CZ2001649A3 - Process for preparing (2R)-piperidine derivatives - Google Patents

Abstract

A process for producing (R) -piperidine derivatives of formula (I) and (VII) wherein R 1 is hydroxy, amino or C 1-6 alkoxy and R 4 is hydroxy or amino is described. The (R) -piperidine derivatives are obtained from (RS) -piperidine derivatives of the general formula (II) in which R 2 is hydroxy, amino or C 1-6 alkoxy, in the presence of trace elements, by microorganisms or by cell-free enzymes obtained from these microorganisms

Description

Technical field

The present invention relates to a novel process for the preparation of (2R) -piperidine derivatives using microorganisms or cell-free enzymes derived from these microorganisms.

BACKGROUND OF THE INVENTION

(2R) -piperidine derivatives, such as (R) -pipecolinic acid, are important building blocks of synthesis for biologically active substances such as growth hormone leaching stimulants (WO-A-9513069) or anti-anxiety agents (DE-A- 3702943).

EP-A-686 698 describes a biotechnological process for the preparation of S-.alpha.-aminocarboxylic acids, for example S-.alpha.-pipecolinic acid, by converting racemic amides of RS .alpha.-aminocarboxylic acids with microorganisms of the genera Klebsiella and Pseudomonas. However, in order to obtain pure R-.alpha.-aminocarboxylic acid amides, S-.alpha.-aminocarboxylic acids must be removed by relatively expensive procedures.

There is also a known biotechnological process for producing (R) -pipecolinic acid by culturing microorganisms of the genus Alcaligenes in a medium containing (RS) -pipecolinic acid (Mochizuki, Yamazaki, Y., Maeda, H .: Agric. Biol. Chem., 1988, 52 (5)). , 1113). A drawback of this process is that (R) -pipecolinic acid is obtained in admixture with (S) -2-aminoadipic acid and must be separated after the formation of aminoadipic acid.

(R) -pipecolinic acid is described (J. Org. Chem., 1994, 59, 20975-2081). Here, the esters are converted to the desired ones by isolating them from Aspergillus niger

A process for the production of esters by R.J. Kazlauskas et al. corresponding racemic stereoselective lipases

- 2 00 0 0 0 0 product. The disadvantage of this procedure is that the desired products are of poor enantiomeric purity and are obtained in low yields. In addition, very large amounts of enzyme are also required.

JP-A-63 248 393 discloses a process for producing (R) -pipecolinic acid from racemic pipecolinic acid by microorganisms of the genera Pseudomonas, Kurthia or Alcaligenes. This process has the disadvantage of long reaction times.

It is an object of the present invention to overcome these disadvantages and to provide a simple process for the preparation of (2R) -piperidine derivatives, which can achieve high yields and which can be used in mass production.

This object is solved by a novel biotechnological method according to claim

1.

SUMMARY OF THE INVENTION

The invention relates to a process for the preparation of (2R) -piperidine derivatives of formulas I and VII

wherein R ¹ is hydroxy, amino or C ₁ _ ₆ alkoxy and R ~ ⁴ zanmená hydroxy, or amino.

9

9

9

9

According to the invention, microorganisms are used which have the property of utilizing α-aminocarboxylic acid amides in the form of the racemate or its optically active isomers of formula III as the sole nitrogen source.

9900 99 «9 9900 99 9

(III) wherein A represents together with-NH and -CH optionally substituted 5- or 6-membered saturated heterocyclyl ring. Such microorganisms are known in the art and described in detail in EP-A-686 698.

Preferred microorganisms having the properties described above are microorganisms of the genera Klebsiella and Pseudomonas. Particularly preferably, microorganisms of the Pseudomonas putida species, in particular the Pseudomonas putida species designated DSM 9923, as well as functionally equivalent variants and mutants thereof, are used.

Microorganisms of the species Pseudomonas putida designated DSM 9923 were deposited under the Budapest Convention on 20.04.1995 with the German Collection of Microorganisms and Cell Cultures GmbH (DSM), Mascherodeweg lb, D-38124 Braunschweig.

