CN102392061A - Chemical-enzyme method for preparing D-basic amino acid hydrochloride - Google Patents
Chemical-enzyme method for preparing D-basic amino acid hydrochloride Download PDFInfo
- Publication number
- CN102392061A CN102392061A CN2011102922081A CN201110292208A CN102392061A CN 102392061 A CN102392061 A CN 102392061A CN 2011102922081 A CN2011102922081 A CN 2011102922081A CN 201110292208 A CN201110292208 A CN 201110292208A CN 102392061 A CN102392061 A CN 102392061A
- Authority
- CN
- China
- Prior art keywords
- basic
- amino acid
- hydrochloride
- basic amino
- phenylacetyl
- 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.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a chemical-enzyme method for preparing D-basic amino acid hydrochloride. The method comprises the following steps of: derivatizing DL-basic amino acid serving as a raw material to obtain DL-N-diphenyl acetyl basic amino acid by utilizing an acylating agent, hydrolyzing the DL-N-diphenyl acetyl basic amino acid by using immobilized penicillin acylase as a biocatalyst through non-stereo selectivity and enantioselectivity to obtain L-basic amino acid, phenylacetic acid and D-alpha-N-phenylacetyl basic amino acid; and performing acidolysis on the D-alpha-N-phenylacetyl basic amino acid to obtain phenylacetic acid and D-basic amino acid dihydrochloride, and performing treatment of epoxypropane to obtain the D-basic amino acid hydrochloride. The D-lysine hydrochloride and D-ornithine hydrochloride which are prepared by the method have the yield of more than 40 percent, and ee is more than 99 percent. The method has the characteristics of high yield and optical purity, simple process and the like, and is suitable for the large-scale production of the D-basic amino acid hydrochloride.
Description
Technical field
The present invention relates to a kind of chemo-enzymatic process of the D-of preparation hydrochloride of basic amino acid, belong to biocatalysis and prepare chiral drug midbody technical field.
Background technology
With D-ornithine and D-Methionin is that the D-basic aminoacids of representative is non-protein source chiral amino acid, is prepared as the mono-hydrochloric salts form usually, in chiral drug is synthetic, has extensive use.For example; The verivate of D-ornithine-D-alpha-difluoromethyl ornithine (D-DFMO; According to the good fortune ornithine) be a kind of ornithine decarboxylase irreversible inhibitor, be approved in the U.S. and a kind ofly local be used to remove superfluous hairs in face cream and also possibly be used for oncotherapy.Substitute the tetrapeptide Tyr-D-Orn-Gly-PheNH of second amino acids formation of enkephalin with the D-ornithine
2, its physiological action is stronger tens of times than enkephalin.Mix the D-ornithine in some amino acids microbiotic, strengthen the tolerance of bacterial enzyme degraded, thereby reduce or eliminate the antibiotic resistance of amino acids.D-Methionin is the precursor of synthetic prolan B (LH) analogue, also can be used for synthetic gonadotropin releasing hormone (GnRH) high reactivity analogue.D-Methionin polymer then is the good carrier of medicine.
The preparation method of D-basic aminoacids mainly contains asymmetric synthesis method, chemical resolution method, biological Split Method etc.
1) asymmetric synthesis method.L-Ala and benzaldehyde dimethyl acetal with amido protecting are raw material; Obtain oxazolidone through cyclization; Reacting with γ-iodo tert-butyl acetate, azido-diphenyl phosphate respectively then, is that the via palladium-catalyzed hydrogenation of solvent obtains the D-ornithine derivative at last with methyl alcohol.This method steps is longer, and reagent and expensive raw materials are not suitable for the scale preparation (Bioorg. Med. Lett., 1995,5 (19), 2207) of D-ornithine.
2) chemical resolution method.Liao Xiaoyuan (amino acid and Biological resources, 1999,21 (3), 30) has reported that a kind of is the method that resolving agent prepares D-Methionin with D-tartrate.This method passes through to form D-Methionin-D-tartrate diastereo-isomerism salt, recrystallization, and ion exchange solution salt prepares D-Methionin.The main drawback of this method is that splitting condition is difficult to control, and resolution yield is lower, and D-Methionin optical purity is not high.
