EP0896617A1 - Verfahren zur herstellung von n-geschützten d-prolinderivaten - Google Patents

Verfahren zur herstellung von n-geschützten d-prolinderivaten

Info

Publication number
EP0896617A1
EP0896617A1 EP97914232A EP97914232A EP0896617A1 EP 0896617 A1 EP0896617 A1 EP 0896617A1 EP 97914232 A EP97914232 A EP 97914232A EP 97914232 A EP97914232 A EP 97914232A EP 0896617 A1 EP0896617 A1 EP 0896617A1
Authority
EP
European Patent Office
Prior art keywords
amino acid
acid derivative
proline
protected
cyclic
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Withdrawn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
EP97914232A
Other languages
German (de)
English (en)
French (fr)
Inventor
Martin Sauter
Daniel Venetz
Fabienne Henzen
Diego Schmidhalter
Gabriela Pfaffen
Oleg Werbitzky
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.)
Lonza AG
Original Assignee
Lonza AG
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 Lonza AG filed Critical Lonza AG
Publication of EP0896617A1 publication Critical patent/EP0896617A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
    • C12P41/006—Processes 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
    • C12P41/007—Processes 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/04—Alpha- or beta- amino acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture

Definitions

  • N-protected cyclic D-amino acid derivatives such as B.
  • N-protected D-proline derivatives such as N-benzyloxycarbonyl-D-proline (N-Z-D-proline) are important intermediates for the production of pharmaceuticals (J. Org. Chem., 1994, 59, 7496-7498). So far, only a few enzymes are known which, for. B. Accept N-Z-L-proline as a substrate and hydrolyze it to L-proline.
  • N-acyl-L-proline acylase which, for. B. N-acetyl-L-proline is preferred as the substrate and is used to obtain L-proline.
  • This N-acyl-L-proline acylase is made from microorganisms of the species Comamonas testosteroni or Alcaligenes denitrificans isolated. A disadvantage of these microorganisms is that they are incapable of using NZL-proline as the sole nitrogen source and not hydrolyzing NZL-proline as a substrate.
  • WO 95/10604 describes a microbiological process for producing L-
  • Pipecolic acid known from microorganisms of the species Alcaligenes denitrificans. These microorganisms also have the disadvantage that they do not use the corresponding N-acyl substrate (N-acetyl- (DL) -pipecolic acid) as the only nitrogen source.
  • the object of the present invention is to isolate microorganisms which can be used both for a simple and technically viable process for the preparation of N-protected cyclic or aliphatic D-amino acid derivatives and for a simple process for the preparation of cyclic or aliphatic L-amino acid derivatives. The corresponding products should be isolated in good enantiomeric purity.
  • microorganisms according to claim 1 can be isolated from soil samples, sludge or waste water with the aid of customary microbiological techniques. According to the invention, these microorganisms are isolated in such a way that they are in a medium containing an N-protected proline derivative of the general formula
  • the C 1-4 alkoxy may be applied methoxy, fluorenylmethoxy, ethoxy, propoxy, i-propoxy, butoxy, t-butoxy or i-butoxy.
  • aryl a phenyl or benzyl group is substituted or unsubstituted, such as. B. 4-methoxybenzyl or 4-methoxyphenyl used.
  • Aryloxy is hereinafter defined as a phenyloxy or benzyloxy group, substituted or unsubstituted.
  • Examples of an aryloxy group are benzyloxy, 4-methoxybenzyloxy or 4-nitrobenzyloxy.
  • Sugar alcohol can be used, for example, glycerol.
  • Cultivation and selection are usually carried out at a temperature of 10 to 40 ° C., preferably 20 to 35 ° C. and at a pH between pH 4 and pH 10, preferably between pH 5 and pH 9.
  • Preferred microorganisms are NZL-proline utilizing the genus Arthrobacter (first gram-positive microorganism with proline acylase activity), Agrobacterium / Rhizobium, Bacillus, Pseudomonas or Alcaligenes.
  • microorganisms of the species Arthrobacter sp are NZL-proline utilizing the genus Arthrobacter (first gram-positive microorganism with proline acylase activity), Agrobacterium / Rhizobium, Bacillus, Pseudomonas or Alcaligenes.
  • microorganisms of the species Arthrobacter sp are NZL-proline utilizing the genus Arthrobacter (first gram-positive microorganism with proline acylase activity),
