1, a kind of synthetic method of theanine is characterized in that it comprises the steps:
1) R 1OCOCH 2CH 2CH (NH 2) the COOH CL is at C 1-4Alcoholic solution or the aqueous solution of alcohol in, add alkali after controlled temperature to 0-40 ℃, add (R 2CO) 2O or R 2To there not being raw material, concentrated after-filtration obtains R to COCl in 0-50 ℃ of reaction 1OCOCH 2CH 2CH (NHCOR 2) COOH;
2) R 1OCOCH 2CH 2CH (NHCOR 2) solution of COOH and ethamine or ethylamine solution carry out 0-50 ℃ down reaction obtain N-acidylate-theanine solution;
3) N-acidylate-theanine solution and L-L-Aminoacylase are reacted complete to hydrolysis in 20-60 ℃, filter rear filtrate and concentrate, add C 1-4The aqueous solution crystallization of alcohol, obtain the theanine crude product after the filtration;
4) the theanine crude product uses 70-95%C 1-4The aqueous solution of alcohol carry out recrystallization, obtain the theanine elaboration;
R 1=C 1-6Alkyl, R 2=C 2H 5
2, according to the synthetic method of the described theanine of claim 1, it is characterized in that:
R 1OCOCH 2CH 2CH (NH 2) COOH: alkali=1:0.1～10, weight;
R 1OCOCH 2CH 2CH (NH 2) ethylamine solution=1:1～100 of COOH:50-80%; Weight/volume, grams per milliliter;
R 1OCOCH 2CH 2CH (NH 2) COOH:L-L-Aminoacylase=1:0.1～10, weight ratio;
R 1=C 1-6Alkyl;
L-L-Aminoacylase vigor 2000～6000u/g.
3. according to the synthetic method of the described theanine of claim 1, it is characterized in that:
R 1OCOCH 2CH 2CH (NH 2) COOH: alkali=1:0.1～10, weight;
R 1OCOCH 2CH 2CH (NH 2) COOH: ethamine=1:1～100; Weight/volume, grams per milliliter;
R 1OCOCH 2CH 2CH (NH 2) COOH:L-L-Aminoacylase=1:0.1～10, weight ratio;
R 1=C 1-6Alkyl;
L-L-Aminoacylase vigor 2000～6000u/g.
4, according to the synthetic method of the described theanine of claim 1, it is characterized in that: described C 1-4Alcohol be ethanol or methyl alcohol.
5, a kind of synthetic method of theanine is characterized in that it is through following step:
1) L-L-glutamic acid is dissolved in the dehydrated alcohol, is cooled to 0-10 ℃, adds the vitriol oil, and room temperature reaction 8-10 hour, add alkaline solution to neutral, cooled and filtered, filtration cakes torrefaction obtains the L-ethyl glutamate;
2) the L-ethyl glutamate is dissolved in 30-80% aqueous ethanolic solution, add alkali, the dissolving postcooling is to 0-10 ℃, added the acetic anhydride room temperature reaction 3 hours, concentrate desalination, obtain L-acetylglutamate ethyl ester solution, add 70% ethylamine solution, stirring reaction is complete to aminating reaction, obtains L-acetyl theanine solution;
3) L-acetyl theanine solution is through concentrating, to neutral, temperature 20-60 ℃ adds L-L-Aminoacylase and carries out enzymic hydrolysis, stirring reaction 10 hours is complete to hydrolysis, remove by filter L-L-Aminoacylase, filtrate concentrates, and adds 95% ethanol crystallization, obtain L-theanine crude product, the aqueous ethanolic solution recrystallization of process 80-95% obtains finished product L-theanine;
Wherein, the weight of L-L-glutamic acid and dehydrated alcohol: volume=1:5～15, grams per milliliter;
The weight of the L-L-glutamic acid and the vitriol oil: volume=1:0.3～2, grams per milliliter;
L-ethyl glutamate: alkali=1:0.5～3, weight;
The weight of L-L-glutamic acid and acetic anhydride: volume=1:1～2, grams per milliliter;
L-ethyl glutamate: 70% ethylamine solution=1:5～15, weight/volume, grams per milliliter;
L-ethyl glutamate: L-L-Aminoacylase=1:: 0.5～4, weight;
L-L-Aminoacylase vigor is 6000u/g;
Described alkali is triethylamine, sodium bicarbonate, NaOH or KOH.
6, a kind of synthetic method of theanine is characterized in that it is through following step:
1) L-L-glutamic acid is dissolved in the dehydrated alcohol, is cooled to 0-10 ℃, adds the vitriol oil, and room temperature reaction 8-10 hour, add triethylamine solution to neutral, cooled and filtered, filtration cakes torrefaction obtains the L-ethyl glutamate;
2) the L-ethyl glutamate is dissolved in 30-70% aqueous ethanolic solution, add sodium bicarbonate, the dissolving postcooling is to 0-10 ℃, added the acetic anhydride room temperature reaction 3 hours, concentrate desalination, obtain L-acetylglutamate ethyl ester solution, add 70% ethylamine solution, stirring reaction is complete to aminating reaction, obtains L-acetyl theanine solution.
3) L-acetyl theanine solution is through concentrating, to neutral, temperature adds L-L-Aminoacylase for 42 ℃ and carries out enzymic hydrolysis, stirring reaction 10 hours is complete to hydrolysis, remove by filter L-L-Aminoacylase, filtrate concentrates, and adds 95% ethanol crystallization, obtain L-theanine crude product, the aqueous ethanolic solution recrystallization of process 80-95% obtains finished product L-theanine;
Wherein, the weight of L-L-glutamic acid and dehydrated alcohol: volume=1:5～15, grams per milliliter;
The weight of the L-L-glutamic acid and the vitriol oil: volume=1:0.3～2, grams per milliliter;
L-ethyl glutamate: alkali=1:0.5～3, weight;
The weight of L-L-glutamic acid and acetic anhydride: volume=1:1～2, grams per milliliter;
L-ethyl glutamate: 70% ethylamine solution=1:5～15, weight/volume, grams per milliliter;
L-ethyl glutamate: L-L-Aminoacylase=1:0.5～4, weight;
L-L-Aminoacylase vigor is 6000u/g.
CNB200510122360XA 2005-12-15 2005-12-15 Method for synthesizing theanine Expired - Fee Related CN100488943C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CNB200510122360XA CN100488943C (en) 2005-12-15 2005-12-15 Method for synthesizing theanine