3) biological Split Method.DL-ornithine and chloroacetate reaction obtain DL-α, and δ-N-dichloro acetylornithice utilizes the L-Aminoacylase catalytic hydrolysis DL-α in the animal kidney; δ-N-dichloro acetylornithice obtains D-α; δ-dichloro acetylornithice, hydrolysis obtains D-ornithine (J. Biol.Chem., 1951 then; 188 (2), 643).Liu Yi etc. (CN101235403) are raw material with L-l-arginine and hydrochloride thereof, and hydrolysis under alkaline condition, racemization are the DL-ornithine, utilize then hafnia alvei (
Hafnia alvei) L-enantiomorph in the lysine decarboxylase catalysis DL-ornithine among the ASl.1009 is converted into tetramethylenediamine (putrescine), D-ornithine yield is 71～75%.Same bacterial strain also is used for the preparation of D-Methionin.Liu Yi etc. (modern chemical industry, 2007,27 (5), 32) have reported that a kind of chemical-biological coupled method prepares the method for D-Methionin.The racemization of L Methionin chemistry is a DL-Methionin, the employing hafnia alvei (
Hafnia alvei) L-enantiomorph in the lysine decarboxylase catalysis DL-Methionin among the ASl.1009 is converted into pentamethylene diamine, D-Methionin yield is 56.6%.
Summary of the invention
The chemo-enzymatic process that the purpose of this invention is to provide a kind of new preparation D-hydrochloride of basic amino acid.The immobilized penicillin acylated enzyme that utilization has high enantioselectivity splits DL-N-two phenylacetyl basic aminoacidss, obtains L-basic aminoacids, toluylic acid and D-α-N-phenylacetyl basic aminoacids.D-α-N-phenylacetyl basic aminoacids acidolysis is toluylic acid and D-basic aminoacids dihydrochloride, is treated to D-basic aminoacids mono-hydrochloric salts then.Synthetic route is following:
The present invention includes the following step:
(1) DL-two phenylacetyl basic aminoacidss preparation: with the DL-basic aminoacids is raw material and acylating agent stirring reaction 10～20h, and reaction is transferred pH1～2 with hydrochloric acid after accomplishing, and separates out DL-N-two phenylacetyl basic aminoacids solids;
(2) DL-N-two phenylacetyl basic aminoacids enzymatic hydrolysis: under 20～40 ℃ and pH8～10 conditions; 0.1～0.5mol/L DL-two phenylacetyl basic aminoacidss hydrolysis 10～24h under the biological catalyst immobilization penicillin acylated enzyme catalysis; After reaction is accomplished; Concentrated hydrochloric acid is transferred pH1～2, separates out D-α-N-phenylacetyl basic aminoacids;
(3) D-hydrochloride of basic amino acid preparation: D-α-N-phenylacetyl basic aminoacids reaches acidolysis 3～15h under 90～115 ℃ of temperature in 3～8mol/L aqueous hydrochloric acid; Acid hydrolysis solution is used and aqueous hydrochloric acid equal volume of ethyl acetate three times; Evaporated under reduced pressure; The gained solid is dissolved in ethanol or the methyl alcohol, adds propylene oxide, separates out D-basic aminoacids mono-hydrochloric salts.
Raw material DL-basic aminoacids in the step (1) is DL-ornithine or DL-Methionin.
The acylating agent that the DL-basic aminoacids is derived to DL-N-two phenylacetyl basic aminoacidss in the step (1) is toluylic acid, methyl phenylacetate, phenyllacetyl chloride or phenylacetyl bromine; The mol ratio of DL-basic aminoacids and acylating agent is 1:2～6,0～5 ℃ of temperature, pH8～10, reaction times 10～20h.
The concentration of DL-N-two phenylacetyl basic aminoacidss is 0.1～0.5mol/L in the step (2), and the mass ratio of immobilized penicillin acylated enzyme and DL-N-two phenylacetyl basic aminoacidss is 1:2～5; Reaction conditions is: pH8～10,20～40 ℃ of temperature, time 10～4h.
The middle acid hydrolysis solution of step (3) is concentrated into dried, is dissolved in ethanol or the methyl alcohol, adopts propylene oxide that D-basic aminoacids dihydrochloride is converted into D-basic aminoacids mono-hydrochloric salts.
D-hydrochloride of basic amino acid in the step (3) is D-ornithine hydrochloride or D-Methionin mono-hydrochloric salts.