  • HSZ5 with the designation DSM 10328 Agrobacterium / Rhizobium HSZ30, Bacillus simplex K2, Pseudomonas putida K32, Alcaligenes piechaudii K4 or Alcaligenes xylosoxydans ssp denitrificans HSZ 17 with the designation DSM 10329, and their functionally equivalent variants and mutants.
  • the microorganisms DSM 10329 and DSM 10328 were deposited on January 6, 1995 at the German Collection of Microorganisms and Cell Culture GmbH, Mascheroderweg 1b, D-38124 Braunschweig, in accordance with the Budapest Treaty.
  • the partial sequencing of the 16SrDNA showed a similarity of 100% to Bacillus simplex.
  • the profile of cellular fatty acids is typical of Pseudomonas putida.
  • Pseudomonas putida can be assigned.
  • the enzymes according to the invention, the N-acyl-L-proline acylases can, for. B. are obtained by expertly disrupting the microorganism cells described, preferably the enzymes are obtained from Arthrobacter sp HSZ5 (DSM 10329). For example, the ultrasound, French press or lysozyme method can be used for this.
  • the enzymes are characterized by the following properties: N-acyl-L-proline acylase characterized by the following properties: a) substrate specificity:
  • Alkoxy is referred to as a C 1 -, substituted or unsubstituted alkoxy group 18, defined examples of a C 1 - 18 alkoxy group are methoxy, fluorenylmethoxy, ethoxy, propoxy, butoxy, t-butoxy, i-butoxy, Stearoxy.
  • the enriched microorganisms were then separated and cleaned on a solid medium (same composition as liquid medium, only addition of 20 g / l agar agar). In this way, about 30 different bacterial isolates were obtained which were able to use NZL-proline as the only N -Source to recycle.
  • Arthrobacter sp HSZ5 was grown with various C- (NZL-proline as N-source) or N-source (fructose as C-source). C-sources were added to 5 g / l, N-sources to 2 g / l. For induction If necessary, 1 g / l NZL-proline was added to the desired enzymatic activity. Only fructose, glucose, sucrose and mannitol from the tested C sources could be used. In all other cases, N-Z-L-proline was used as the C source. The enzymatic activity was only slightly dependent on the C source used. In contrast, all tested N sources could be used, but the enzymatic activity was, e.g. T. significant, reduced (Table 2):
  • Minimal medium (see example 1) with glucose (30 g / l) and L-proline (7 g / l) as a C or N source at 30 ° C. to a cell density of OD 650 > 35 to induce the enzymatic activity , a small amount of NZ-DL-proline (5 g / l) was then added and incubated for some time. Finally, a further 145 g of NZ-DL-proline were continuously added over a period of 20 hours and then incubated for a further five hours. The cells were then separated by centrifugation.
  • Minimal medium (cf. Example 1) with glucose (20 g / l) and L-proline (7 g / l) as a C or N source at 30 ° C. to a cell density of OD650> 30.
  • 1 12 g of a 50% (w / w) N-Z-DL-proline solution were then added and incubated for a further hour.
  • the volume of the culture was then reduced to 4 l by draining off the amount not required.
  • a further 709 g of the 50% (w / w) N-Z-DL-proline solution were then continuously added to this 4 l culture with induced cells over a period of 5.5 h and then incubated for a further 17.5 h.
  • the pH was maintained at 7.5-8.5 during the biotransformation.
  • Example 1 with glucose (13 g / l) and L-proline (7 g / l) as C or N source at 30 ° C. to the desired cell density (OD650 approx. 25).
  • OD650 approx. 25
  • Figure 2 Activity of the N-acyl-L-proline acylase from Arthrobacter sp. HSZ5 depending on the pH value.
  • Figure 4 Activity of the N-acyl-L-proline acylase from Arthrobacter sp. HSZ5 depending on the concentration of the products.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Analytical Chemistry (AREA)
  • Molecular Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Pyrrole Compounds (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
EP97914232A 1996-03-13 1997-03-12 Verfahren zur herstellung von n-geschützten d-prolinderivaten Withdrawn EP0896617A1 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CH65696 1996-03-13
CH656/96 1996-03-13
PCT/EP1997/001262 WO1997033987A1 (de) 1996-03-13 1997-03-12 Verfahren zur herstellung von n-geschützten d-prolinderivaten