Functionally equivalent variants and mutants refer to microorganisms having substantially the same properties and functions as the original microorganisms. Such variants and mutants may occur randomly, for example by UV-irradiation.

Surprisingly, these microorganisms or their cell-free enzymes have been found to degrade and, in the case of microorganisms, utilize as the sole source of carbon and energy the S-isomers of the (RS) -piperidine derivatives of formula II

• ·

- 4 wherein R ² represents hydroxy, amino or C ₁ _G-alkoxy, in the presence of trace elements under aerobic conditions at ring opening. After a sufficient reaction time, usually after 1 to 48 hours, there is little or no cyclic .alpha.-aminocarboxylic acids or other interfering open-chain by-products, such as S-2-aminoadipic acid, in the reaction medium. (2R) -piperidine derivatives accumulated in the reaction of formula (I)

IV wherein R ¹ is hydroxy, amino or C ₁ _ ₆ -alkoxy can then be isolated without difficulty, in particular without expensive separation procedures.

C 1 -C 6 -alkoxy means here and in the following, straight or branched chain alkoxy having 1 to 6 carbon atoms. Namely, mention may be made of methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentoxy and its isomers, as well as hexoxy and its isomers.

Enzymes for the cell-free system can be obtained by the known disintegration of microorganisms. For example, ultrasound, French pressure equipment or lysozyme methods may be used. These cell-free enzymes can also be immobilized on a suitable carrier.

The trace elements include, for example, zinc, manganese, cobalt, copper, nickel, molybdenum, calcium, magnesium, boron, iron or sulfur, the optimum composition of the trace elements depending on the microorganism used. Preferably, zinc, manganese, cobalt, copper, nickel, molybdenum, calcium, magnesium, boron, iron and sulfur are used together.

Conveniently, the conversion of the (RS) -piperidine derivatives of the general formula II is effected.

- 5 · «· ······ done with growing microorganisms.

Preferred substrates of formula (II) are (RS) -pipecolinic acid, (RS) -pipecolinic acid ethyl ester, (RS) -pipecolinic acid propyl ester and (RS) -pipecolinic acid amide.

Aerobic conditions are culture conditions in which microorganisms are supplied with oxygen. This can be achieved, for example, by shaking the cultures with air oxygen or by submersing cultures by blowing in molecular oxygen or air oxygen.

The conversion can be effected without conventional cultivation by directly adding the microorganisms to the (RS) -piperidine derivatives of the general formula II. Otherwise, the conversion can take place after the usual cultivation of microorganisms with a suitable source of carbon and energy. As the carbon and energy source, for example, sugars, carboxylic acids, sugar alcohols or amino acids can be used. As sugars, hexoses such as glucose or pentoses can be used. The carboxylic acids which can be used are di- or tricarboxylic acids. salts thereof, such as citric acid or succinate. As the sugar alcohol, a trivalent alcohol such as glycerin can be used. Glutamate can be used as the amino acid.

Preferably, the reaction is carried out without conventional cultivation by directly adding the microorganisms to the (RS) -piperidine derivatives of formula II.

In the reaction, the microorganisms can utilize the (S) -isomer of the (RS) -piperidine derivative of formula (II) as the sole carbon and energy source.

As the medium for the process according to the invention, it is possible to use media customary in the art, such as mineral salt-containing media, for example a mineral salt-containing medium according to Kull et al. (Arch. Microbiol. 135,1-7,1983), media described in Table 1 • 4 • 4 «4 4 4 4

9 9 4 4 5

44 4 4 44 4444

4 or low molar buffers, such as 10 to 100 mM phosphate buffer, to which the necessary trace elements have been added. Preferably, the process is carried out in the media of Table 1.

Suitably, the conditions are selected such that the microorganisms advantageously utilize the S-isomer of the (RS) -piperidine derivatives of formula II as a source of carbon and energy. Thus, preferably the medium contains no other compounds readily usable as a carbon and energy source in addition to compound II.

Suitably, the reaction is carried out with the single, repeated or continuous addition of (RS) -piperidine derivatives of the general formula II. The addition of the (RS) -piperidine derivatives of the general formula II is carried out in such a way that the concentration does not exceed 30% by weight, preferably 10% by weight, particularly preferably 5% by weight.