Beneficial effect of the present invention
Compared with prior art; The method that the present invention proposes has the following advantages: 1) adopting the immobilized penicillin acylated enzyme that extensively adopts in the semisynthetic antibiotics industry is biological catalyst; The generation of need not fermenting; Phenylacetyl formation two phenylacetyl basic aminoacidss through two amino of basic aminoacids; Utilize the non-stereoselectivity of immobilized penicillin acylated enzyme and the enantioselectivity of height, obtain D-α-N-phenylacetyl basic aminoacids, guaranteed the high optical activity of D-hydrochloride of basic amino acid; 2) adopt propylene oxide that the dihydrochloride of D-basic aminoacids is converted into mono-hydrochloric salts, simpler than ion exchange separation process, effectively; 3) recycling of circulation fractionation after the racemization of invalid enantiomorph L-basic aminoacids and immobilized penicillin acylated enzyme greatly reduces process cost.
Embodiment
Below be to be biological catalyst with the immobilized penicillin acylated enzyme, prepare the embodiment of D-hydrochloride of basic amino acid through enantioselective hydrolysis, but the present invention be not limited to listed several instances.
Embodiment 1: DL-2, the preparation of 5-N-diphenylacetyl ornithine
52.9g (0.4mol) DL-ornithine and 40g NaOH (1.0mol) are added in the 1000ml water stirring and dissolving.Under condition of ice bath, drip 128ml (0.97mol) phenyllacetyl chloride.After dropwising, reaction is spent the night.Ice bath is cooled to 5 ℃, and suction filtration is used the small amount of deionized water wash solids, gets DL-2,5-N-diphenylacetyl ornithine 135.6g, yield 92% after the drying.
Embodiment 2:DL-2,5-N-diphenylacetyl ornithine enzymatic hydrolysis
With 110.5g (0.3mol) DL-2, the 5-N-diphenylacetyl ornithine adds in the 1000ml water, transfers pH8.0 with ammoniacal liquor.Add the 44g immobilized penicillin acylated enzyme, 30 ℃ of following stirring reaction 24h.Suction filtration is removed immobilized penicillin acylated enzyme.Filtrating is transferred pH1～2 with concentrated hydrochloric acid, suction filtration, solid is used cold water washing, dry D-α-N-phenylacetyl ornithine 31.5g, yield 42%.
Embodiment 3:The preparation of D-ornithine hydrochloride
31.5g D-α-N-phenylacetyl ornithine is dissolved in the 200mL 6 mol/L concentrated hydrochloric acids 90 ℃ of following hydrolysis 12h, ETHYLE ACETATE (200ml * 3) extraction three times; The water layer evaporated under reduced pressure adds 100ml anhydrous alcohol solution solid, adds the 100ml propylene oxide then; Suction filtration, solid washs with small amount of ethanol, oven dry; Get D-ornithine hydrochloride 19.1g, yield 90%, adopting the HPLC method to record ee is 99.6%.
Embodiment 4: DL-2, the preparation of 6-N-hexichol acetyl-l-lysine
58.5g (0.4mol) DL-Methionin and 40g NaOH (1.0mol) are added in the 1000ml water stirring and dissolving.Under condition of ice bath, drip 128ml (0.97mol) phenyllacetyl chloride.After dropwising, reaction is spent the night.Ice bath is cooled to 5 ℃, and suction filtration is used the small amount of deionized water wash solids, gets DL-2,6-N-hexichol acetyl-l-lysine 135g, yield 88% after the drying.
Embodiment 5:DL-2,6-N-hexichol acetyl-l-lysine enzymatic hydrolysis
With 115g (0.3mol) DL-2,6-N-hexichol acetyl-l-lysine adds in the 1000ml water, transfers pH8.0 with ammoniacal liquor.Add the 46g immobilized penicillin acylated enzyme, 30 ℃ of following stirring reaction 24h.Suction filtration is removed immobilized penicillin acylated enzyme.Filtrating is transferred pH1-2 with concentrated hydrochloric acid, suction filtration, solid is used cold water washing, dry D-α-N-phenylacetyl Methionin 32.3g, yield 41%.
Embodiment 6:The preparation of D-lysine hydrochloride
32.3g D-α-N-phenylacetyl Methionin is dissolved in the 200mL 6 mol/L concentrated hydrochloric acids 90 ℃ of following hydrolysis 12h, ETHYLE ACETATE (200ml * 3) extraction three times; The water layer evaporated under reduced pressure adds 100ml anhydrous alcohol solution solid, adds the 100ml propylene oxide then; Suction filtration, solid washs with small amount of ethanol, oven dry; Get D-lysine hydrochloride 19.2g, yield 86%, adopting the HPLC method to record ee is 99.8%.
