Creating a bifunctional protein by insertion of beta-lactamase into the maltodextrin-binding protein.
Hybrid proteins were generated by inserting the penicillin-hydrolyzing enzyme, TEM beta-lactamase (Bla), into the maltodextrin-binding protein (MalE). The inserted Bla was functionally accommodated by MalE when it was placed within permissive sites. The maltose binding and penicillinase activities of purified hybrids were indistinguishable from those of the wild-type MalE and Bla proteins. Moreover, these hybrids displayed an additional unexpected property: maltose stabilized the active site of inserted Bla.
Betton
J M
JM
Département des Biotechnologies, Institut Pasteur-CNRS URA1444, Paris, France. jmbetton@pasteur.fr;mhofnung@pasteur.fr
Jacob
J P
JP
Hofnung
M
M
Broome-Smith
J K
JK
D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D029968:Escherichia coli Proteins; D033902:Periplasmic Binding Proteins; D010957:Plasmids; D017510:Protein Folding; D001618:beta-Lactamases / Q000737:chemistry* / Q000378:metabolism
D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D033902:Periplasmic Binding Proteins; D001618:beta-Lactamases