Location, characterization and expression of lytic enzyme-encoding gene, lytA, of Lactococcus lactis bacteriophage phi US3.
Gene lytA, which encodes lytic enzyme (LytA), of the isometric Lactococcus lactis bacteriophage phi US3, was cloned and expressed in Escherichia coli. The lytA gene was located on the physical map of the phi US3 32-kb DNA that contains cohesive ends. Initial expression of lytA was detected by lysis of an overlay of cells of the phage-sensitive strain, L. lactis SK112. However, LytA appeared to have a broad spectrum and induced lysis in more than 30 different lactococcal strains. The nucleotide sequence of lytA showed a single open reading frame (ORF) of 774 bp encoding a protein of 258 amino acids (aa) with a calculated M(r) of 28,977. This is in agreement with the size of 29 kDa as determined for LytA produced in E. coli using a T7 expression system. The lytA gene is preceded by an ORF that may code for a hydrophobic peptide of 66 aa containing a putative secretion signal, and two putative transmembrane helices. The deduced aa sequence of the phage phi US3 LytA shows similarities to that of the autolysin of Streptococcus pneumoniae which is known to be an amidase.
Department of Biophysical Chemistry, Netherlands Institute for Dairy Research (NIZO), Ede.
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D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004798:Enzymes / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D013294:Lactococcus lactis; D008969:Molecular Sequence Data; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
D004798:Enzymes; D011994:Recombinant Proteins; D009238:N-Acetylmuramoyl-L-alanine Amidase