Claims (6)
1. chemo-enzymatic process for preparing the D-hydrochloride of basic amino acid is characterized in that may further comprise the steps:
(1) DL-two phenylacetyl basic aminoacidss preparation: with the DL-basic aminoacids is raw material and acylating agent stirring reaction 10～20h, and reaction is transferred pH1～2 with hydrochloric acid after accomplishing, and separates out DL-N-two phenylacetyl basic aminoacids solids;
(2) DL-N-two phenylacetyl basic aminoacids enzymatic hydrolysis: under 20～40 ℃ and pH8～10 conditions; 0.1～0.5mol/L DL-N-two phenylacetyl basic aminoacidss hydrolysis 10～24h under the biological catalyst immobilization penicillin acylated enzyme catalysis; After reaction is accomplished; Concentrated hydrochloric acid is transferred pH1～2, separates out D-α-N-phenylacetyl basic aminoacids;
(3) D-hydrochloride of basic amino acid preparation: D-α-N-phenylacetyl basic aminoacids reaches acidolysis 3～15h under 90～115 ℃ of temperature in 3～8mol/L aqueous hydrochloric acid; Acid hydrolysis solution is used and aqueous hydrochloric acid equal volume of ethyl acetate three times; Evaporated under reduced pressure; The gained solid is dissolved in ethanol or the methyl alcohol, adds propylene oxide, separates out the D-hydrochloride of basic amino acid.
2. D-hydrochloride of basic amino acid preparation method according to claim 1 is characterized in that the raw material DL-basic aminoacids in the step (1) is DL-ornithine or DL-Methionin.
3. the preparation method of D-hydrochloride of basic amino acid according to claim 1 is characterized in that the acylating agent that DL-basic aminoacids in the step (1) is derived to DL-N-two phenylacetyl basic aminoacidss is toluylic acid, methyl phenylacetate, phenyllacetyl chloride or phenylacetyl bromine; The mol ratio of DL-basic aminoacids and acylating agent is 1:2～6,0～5 ℃ of temperature, pH8～10, reaction times 10～20h.
4. the preparation method of D-hydrochloride of basic amino acid according to claim 1; The concentration that it is characterized in that DL-N-two phenylacetyl basic aminoacidss in the step (2) is 0.1～0.5mol/L, and the mass ratio of immobilized penicillin acylated enzyme and DL-N-two phenylacetyl basic aminoacidss is 1:2～5; Reaction conditions is: pH8～10,20～40 ℃ of temperature, time 10～24h.
5. the preparation method of D-hydrochloride of basic amino acid according to claim 1; It is characterized in that acid hydrolysis solution is concentrated into dried in the step (3); Be dissolved in ethanol or the methyl alcohol, adopt propylene oxide that D-basic aminoacids dihydrochloride is converted into D-basic aminoacids mono-hydrochloric salts.
6. the preparation method of D-hydrochloride of basic amino acid according to claim 1 is characterized in that the D-hydrochloride of basic amino acid in the step (3) is D-ornithine hydrochloride or D-lysine hydrochloride.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102922081A CN102392061A (en) | 2011-09-29 | 2011-09-29 | Chemical-enzyme method for preparing D-basic amino acid hydrochloride |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102922081A CN102392061A (en) | 2011-09-29 | 2011-09-29 | Chemical-enzyme method for preparing D-basic amino acid hydrochloride |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102392061A true CN102392061A (en) | 2012-03-28 |
Family
ID=45859441
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011102922081A Pending CN102392061A (en) | 2011-09-29 | 2011-09-29 | Chemical-enzyme method for preparing D-basic amino acid hydrochloride |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102392061A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102766060A (en) * | 2012-07-17 | 2012-11-07 | 成都百事兴科技实业有限公司 | Preparation method of D-lysine hydrochloride |
| CN105063117A (en) * | 2015-07-21 | 2015-11-18 | 成都百事兴科技实业有限公司 | Preparation method of D-arginine monohydrochloride |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1526698A (en) * | 2003-03-06 | 2004-09-08 | 四川三高生化股份有限公司 | Production process of high-purity DL-lysine |
