Analysis of the catalytic domain of the lysin of the lactococcal bacteriophage Tuc2009 by chimeric gene assembling.
An active chimeric cell wall lytic enzyme (Tsl) has been constructed by fusing the region coding for the N-terminal half of the lactococcal phage Tuc2009 lysin and the region coding for the C-terminal domain of the major pneumococcal autolysin. The chimeric enzyme exhibited a glycosidase activity capable of hydrolysing choline-containing pneumococcal cell walls. This experimental approach demonstrated that the Tuc2009 lysin possesses a modular structure and further supports the hypothesis that many cell wall lytic enzymes have evolved by the fusion of preexisting catalytic and peptidoglycan-binding domains.
Sheehan
M M
MM
Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
García
J L
JL
López
R
R
García
P
P
D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D001435:Bacteriophages / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism; D017045:Lactococcus / Q000737:chemistry* / Q000201:enzymology / Q000821:virology; D008969:Molecular Sequence Data; D010957:Plasmids; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D013296:Streptococcus pneumoniae / Q000737:chemistry / Q000201:enzymology; D015678:Viral Structural Proteins / Q000737:chemistry / Q000235:genetics*
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C088289:S protein, bacteriophage Tuc2009; D015678:Viral Structural Proteins; D006026:Glycoside Hydrolases; D000581:Amidohydrolases; C019665:amidase