The pH of the medium is suitably in the range from 4 to 9, preferably from 6 to 8. The reaction is conveniently carried out at a temperature of from 15 to 80 ° C, preferably from 25 to 40 ° C.

The product of formula (I) obtained in the reaction depends on the choice of the starting material and the reaction conditions, in particular the reaction time and the oxygen supply.

Starting from (RS) -pipecolinic acid [(RS) -piperidine II derivative, where R ² is hydroxy], only (2R) -pipecolinic acid can be obtained. Starting from (RS) -pipecolinic acid amide [(RS) -piperidine II derivative, where R ² is amino], both (2R) -pipecolinic acid amide and (2R) -pipecolinic acid can be obtained. When beginning the ester of (RS) -pipecolic [derivative of (RS) -piperidine II wherein R ² is C ₁ _ ₆ alkoxy] may be obtained as an ester (2R) -pipecolic and (2R) -pipecolic .

Whether (2R) -pipecolinic acid amide or (2R) -pipecolinic acid amide is obtained from (RS) -pipecolinic acid amide can be controlled, for example, by reaction time. Thus, when the (RS) -pipecolinic acid amide is converted by the microorganisms of the invention, the (2R) -pipecolinic acid amide, which can optionally be isolated, is first accumulated. After a longer reaction time, the acid is then formed from the (2R) -pipecolinic acid amide. The reaction can be monitored analytically, for example by chromatography. Similarly, in the reaction of (RS) -pipecolinic acid esters, the (2R) -pipecolinic acid ester first accumulates and then the acid.

After a usual reaction time of preferably 1 to 48 hours, (2R) -piperidine derivatives of the formula I, preferably (R) -pipecolinic acid, (R) -pipecolinic acid amide and (R) -pipecolinic acid ethyl and isopropyl ester, are obtained in the medium. in very high to quantitative yield.

The (2R) -piperidine derivatives of formula (I) thus obtained can be isolated by conventional methods such as biomass separation, acidification, chromatography, electrodialysis or crystallization.

Alternatively, the course of the reaction can be controlled by oxygen supply. If, for example, (R) -pipecolinic acid is to be prepared from (RS) -pipecolinic acid amide or (RS) -pipecolinic acid ester, the reaction is expediently carried out under aerobic conditions only after accumulation of the (R) -pipecolinic acid amide, respectively. of the (R) -pipecolinic acid ester as described above, and then the oxygen supply is stopped to form the acid.

Conversion of (RS) -pipecolinic acid derivative of general formula V (V)

wherein R ³ is amino or C ₁ _ _g alkoxy, which C ^ alkoxy has the meaning given above, to the acid (R) -pipecolic IV

(IV) thus takes place via the (R) -pipecolinic acid derivative of the general formula VI

(VI) wherein ^RJ is as defined above, which temporarily accumulates in the medium with increasing enantiomeric excess and which may optionally be isolated. If the (R) -pipecolinic acid derivative of general formula (VI) is to be prepared from the (RS) -pipecolinic acid derivative of general formula (V), it is isolated after accumulation. If (R) -pipecolinic acid of formula (IV) is to be prepared from an (RS) -pipecolinic acid derivative of formula (V), the reaction is expediently carried out after accumulation of the (R) -pipecolinic acid derivative of formula (VI) under aerobic conditions. .

In order to determine whether the (R) -pipecolinic acid derivative of general formula (VI) has accumulated in the medium, the reactions can be monitored analytically, for example by chromatography. The reaction is to be terminated as soon as the maximum accumulation of the (R) -pipecolinic acid derivative of general formula (VI) is reached.

Another aspect of the invention is the further conversion, reduction, of (2R) -pipecolinic acid derivatives of formula (VI) to (2R) -piperidine derivatives of formula (VII).

(VII) wherein R ⁴ is hydroxy or amino.

As the reducing agent, an alkali metal or alkaline earth metal hydride is usually used, such as, for example, aluminum hydride, sodium borohydride, sodium aluminum hydride, aluminum hydride, magnesium borohydride or calcium borohydride. In particular, aluminum hydride is used.