Publications (1)

Publication Number Publication Date
EP0896617A1 true EP0896617A1 (de) 1999-02-17

Family

ID=4192092

Family Applications (1)

Application Number Title Priority Date Filing Date
EP97914232A Withdrawn EP0896617A1 (de) 1996-03-13 1997-03-12 Verfahren zur herstellung von n-geschützten d-prolinderivaten

Country Status (12)

Country Link
US (1) US20020037559A1 (cs)
EP (1) EP0896617A1 (cs)
JP (1) JP2000506728A (cs)
KR (1) KR19990087341A (cs)
CN (1) CN1213400A (cs)
AU (1) AU2155797A (cs)
CA (1) CA2245543A1 (cs)
CZ (1) CZ281198A3 (cs)
NO (1) NO984206L (cs)
PL (1) PL328795A1 (cs)
SK (1) SK282099B6 (cs)
WO (1) WO1997033987A1 (cs)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU5757498A (en) * 1996-12-16 1998-07-15 Lonza A.G. Method for production of d-proline derivatives
JP2002509441A (ja) * 1997-08-11 2002-03-26 ロンザ アーゲー Ｄ−特異的アミノアシラーゼを使用するエナンチオマー的に純粋な環状α−アミノ酸およびそのＮ−保護誘導体の製造方法
DE10050123A1 (de) * 2000-10-11 2002-04-25 Degussa Verfahren zur Herstellung von Aminosäuren
CA2549289A1 (en) * 2003-12-04 2005-06-16 Pfizer Inc. Methods for the preparation of stereoisomerically enriched amines
EP2832345B1 (en) 2012-03-30 2017-11-22 Ajinomoto Co., Inc. Cosmetic composition
CN104592083A (zh) * 2015-01-06 2015-05-06 宁波海硕生物科技有限公司 一种制备n-乙酰-dl-硫代脯氨酸的方法

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4401820A (en) * 1981-01-23 1983-08-30 Tanabe Seiyaku Co., Ltd. Process for racemizing optically active α-amino acids or a salt thereof
US5219741A (en) * 1989-09-06 1993-06-15 Degussa Ag Method of making L-proline using an N-acyl-L-protine acylase
DE3929570A1 (de) * 1989-09-06 1991-03-07 Degussa Mikrobiologisch hergestellte n-acyl-l-prolin-acylase, verfahren zu ihrer gewinnung und ihre verwendung
DE4116980A1 (de) * 1991-05-24 1992-11-26 Degussa Verfahren zur herstellung enantiomerenreiner offenkettiger n-alkyl-l oder d-aminosaeuren

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9733987A1 *

Also Published As

Publication number Publication date
SK282099B6 (sk) 2001-11-06
CZ281198A3 (cs) 1998-12-16
KR19990087341A (ko) 1999-12-27
US20020037559A1 (en) 2002-03-28
CA2245543A1 (en) 1997-09-18
NO984206D0 (no) 1998-09-11
NO984206L (no) 1998-09-11
AU2155797A (en) 1997-10-01
JP2000506728A (ja) 2000-06-06
CN1213400A (zh) 1999-04-07
SK117198A3 (en) 1999-03-12
WO1997033987A1 (de) 1997-09-18
PL328795A1 (en) 1999-02-15

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