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNB200510122360XA CN100488943C (en) 2005-12-15 2005-12-15 Method for synthesizing theanine

Related Child Applications (1)

Application Number Title Priority Date Filing Date
CN2009101369949A Division CN101597239B (en) 2005-12-15 2005-12-15 Method for synthesizing theanine

Publications (2)

Publication Number Publication Date
CN1789237A CN1789237A (en) 2006-06-21
CN100488943C true CN100488943C (en) 2009-05-20

Family

ID=36787387

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB200510122360XA Expired - Fee Related CN100488943C (en) 2005-12-15 2005-12-15 Method for synthesizing theanine

Country Status (1)

Country Link
CN (1) CN100488943C (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101270376B (en) * 2008-05-14 2010-06-02 南京工业大学 Method for synthesizing L-theanine with enzyme
CN102295574B (en) * 2011-08-08 2014-03-26 四川同晟氨基酸有限公司 Method for synthesizing theanine

Also Published As

Publication number Publication date
CN1789237A (en) 2006-06-21

Similar Documents

Publication Publication Date Title
CN101597239B (en) Method for synthesizing theanine
EA019285B1 (en) Process for the stereoselective enzymatic hydrolysis of 5-methyl-3-nitromethyl-hexanoic acid ester
CN102660593A (en) Method for preparing glyceride rich in algal oil n-3 polyunsaturated fatty acid through enzyme process
CN108129288B (en) Synthesis method of trans-3-hydroxycyclobutylformic acid
CN102805290A (en) Method for preparing dietary fiber from wheat bran
CN100488943C (en) Method for synthesizing theanine
CN107986979A (en) A kind of method for synthesizing Beta-alanine calcium and D-VB5 calcium
CN104788332A (en) Preparation and purification method of oleic acid monoethanolamide
CN103319414A (en) Improved telmisartan preparation process
EP2017259A2 (en) Preparation of gamma-amino acids having affinity for the alpha-2-delta protein
CN101563329B (en) Novel method for producing 4,4'-(1-methyl-1,2-ethanediyl)-bis-(2,6-piperazinedione)
CN102686558A (en) Process for production of optically active 3-substituted glutaric acid monoamide
CN103014086A (en) Method for continuously producing L-ornithine composite salt by enzyme immobilization
JPS6188895A (en) Production of stereoisomer of 1-amino-alkyl sulfonic acid or1-amino-alkyl phosphinic acid
CN103044451A (en) Preparation method for cefradine
CN101654694B (en) Preparation method of 3-deacetyl-7-amino-cephemcarboxylic acid
CN109796333B (en) Method for extracting and purifying pravastatin sodium from fermentation liquor
CN102226208B (en) Preparation method of D-asparagine
WO2009087650A2 (en) A novel process for synthesis of pregabalin from substituted cyclopropane intermediate and a process for enzymatic resolution of racemic pregabalin
CN106119307B (en) Preparation method of alpha-ketoglutaric acid
CN102492736B (en) Enzymatic synthesizing method of N-acetyl glycerol glutamate
CN108949844A (en) A kind of enzymatic conversion preparation method of S- methyl-Lcysteine
CN101671704B (en) Method for preparing L-selenomethionine by using enzyme separation method
CN102392064A (en) Production method of bioactive maltose syrup or maltose dextrin
CN102558227A (en) Novel method for preparing high-purity creatine phosphate sodium

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
EE01 Entry into force of recordation of patent licensing contract

Assignee: Tianjin golden amino acid Co.,Ltd.

Assignor: Tianjin Pharmaceuticals Group Corp.

Contract record no.: 2010120000123

Denomination of invention: Synthesis process of theancine

Granted publication date: 20090520

License type: Exclusive License

Open date: 20060621

Record date: 20100913

ASS Succession or assignment of patent right

Owner name: TIANJIN KINGYORK AMINO ACID CO., LTD.

Free format text: FORMER OWNER: TIANJIN PHARMACEUTICALS GROUP CORP.

Effective date: 20121126

C41 Transfer of patent application or patent right or utility model
COR Change of bibliographic data

Free format text: CORRECT: ADDRESS; FROM: 300161 HEDONG, TIANJIN TO: 300457 HANGU, TIANJIN

TR01 Transfer of patent right

Effective date of registration: 20121126

Address after: 300457 No. 223 Huang Hai Road, Tianjin Development Zone

Patentee after: Tianjin golden amino acid Co.,Ltd.

Address before: Cheng linzhuan Hedong District of Tianjin City Road 300161 No. 91

Patentee before: Tianjin Pharmaceuticals Group Corp.

CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20090520

Termination date: 20211215

CF01 Termination of patent right due to non-payment of annual fee