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Microbiology 150 (2004), 2707-2714; DOI  10.1099/mic.0.27224-0
© 2004 Society for General Microbiology

Expression of bacteriophage {phi}Ea1h lysozyme in Escherichia coli and its activity in growth inhibition of Erwinia amylovora

Won-Sik Kim, Heike Salm and Klaus Geider{dagger}

Max-Planck-Institut für Zellbiologie, Ladenburg, Germany

Correspondence
Klaus Geider
k.geider{at}bba.de

A 3·3 kb fragment from Erwinia amylovora phage {phi}Ea1h in plasmid pJH94 was previously characterized and found to contain an exopolysaccharide depolymerase (dpo) gene and two additional ORFs encoding 178 and 119 amino acids. ORF178 (lyz) and ORF119 (hol) were found to overlap by 19 bp and they resembled genes encoding lysozymes and holins. In nucleotide sequence alignments, lyz had structurally conserved regions with residues important for lysozyme function. The lyz gene was cloned into an expression vector and expressed in Escherichia coli. Active lysozyme was detected only when E. coli cells with the lyz gene and a kanamycin-resistance cassette were grown in the presence of kanamycin. Growth of Erw. amylovora was inhibited after addition of enzyme exceeding a threshold for lysozyme to target cells. When immature pears were soaked in lysates of induced cells, symptoms such as ooze formation and necrosis were retarded or inhibited after inoculation with Erw. amylovora.

{dagger}Present address: Max-Planck-Institut für Zellbiologie, c/o Biologische Bundesanstalt, Schwabenheimer Str. 101, 69221 Dossenheim, Germany.




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[Abstract] [Full Text] [PDF]


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