期刊
INTERNATIONAL JOURNAL OF MEDICAL MICROBIOLOGY
卷 304, 期 3-4, 页码 269-274出版社
ELSEVIER GMBH
DOI: 10.1016/j.ijmm.2013.10.008
关键词
Escherichia coli 0157:H7; Bacteriophage 933W; Shiga toxin; z1466; 933Wp42; Yfils; Esterase; 5-N-acetyl-9-O-acetyl neuraminic acid; Mucin; Enzyme activity
资金
- program SPP1316 of the Deutsche Forschungsgemeinschaft [Schm1360/4-1]
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 K-m value of about 10 mM and a V-ma(x) 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 (Neu5,9Ac(2)). 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. (C) 2013 Elsevier GmbH. All rights reserved.
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