| CN101003822A (en) * | 2006-07-05 | 2007-07-25 | 中国科学院成都有机化学有限公司 | Method for producing D amino acid by immobilizing acylation enzyme of penicillin |
| CN101817759A (en) * | 2010-05-12 | 2010-09-01 | 天津天成制药有限公司 | Preparation method of L-carnitine nitrate |
-
2011
- 2011-09-29 CN CN2011102922081A patent/CN102392061A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1526698A (en) * | 2003-03-06 | 2004-09-08 | 四川三高生化股份有限公司 | Production process of high-purity DL-lysine |
| CN101003822A (en) * | 2006-07-05 | 2007-07-25 | 中国科学院成都有机化学有限公司 | Method for producing D amino acid by immobilizing acylation enzyme of penicillin |
| CN101817759A (en) * | 2010-05-12 | 2010-09-01 | 天津天成制药有限公司 | Preparation method of L-carnitine nitrate |
Non-Patent Citations (2)
| Title |
|---|
| 李天童等: "青霉素酰化酶法制备D-丙氨酸", 《精细化工》 * |
| 黄冠华和夏仕文: "酶法拆分D,L-苯丙氨酸制备D-苯丙氨酸", 《合成化学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102766060A (en) * | 2012-07-17 | 2012-11-07 | 成都百事兴科技实业有限公司 | Preparation method of D-lysine hydrochloride |
| CN102766060B (en) * | 2012-07-17 | 2014-04-16 | 成都百事兴科技实业有限公司 | Preparation method of D-lysine hydrochloride |
| CN105063117A (en) * | 2015-07-21 | 2015-11-18 | 成都百事兴科技实业有限公司 | Preparation method of D-arginine monohydrochloride |
| CN105063117B (en) * | 2015-07-21 | 2018-08-17 | 成都百事兴科技实业有限公司 | A kind of preparation method of D-Arg hydrochloride |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JPH09227488A (en) | Production of (s)-cyanohydrin | |
| JP2001037475A (en) | Production of immobilized lipase, enzyme catalytic or enantio-selective conversion of such lipase and substrate, production of optically active compound and use of immobilized lipase | |
| KR20110025917A (en) | Process for the stereoselective enzymatic hydrolysis of 5-methyl-3-nitromethyl-hexanoic acid ester | |
| US6214609B1 (en) | Method and apparatus for preparation of chiral beta amino acids using penicilln G acylase | |
| CN110791538A (en) | Production method suitable for synthesizing sitagliptin phosphate by enzyme method | |
| CN101659622A (en) | Method for splitting valine | |
| JPS592694A (en) | Production of free alpha-amino acid | |
| CN101003822A (en) | Method for producing D amino acid by immobilizing acylation enzyme of penicillin | |
| CN102392061A (en) | Chemical-enzyme method for preparing D-basic amino acid hydrochloride | |
| WO2020192560A1 (en) | Chiral isoquinoline carboxylic acid and preparation method thereof | |
| CN102517351B (en) | A kind of pair of immobilized multienzyme system produces the method for C4H9NO2 | |
| JP4485941B2 (en) | Process for producing optically active α-methylcysteine derivative | |
| CN102352392B (en) | Chemical-enzyme method for preparing D-tyrosine | |
| CN101074446A (en) | Decomposition of tert-leucine with optical activity | |
| WO2004003001A1 (en) | Process for the enzymatic resolution of 1,3-dioxolane-4-carboxylates | |
| CN106480009B (en) | Immobilized amino acid ester acyl transferase and application thereof in preparation of glutamine dipeptide | |
| CN102925530A (en) | L-methionine preparation method | |
| CN101709323A (en) | Method for producing R-mandelic acid with biocatalysis and separating and coupling method | |
| JP2008125364A5 (en) | ||
| JP2003325197A (en) | METHOD FOR ENZYMATIC PRODUCTION OF ENANTIOMERICALLY ENRICHED N-UNPROTECTED beta-AMINO ACID | |
| JP2003325195A (en) | METHOD FOR ENZYMATIC PRODUCTION OF ENANTIOMERICALLY ENRICHED N-UNPROTECTED beta-AMINO ACID, beta-AMINO ACID-n- PROPYL ESTER AND USE THEREOF | |
| CN102226208B (en) | Preparation method of D-asparagine | |
| CN102321695A (en) | Chemical-enzymatic method for preparing D-serine | |
| CN101012181B (en) | Resolution method of DL-p-chlorophenylalanine | |
| CN100391934C (en) | Method for preparing L-glutamic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120328 |