Advantageously, the reaction is carried out at a temperature of from 0 to 100 ° C, preferably at the reflux temperature of the corresponding solvent.

As is known in the art, the reduction can be carried out in a polar organic solvent such as ether. Suitable ethers are, for example, diethyl ether, dipropyl ether, tetrahydrofuran or 1,4-dioxane.

After a usual reaction time of from 1 to 16 hours, the desired products of formula VII, such as (R) -2- (aminoethyl) piperidine or (R) -2- (hydroxymethyl) piperidine, can then be isolated by known processing methods.

Table 1

Medium 1 (NH ₄ ) ₂ Sat ₄ 2,0 g / i Na ₂ HPO ₄ 2,0 g / l kh ₂ Mon ₄ 1.0 g / i NaCl 2,0 g / i MgCl ₂ .6H ₂ O 0.4 g / i CaCl ₂ .2H ₂ 0 14.5 mg / l FeCl ₃ .6H ₂ O 0.8 mg / l ZnSO ₄ .7H ₂ O O, ¹ mg / l MnCl ₂ .4H ₂ O 0.09 mg / l H3BO3 0.3 mg / l CoCl ₂ .6H ₂ 0 0.2 mg / l CuC1 ₂ 2H ₂ O 0.01 mg / l NiCl ₂ .6H ₂ O 0.02 mg / l Na ₂ MoO ₄ .2H ₂ O 0.03 mg / l FeSO ₄ .7H ₂ O 2,0 mg / l EDTA-Na ₂ .2H ₂ 0 5.0 mg / l

0 0 0 «« 00 00 0 • «0 0 · · · · 000 000 00 0 00 0 * 0 0 · 0 0 0« 0 0 0

000 000 000 0000 00 00 0000 00 00 »

Medium 2 kh ₂ after ₄ NH ₄ Cl (NH _{4) 2} SO ₄ Na ₂ SO ₄ .6H ₂ MgCl ₂ O CaCl ₂ .2H ₂ O ZnSO ₄ 7H ₂ O MnCl ₂ .4H ₂ O of H ₃ BO ₃

COC1 ₂ .6H ₂ 0 CuC1 ₂ .2H ₂ O NiCl ₂ .6H ₂ O Na ₂ MO0 ₄ .2H ₂ O FeSO ₄ 7H ₂ O-Na ₂ EDTA 2H ₂ 0

Medium 3

Na ₂ SO ₄

Na ₂ HPO ₄ kh ₂ after ₄

NaCl

MgCl ₂ .6H ₂ O CaCl ₂ .2H ₂ 0 FeCl ₃ .6H ₂ O ZnSO ₄ 7H ₂ O MnCl ₂ .4H ₂ OH ₃ ^b 3 °

CoCl ₂ .6H ₂ O CuCl ₂ .2H ₂ O NIC ₂ .6H ₂ O Na ₂ MoO ₄ .2H ₂ O FeSO ₄ .7H ₂ O EDTA-Na ₂ .2H ₂ 0

1.3 g / l 5.0 g / 1 2,0 g / 1 0.25 g / l 0.8 g / i 0.16 g / l 9.0 mg / l 4.0 mg / l 2.7 mg / l 1,8 mg / l 1.5 mg / l 0.18 mg / L 0.2 mg / l 30.0 mg / l 175, 0 mg / l

o, i g / l 2,0 g / i 1.0 g / 1 2,0 g / i 0.4 g / i 14, E > mg / l 0.8 mg / l 0.1 mg / l 0.09 mg / l 0.3 mg / l 0.2 mg / l 0.01 mg / l 0.02 mg / l 0.03 mg / l 2,0 mg / l 5.0 mg / l

• · · «* * * <

• · a

DETAILED DESCRIPTION OF THE INVENTION

Example 1

Production of (R) -pipecolinic acid from (RS) -pipecolinic acid

1.1 in shaking culture

In a Erlenmeyer flask, Pseudomonas putida DSM 9923 was added to 400 ml of medium 1 and (RS) -pipecolinic acid at a concentration of 1.5%. The contents were incubated at 30 ° C and on a shaker at 140 rpm. After 24 hours, the cell-free supernatant was tested at an OD650 of 3.7 by HPLC for (R) -pipecolinic acid content.

