Collect. Czech. Chem. Commun. 1992, 57, 2187-2191

Purification of Penicillin Amidohydrolase, an Enzyme for Semisynthetic Procedures

Jiří Jiráček, Tomislav Barth, Jiří Velek, Ivo Bláha, Jan Pospíšek and Ivan Svoboda

Institute of Organic Chemistry and Biochemistry, Czechoslovak Academy of Sciences, 166 10 Prague 6


Penicillin amidohydrolase (EC is one of the few enzymes used successfully for deprotection of primary amino groups of semisynthetic peptides. The available material is usually contamined by endo- and exopeptidases. We managed to prepare the enzyme devoid of trypsin- and chymotrypsin-like activities using affinity chromatography with specific ligands: Gly-D-Phe-Phe-Tyr-Thr-Pro-Lys-Thr (the fF peptide) and Leu-Gly-Val-D-Arg-Arg-Gly-Phe (the rR peptide). For further purification of the enzyme affinity chromatography with N-phenylacetyl-D-tert-Leu as a ligand was used.