1. Overproducing the BAM complex improves secretion of difficult-to-secrete recombinant autotransporter chimeras
- Author
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Coen Kuijl, Peter van Ulsen, Trang H Phan, Joen Luirink, Dung T Huynh, Wouter S. P. Jong, Molecular Microbiology, AIMMS, LaserLaB - Molecular Biophysics, LaserLaB - Analytical Chemistry and Spectroscopy, Medical Microbiology and Infection Prevention, and AII - Infectious diseases
- Subjects
Type V secretion systems ,Protein domain ,Bioengineering ,Hbp ,Outer membrane proteins ,Microbiology ,Applied Microbiology and Biotechnology ,law.invention ,03 medical and health sciences ,law ,Escherichia coli ,Secretion ,Overproduction ,030304 developmental biology ,0303 health sciences ,BAM complex ,030306 microbiology ,Chemistry ,Research ,Cell Membrane ,Periplasmic space ,Surface display ,QR1-502 ,Recombinant Proteins ,Cell biology ,Protein Transport ,Recombinant DNA ,Protein Translocation Systems ,Cell envelope ,Bacterial outer membrane ,Biotechnology ,Autotransporters - Abstract
Monomeric autotransporters have been used extensively to transport recombinant proteins or protein domains to the cell surface of Gram-negative bacteria amongst others for antigen display. Genetic fusion of such antigens into autotransporters has yielded chimeras that can be used for vaccination purposes. However, not every fusion construct is transported efficiently across the cell envelope. Problems occur in particular when the fused antigen attains a relatively complex structure in the periplasm, prior to its translocation across the outer membrane. The latter step requires the interaction with periplasmic chaperones and the BAM (β-barrel assembly machinery) complex in the outer membrane. This complex catalyzes insertion and folding of β-barrel outer membrane proteins, including the β-barrel domain of autotransporters. Here, we investigated whether the availability of periplasmic chaperones or the BAM complex is a limiting factor for the surface localization of difficult-to-secrete chimeric autotransporter constructs. Indeed, we found that overproduction of in particular the BAM complex, increases surface display of difficult-to-secrete chimeras. Importantly, this beneficial effect appeared to be generic not only for a number of monomeric autotransporter fusions but also for fusions to trimeric autotransporters. Therefore, overproduction of BAM might be an attractive strategy to improve the production of recombinant autotransporter constructs. Supplementary Information The online version contains supplementary material available at 10.1186/s12934-021-01668-2.
- Published
- 2021