Abstract: The invention consists in a nucleotide sequence having the size of (2646) bp, wherein the order of nucleotides is identical to the order of the nucleotide sequence encoding penicillin acylase from Achromobacter sp. CCM 4824 (formerly Comamonas testosteroni CCM 4824), eventually of the fragments of this sequence having the length of at least 150 nucleotides. The sequence can be used in the formation of a DNA construct, eventually the construct having at least one regulatory sequence regulating the expression of the gene and the production of a polypeptide with the penicillin acylase activity. The sequence can form part of a recombinant expression vector, which consists of the above-mentioned construct, promoter, translational start signal, translational and transcriptional stop signal. Further, the invention concerns a recombinant host cell, containing the nucleic acid construct carried by the vector or integrated into the cell chromosome, and the E.