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The polysaccharide is specifically recognized and degraded by a phage\u2010encoded enzyme, the endo\u2010N\u2010acetylneuraminidase E (Endo NE). Endo NE therefore has become a valuable tool in the study of bacterial pathogenesis and eukaryotic morphogenesis. In this report we describe the molecular cloning of Endo NE and the expression of a functionally active recombinant enzyme. The cloned DNA sequence (2436 bp) encodes a polypeptide of 811 amino acids, which at the 5\u2032 end contains a totally conserved neuraminidase motif. Expressed in <jats:italic>Escherichia coli<\/jats:italic>, the enzyme migrates as a single band of approximately 74 kDa in SDS\u2010PAGE. A central domain of 669 amino acid residues is about 90% homologous to the recently cloned Endo NF. Both phage\u2010induced lysis of bacteria and the catalysis of PSA degradation by the recombinant enzyme are efficiently inhibited by a polyclonal antiserum raised against the intact phage particle. The C\u2010terminal region seems to be essential to enzymatic functions, as truncation of 32 amino acids outside the homology domain completely abolishes Endo NE activity. Our data also indicate that the 38 kDa protein, previously assumed to be a subunit of the Endo NE holoenzyme, is the product of a separate gene locus and is not necessary for <jats:italic>in vitro<\/jats:italic> depolymerase activity.<\/jats:p>","DOI":"10.1111\/j.1365-2958.1995.tb02409.x","type":"journal-article","created":{"date-parts":[[2006,10,28]],"date-time":"2006-10-28T03:17:33Z","timestamp":1162005453000},"page":"441-450","source":"Crossref","is-referenced-by-count":63,"title":["Molecular cloning and functional expression of bacteriophage PK1E\u2010encoded endoneuraminidase Endo NE"],"prefix":"10.1111","volume":"16","author":[{"given":"Rita","family":"Gerardy\u2010Schahn","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Bethe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Brennecke","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martina","family":"M\u00fchlenhoff","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthias","family":"Eckhardt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Ziesing","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Friedrich","family":"Lottspeich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthias","family":"Frosch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2006,10,27]]},"reference":[{"key":"e_1_2_1_2_1","first-page":"4100","article-title":"The 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