EP1656456A1 - Verfahren zur trennung optischer isomere einer geschützten aminosäure - Google Patents

Verfahren zur trennung optischer isomere einer geschützten aminosäure

Info

Publication number
EP1656456A1
EP1656456A1 EP04748942A EP04748942A EP1656456A1 EP 1656456 A1 EP1656456 A1 EP 1656456A1 EP 04748942 A EP04748942 A EP 04748942A EP 04748942 A EP04748942 A EP 04748942A EP 1656456 A1 EP1656456 A1 EP 1656456A1
Authority
EP
European Patent Office
Prior art keywords
sidechain
carboxyl
protected
chosen
protected aminoacid
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.)
Withdrawn
Application number
EP04748942A
Other languages
English (en)
French (fr)
Inventor
Maxim Ilich Youshko
Vytautas-Juozapas Kajetono Svedas
Roger Arthur Sheldon
Lukas Michael Van Langen
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.)
Clea Technologies BV
Biotir Ltd
Original Assignee
Clea Technologies BV
Biotir Ltd
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 Clea Technologies BV, Biotir Ltd filed Critical Clea Technologies BV
Publication of EP1656456A1 publication Critical patent/EP1656456A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/37Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
    • C07K14/38Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from Aspergillus
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00Preparation of nitrogen-containing organic compounds
    • C12P13/04Alpha- or beta- amino acids
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
    • C12P41/003Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by ester formation, lactone formation or the inverse reactions
    • C12P41/005Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by ester formation, lactone formation or the inverse reactions by esterification of carboxylic acid groups in the enantiomers or the inverse reaction
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
    • C12P41/006Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures
    • C12P41/007Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures by reactions involving acyl derivatives of racemic amines
    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00Technologies relating to chemical industry
    • Y02P20/50Improvements relating to the production of bulk chemicals
    • Y02P20/55Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the present invention relates to a method of enzy- matically separating optical isomers of a protected aminoacid chosen from the group of protected natural and non-natural aminoacids , wherein an enzyme is contacted with a mixture comprising R and S isomers of the protected aminoacid to be separated .
  • Separation of optical isomers is an important procedure in industrial synthesis of aminoacids . Low cost and optical purity (enantiomeric excess ) are important parameters . For this reason , enzymes are used because of their selectivity . To keep cost down, it is generally preferred not to use pure enzyme but rather enzyme preparations .
  • the obj ective of the present invention is to provide a method according to the preambule, allowing the separation at low cost and excellent optical purity ( e . e ) .
  • the present invention is characterized in that the protected aminoacid is a (non-sidechain- carboxyl ) -protected aminoacid having the general formula ( I ) HR 1 N-CR 2 - (CH ) n -C (0) ZR 3 I wherein - R 1 is an amino-protective group or hydrogen; - R 2 is a side chain; - n is a number chosen from 0, 1, and 2; - ZR 3 is a carboxy-protective group where Z is chosen from 0 or NH; and R 1 and R 3 may be integrated, forming a heterocy-oul (non-sidechain-carboxyl) -protected ring, the ring backbone comprising 5 to 8 atoms; the (non-sidechain-carboxyl) -protected
  • the (non-sidechain carboxy) deprotected product carries the now free carboxylgroup.
  • the examples below demonstrate the excellent results obtained with the method according to the invention.
  • the term 'enzymatically separating' is to be understood to increase the number and/or magnitude of properties in which the enzymatically converted product and starting material differ. That is, a physical separation in terms of space is not a requirement, as this may occur later, for example in case of synthesis.
  • a preferred set of substrates having the formula (I) are characterized in that R 3 , with Z is O, represents a group chosen from the set consisting of a branched or unbranched (C ⁇ -C 4 ) alkylgroup, a phenylgroup, and an (C ⁇ -C 4 ) alkylphenylgroup; each of which may be substituted or not with one or more of halogen atoms, car- boxy, hydroxy and aminogroup.
  • Z. is NH
  • R 3 is as defined above, or hydrogen.
  • the nature of the side chain R 2 is believed to be of no particular relevance for the enzymatic reaction.
  • the molecular weight of the (non-sidechain- carboxyl) -protected aminoacid (I) is less than 500.
  • the enzyme is derived from Aspergillus sp., preferably chosen from Aspergillus melleus and Aspergillus oryzae.
  • the (non- sidechain-carboxyl) -protected aminoacid is chosen from a hy- dantoin derivative and diketopiperazine derivative.
  • the enzyme is immobilized, and more preferrably comprised in a CLEA.
  • a CLEA is a cross-linked enzyme aggregate. This pro- vides a cost-effective solution, allowing easy separation of the biocatalytic CLEA and the solution in which the separation is performed.
  • the present invention will now be illustrated with reference to the following examples.
  • use is made of a crude enzyme preparation from Aspergillus, said enzyme preparation having an aminoacylase activity and available from Fluka.
  • This preparation also contains an hydrolase activity exploited in the present invention, more specifically an esterase and/or an amidase activity. Immobilization of enzyme was performed by making a CLEA, according to standard procedures (Cao L. et al, Org. Lett. 2, pp. 1361-1364, (2000) ) .
  • a 10 m solution of D, L-phenylglycine amide (D,L- PGA) was prepared in 5 ml of 50 mM TRIS buffer pH 7.5. Enzymatic reaction was started by addition of 50 U of A inoa- cylase I from Aspergillus melleus (Fluka, 1.3U/mg) and performed at permanent stirring at room temperature. Periodically, samples were taken, diluted by eluent in order to stop enzymatic reaction and subjected to chiral HPLC analysis (Crownpack CR+, pH 2, 25°C) . After 4 hours reaction reached 50.6% conversion, and further hydrolysis was negligible (51% after 18 hours) . Optical purity of remaining D-phenylglycine amide was 99+%, optical purity of converted L-phenylglycine was 99%.
  • D,L-2-aminobutyric acid ethyl ester was prepared by esterification of D, L-2-aminobutyric acid (D,L-ABA, obtained from ACROS) in ethanol in the presence of sulfuric acid. The excess of ethanol was evaporated under reduced pressure. The obtained D,L-ABAE sulfuric salt (24 mmol) was dissolved in 100 ml water and pH was adjusted to 6.7 using NaOH. The reaction was started by adding 1 g of Aminoacylase I from Aspergillus melleus (Fluka, 1.3U/mg) and was performed at the room temperature, at continuous stirring and automated pH-control.
  • Aminoacylase I from Aspergillus melleus

