Bacteriophage 933W encodes a functional esterase downstream of the Shiga toxin 2a operon
- Publication Type
- Journal contribution (peer reviewed)
- Authors
- Nübling, S., Eisele, T., Stöber, H., Funk, J., Polzin, S., Fischer, L., Schmidt, H.
- Year of publication
- 2014
- Published in
- Int. J. Med. Microbiol.
- Band/Volume
- 304/
- DOI
- 10.1016/j.ijmm.2013.10.008
- Page (from - to)
- 269-274
In this study, the 1938 bp open reading frame z1466, which is encoded directly downstream the Shiga toxin 2a (Stx2a) operon in E. coli O157:H7 phage 933W was cloned and expressed recombinantly. Purification with Ni-NTA agarose beads with subsequent SDS-PAGE revealed a 68 kDa protein, designated 933Wp42-His. Analysis of 933Wp42-His demonstrated an esterase activity by activity staining of native gels using triacetin as a substrate. Purified 933Wp42-His demonstrated a Km value of about 10 mM and a Vmax value of 1.667 nkat/ml for 4-methylumbelliferyl-acetate (4-MUF-Ac) as a substrate. The enzyme was most active in the pH-range of 7.0–8.0, and at 50 °C. Furthermore, 933Wp42-His was able to hydrolyze acetic acid from mucin, and 5-N-acetyl-9-O-acetyl neuraminic acid (Neu5,9Ac2). This is the first description of an enzymatic activity of the Stx-phage-encoded protein 933Wp42. Its role in substrate utilization during colonization and human infection is discussed.