OmpT expression and activity increase in response to recombinant chloramphenicol acetyltransferase overexpression and heat shock in E. coli.
The activity of a 35 kDa protease increased in response to induced expression of chloramphenicol acetyltransferase (CAT) in E. coli. This protease was partially purified, extensively characterized, and identified via the use of zymogram gels as the outer membrane protease, OmpT. In experiments targeting the overlap of well-characterized stress responses, OmpT activity was found to increase in response to heat shock but was only minimally affected by amino acid limitation. The largest increase in activity was found after induction of CAT. OmpT expression levels also increased in response to induction of recombinant CAT overexpression and heat shock. This is the first report of increased activity and expression of an outer membrane protease during cytoplasmic overexpression of a recombinant protein.
Gill
R T
RT
Department of Chemical Engineering, University of Maryland, College Park 20742, USA.
DeLisa
M P
MP
Shiloach
M
M
Holoman
T R
TR
Bentley
W E
WE
D001425:Bacterial Outer Membrane Proteins; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D018869:Heat-Shock Response; D010447:Peptide Hydrolases; D018272:Porins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D018272:Porins; D011994:Recombinant Proteins; C499588:ompT protein, E coli; D015500:Chloramphenicol O-Acetyltransferase; D010447:Peptide Hydrolases