PROCESS FOR PRODUCING N-PROTECTED D-PROLINE DERIVATIVES

摘要

Microorganism (A) is capable of utilising an N-protected proline derivative of formula (I) (in the form of a racemate or one of its optically active isomers) as its sole nitrogen source, its sole carbon source, or as its sole nitrogen and carbon source. R1 = (CH2)2, COOH, or 1-4C alkoxy, aryl or aryloxy (all optionally substituted); R2 = H or =O. Also claimed are: (1) N-acyl-L-proline-acylase (B), which has the following properties: (a) substrate specificity: hydrolyses N-benzyloxycarbonyl-proline, N-benzoyl-L-proline, N-isobutoxycarbonyl-L-proline, N-benzyloxycarbonyl-L-pyroglutamate, N-benzyloxycarbonyl-DL-pipecolinic acid and N-benzyloxycarbonyl-L-alanine; (b) pH optimum: at pH 6.5 /| 0.2; (c) temperature stability: no loss of activity is observed after 6 hours incubation at 43 deg C and pH 6.5; (d) temperature activity: good activity is detectable at 50 deg C and pH 6.5, and (e) influence of inhibitors: benzyl alcohol and N-benzyloxycarbonyl-D-proline act as inhibitors; (2) preparation of an N-protected cyclic D-amino acid derivative of formula (II) and/or a cyclic L-amino acid derivative of formula (III), comprising conversion of a racemic N-protected cyclic amino acid compound of formula (IV) to (III) using the microorganism described in (A) or the cell-free enzyme described in (B), optionally followed by isolation of (III), where compound (II) is produced as a by-product of this reaction, and may be isolated; A completes a 4-6 membered saturated heterocyclic ring; R3 = (CH2)2COOH, or alkyl, alkoxy, aryl or aryloxy (all optionally substituted); (3) preparation of an N-protected aliphatic D-amino acid derivative of formula (V) and/or an aliphatic L-amino acid derivative of formula (VI), comprising conversion of a racemic aliphatic amino acid derivative of formula (VII) to (VI) using the microorganism described in (A) or the cell-free enzyme described in (B), optionally followed by isolation of (VI). Compound (V) is produced as a by-product of this reaction, and may be isolated. R3 = (CH2)2COOH, or alkyl, alkoxy, aryl or aryloxy (all optionally substituted); R4 = H, straight-chain alkyl (optionally substituted) or an ~w-hydroxyalkyl group; R5 = H or optionally substituted straight chain alkyl.