(R) -pipecolinic acid was obtained in a yield of> 45% based on the (RS) -pipecolinic acid introduced and with an ee value of> 99%.

1.2 in a fermenter

Pseudomonas putida DSM 9923 was added to 100 ml of medium 3 and 1% (RS) -pipecolinic acid in a 500 ml bullying flask with bullying. The contents were incubated for 16 hours at 30 ° C and on a shaker at 140 rpm. A pre-culture of Pseudomonas putida DSM 9923 was added to a 51 L fermenter with 3 L medium 2 and (RS) -pipecolinic acid at a concentration of 45% (45 g). After 7 hours, an additional 45 g of acid (RS) were continuously added. -pipecoline as a 10% solution. After a further 4 hours, the biotransformation was terminated by the cell-free supernatant was assayed by HPLC for (R) -pipecolinic acid content. (R) -pipecolinic acid was obtained in quantitative yield and with an ee value of> 97%.

Example 2

Production of (R) -pipecolinic acid from (RS) pipecolinic acid ethyl ester

In a 100 ml Erlenmeyer bullying flask, Pseudomonas putida DSM 9923 was added to 40 ml of medium 3 and 1.5% (RS) -pipecolinic acid ethyl ester. The contents were incubated at 30 ° C on a shaker at 140 rpm. After 26 hours, the cell-free supernatant was tested at OD650 2.0 by HPLC for (R) -pipecolinic acid content. (R) -pipecolinic acid was obtained in a yield of 12 to> 45% based on the (RS) -pipecolinic acid ethyl ester introduced, and with an ee value of> 80%. After 44 hours at OD650 3.2, enantiomerically pure (R) -pipecolinic acid was obtained.

Example 3 • 4 4 · 44

Production of (R) -pipecolinic acid amide from pipecolinic acid amide (RS) in shake culture

In a 100 ml Erlenmeyer bullying flask, add 40 ml of medium and (RS) -pipecolinic acid amide Pseudomonas putida DSM 9923. Shaker contents at 140 rpm. After the supernatant tested at OD650 3.0 at a concentration of 1.5% added was incubated at 30 ° C for 20 hours with cell-free HPLC for the (R) -pipecolinic acid amide content. The (R) -pipecolinic acid amide was obtained in a yield of> 45% based on the (RS) -pipecolinic acid amide introduced and with an ee value of> 99%.

Example 4

Production of (R) -pipecolinic acid amide and (R) -pipecolinic acid from (RS) -pipecolinic acid amide in a Pseudomonas putida DSM 9923 fermenter was grown overnight on medium 3 with 20 g / l glutamate in the fermenter. At an OD 650 of 14, 1% (RS) -pipecolinic acid amide was added. After 2 hours, according to GC analysis, virtually enantiomerically pure (R) -pipecolinic acid amide was present. Pipecolinic acid was not detected.

One sample of this solution was incubated in a shaker at 37 ° C and 140 rpm to close the air inlet. After 48 hours, according to GC analysis, enantiomerically pure (R) -pipecolinic acid was present. Pipecolinic acid amide could no longer be detected.

Example 5

Production of (R) -pipecolinic acid from (RS) -pipecolinic acid amide in shake culture

In a 100 ml Erlenmeyer Bulb with Bullying, add 40 ml of medium

- 13 ♦ · 4 · ·· and (RS) -pipecolinic acid amide Pseudomonas putida DSM 9923. Shaker at 140 rpm. After the supernatant was tested at OD650 3.9 at a concentration of 1% added and incubated at 30 ° C for 26 hours with cell-free GC.