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Mycology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
EP04748942A 2003-06-27 2004-06-25 Verfahren zur trennung optischer isomere einer geschützten aminosäure Withdrawn EP1656456A1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
NL1023767A NL1023767C1 (nl) 2003-06-27 2003-06-27 Werkwijze voor het scheiden van optische isomeren van een beschermd aminozuur.
PCT/RU2004/000244 WO2005001107A1 (en) 2003-06-27 2004-06-25 Method of separating optical isomers of a protected aminoacid

Publications (1)

Publication Number Publication Date
EP1656456A1 true EP1656456A1 (de) 2006-05-17

Family

ID=33550490

Family Applications (1)

Application Number Title Priority Date Filing Date
EP04748942A Withdrawn EP1656456A1 (de) 2003-06-27 2004-06-25 Verfahren zur trennung optischer isomere einer geschützten aminosäure

Country Status (4)

Country Link
EP (1) EP1656456A1 (de)
NL (1) NL1023767C1 (de)
RU (1) RU2006100556A (de)
WO (1) WO2005001107A1 (de)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8934512B2 (en) * 2011-12-08 2015-01-13 Binoptics Corporation Edge-emitting etched-facet lasers

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3647065B2 (ja) * 1994-07-20 2005-05-11 株式会社武蔵野化学研究所 光学活性アラニンの製造方法
JP3160879B2 (ja) * 1996-01-30 2001-04-25 田辺製薬株式会社 光学活性アミノ酸誘導体の製法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2005001107A1 *

Also Published As

Publication number Publication date
RU2006100556A (ru) 2007-08-27
WO2005001107A1 (en) 2005-01-06
NL1023767C1 (nl) 2004-12-28

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