Cloning and sequence analysis of zooA, a Streptococcus zooepidemicus gene encoding a bacteriocin-like inhibitory substance having a domain structure similar to that of lysostaphin

• Published in: Gene 1997-04, Vol.189 (2), p.255-261
• Main Authors: Simmonds, Robin S, Simpson, Warren J, Tagg, John R
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - isolation & purification; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Bacteriocins - chemistry; Bacteriolysin; Base Sequence; Cloning, Molecular; Endopeptidase; Escherichia coli - genetics; Escherichia coli - metabolism; Genes, Bacterial; Linker; Lysostaphin - chemistry; Mix and match; Modular; Molecular Sequence Data; Peptides; Protein Structure, Tertiary; Recombinant DNA; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Staphylococcus; Streptococcus equi - chemistry; Streptococcus equi - enzymology; Streptococcus equi - genetics
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(96)00859-1

The nucleotide sequence has been determined for zooA, a gene encoding the bacteriocin-like inhibitory substance zoocin A in Streptococcus zooepidemicus strain 4881. The zooA gene product corresponds to the 285-amino acid (aa) zoocin A pre-peptide from which a leader sequence is cleaved to form the 262-aa biologically active molecule of estimated molecular mass 27 877 Da. Expression of zooA in a Gram-negative host was shown by the extracellular release from Escherichia coli, containing cloned zooA, of a biologically active peptide having an identical range of anti-bacterial activity to that of zoocin A, purified from S. zooepidemicus strain 4881. Data base searches revealed sequences having homologies with known muralytic proteins produced by both Gram-positive and Gram-negative bacteria and indicate a `mix and match' blending of domain-type structures, the C-terminal putative receptor-recognition region of the molecule being joined by a threonine-proline-rich linker to an N-terminal putative catalytic region having homology with several known endopeptidases, including lysostaphin.