Production of a soluble and active MBP-scFv fusion: favorable effect of the leaky tolR strain.
The 6D6 anti-cortisol scFv was prepared as fusion protein with maltose-binding protein (MBP) to increase the amount of soluble product. This fusion was almost completely insoluble when produced in a wild-type strain of Escherichia coli. However, when MBP-scFv fusion was produced in a tolR leaky strain, it was secreted into the culture medium as an active, soluble protein. Production of recombinant proteins in the tolR strain greatly enhances the recovery of active protein and may be a useful system to produce MBP fusion proteins that would normally aggregate when produced in wild-type bacterial strains.
Chames
P
P
Institut de Biologie Structurale et de Microbiologie, Laboratoire d'Ingénierie et Dynamique des Systèmes Membranaires, CNRS-UPR 9027, Marseille, France.
Fieschi
J
J
Baty
D
D
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins / Q000378:metabolism*; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D006854:Hydrocortisone / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000378:metabolism*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism*; D058619:Maltose-Binding Proteins; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D058619:Maltose-Binding Proteins; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; C461599:maltose transport system, E coli; C060794:tolR protein, E coli; D006854:Hydrocortisone