(R) -pipecolinic acid with an ee value of > 98 yield of ca 10% was found.

ve

Example 6

Production of (R) -pipecolinic acid isopropyl ester from (RS) -pipecolinic acid isopropyl ester in shaken culture

In a 500 mL Erlenmeyer Bullying flask, 100 mL of minimal medium 3 was added with 4 g / l (RS) -pipecolinic acid isopropyl ester and Pseudomonas putida DSM 9923. The contents were incubated at 30 ° C on a shaker at 140 rpm. After 4.5 hours, the cell-free supernatant was tested at OD650 of 1.4 with GC for the content and the enantiomeric excess of pipecolinic acid and isopropyl ester of pipecolinic acid. The (R) -pipecolinic acid isopropyl ester was found to be enantiomerically pure and in a yield of 88% based on the (RS) -pipecolinic acid isopropyl ester introduced. Enantiomerically pure (R) -pipecolinic acid was found in low percentages as a by-product.

After separating the biomass by centrifugation (20 min at 8000 * g), the aqueous solution was concentrated in vacuo to a third, the pH was adjusted to 10 with 30% sodium hydroxide and immediately extracted three times with 20 ml each of diethyl ether. The combined organic phases were dried and concentrated with sodium sulfate. By the addition of hydrochloric acid, 0.195 g of (R) -pipecolinic acid isopropyl ester crystallized out (yield 80%, ee> 98% lt. GC).

Example 7

Production of (R) -pipecolinic acid from (RS) -pipecolinic acid isopropyl ester

In a 500 ml conical flask with bullying, add 100 ml of minimal medium 3 and (RS) -pipecolinic acid isopropyl ester

- 14 00 00 ♦ 0 · 0 0 0 0 0 0 · 0 0 0 • 000 0000 00 at a concentration of 4 g / l was added by Pseudomonas putida DSM 9923. The contents were incubated at 30 ° C on a shaker at 140 rpm. After 7.5 hours at OD650 1.4 a sample was taken and incubated without air. After 22 hours, the cell-free supernatant was assayed by GC for content and enantiomeric excess of pipecolinic acid and isopropyl ester of pipecolinic acid. (R) -pipecolinic acid was found to be enantiomerically pure and 80% yield based on the (R) -pipecolinic acid isopropyl ester introduced. An enantiomerically pure (R) -pipecolinic acid isopropyl ester was found as a by-product in a low percentage.

Example 8

Reduction of (R) -pipecolinic acid amide to (R) -2- (aminomethyl) piperidine

11.8 g of lithium aluminum hydride in 500 ml of diethyl ether were suspended in a 2 L flask under nitrogen. 20 g of (R) -pipecolinic acid amide were slowly added at 5 ° C (see Example 3). The mixture was stirred at room temperature for 3 hours and then at reflux for 5 hours. After cooling to room temperature, 12 ml of water, 12 ml of 15% sodium hydroxide, and 36 ml of water were gradually added dropwise. The mixture was stirred at room temperature until it became snow white. 120 g of sodium sulfate was added and stirred at room temperature overnight. The solid was filtered off and washed four times with 250 ml of diethyl ether each time. The combined organic phases were concentrated in vacuo. 12.5 g of (R) -2- (aminomethyl) piperidine were obtained as a yellowish oil (content: 66% by GC). Distillation at 52-53 ° C / 1.8 kPa gave 3.6 g (20%) of (R) -2- (aminomethyl) piperidine pure according to GC as a colorless liquid.

Example 9

Reduction of (R) -pipecolinic acid isopropyl ester to (R) -2- (hydroxymethyl) piperidine

In a 250 ml flask 0.2 g of lithium aluminum hydride in 20 ml of diethyl ether was suspended under a nitrogen atmosphere. At 5 ° C • 9

9 9

9

9 «

9 ««

9 9 ♦ 9

9 9

9999 99 • 99 99 9999 slowly added 90 mg of (R) -pipecolinic acid isopropyl ester hydrochloride. The mixture was stirred at room temperature for 1 hour and then at reflux for 4 hours. After cooling to room temperature, 0.2 ml of water, 0.2 ml of 15% sodium hydroxide and 0.4 ml of water were gradually added dropwise. The mixture was stirred at room temperature until it was snow white. 2 g of sodium sulfate were added and the mixture was stirred at room temperature overnight. The solid was filtered off and washed four times with 20 ml of diethyl ether each time. The combined organic phases were concentrated in vacuo. 47 mg of (R) -2- (hydroxymethyl) piperidine were obtained as a slightly yellow oil (content: 95%, ee> 97% by GC).

Claims (8)

Claims adjusted claims A process for the preparation of (2R) -piperidine derivatives of the general formula I (I) wherein R ¹ is hydroxy, amino or C 1-6 -alkoxy, characterized in that to (RS) -piperidine derivatives of the general formula II: R ² represents hydroxy, amino or C1_g alkoxy and R ¹ and R ² may be the same or different with the proviso that when R ¹ is amino, R ² is not hydroxy or C1_ _6-alkoxy, and when R ¹ is C ₁ _ ₆ - alkoxy, R ² is not hydroxy or amino, treated growing microorganisms which are capable as the sole nitrogen source to utilize α-amides aminokyrboxylových as a racemate or one of its optically active isomers of the formula III (III) wherein a is together with the -NH and -CH optionally substituted 5- or 6-membered saturated heterocyclic ring, in the presence of trace elements and under aerobic conditions, wherein the S-isomer of the (RS) -piperidine II derivative is utilized or degraded as a carbon source and ring cleavage energy. it it - 17 adjusted claims Method according to claim 1, characterized in that (RS) -pipecolinic acid, (RS) -pipecolinic acid ethyl ester, (RS) -pipecolinic acid isopropyl ester or (RS) -pipecolinic acid amide is used as the substrate of formula II. Process according to either of Claims 1 and 2, characterized in that the reaction is carried out with microorganisms of the genus Pseudomonas, in particular of the species Pseudomonas putida. Method according to one of claims 1 to 3, characterized in that the reaction is carried out with microorganisms of the species Pseudo · monas putida (DSM 9923) or functionally equivalent variants or mutants thereof. Method according to one of Claims 1 to 4, characterized in that the addition of the (RS) -pipecolinic acid derivative of the formula II is carried out in such a way that the concentration does not exceed 30% by weight. Process according to one of Claims 1 to 5, characterized in that the reaction is carried out at a pH of from 4 to 9 and at a temperature of from 15 to 80 ° C. A process for preparing (R) -pipecolinic acid of the formula IV OH (IV) characterized in that the (RS) -pipecoline of general formula V is an acid derivative O, wherein R is amino or C ₁ _ ₆ alkoxy-like substrate with growing microorganisms which are capable of utilizing amides and-aminocarboxylic acids in the form of the racemate or one of its optically active isomers of the formula III ·· 41 • 4 »4 4 4 « 4 4 4 4 4 * Wherein A represents together with -NH and -CH an optionally substituted 5- or 6-membered saturated heterocyclyl ring, as the sole nitrogen source, in the presence of trace elements and under aerobic conditions, wherein the reaction is carried out up to to the accumulation of the corresponding (R) -pipecolinic acid derivative of formula VI .0 (VI) R ³ wherein R ^{3 is} as hereinbefore defined, which may optionally be isolated under aerobic conditions, whereby the S-isomer of the (RS) -piperidine V derivative is utilized or degraded as a source of carbon and energy by ring cleavage and then stops supplying oxygen. A process for the preparation of (2R) -piperidone derivatives of the general formula VII (VII), R 'wherein R ⁴ is hydroxy or amino, characterized in that, in the first step, an (RS) -pipecolinic acid derivative of the general formula V • 9 9 9 4 4 • 9 9 9 9 9 9 9 9 9 9 9 9 94 94 9 4 »9 • 9 99 9 4 99 9 wherein R ³ is amino or C ₁ _ _6-alkoxy, as the substrate is converted by growing microorganisms which are capable of utilizing amides, a-aminocarboxylic acids in the form of racemates or one of its optically active isomers of the formula III (III) wherein A represents together -NH and -CH optionally substituted 5- or 6-membered saturated heterocyclyl ring as the sole nitrogen source, in the presence of trace elements and under aerobic conditions under which the S-isomer of the (RS) -piperidine V derivative is utilized or degraded as carbon source and ring cleavage energy to the (R) -pipecolinic acid derivative of formula (VI): R ³ (VI) wherein R ³ is amino or C ₁ _ ₆ -alkoxy, and that in a second step is reduced to the desired compound of formula VII.