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Johannes Bongaerts
; Simon Esser
; Volker Lorbach
; Lóay Al-Momani
; Michael A. Müller
; Dirk Franke
; Christoph Grondal
; Anja Kurutsch
; Robert Bujnicki
; Ralf Takors
; Leon Raeven
; Marcel Wubbolts
; Roel Bovenberg
; Martin Nieger
; Melanie Schürmann
; Natalie Trachtmann
; Stefan Kozak
; Georg A. Sprenger
; Michael Müller
Veronika Maria Deppe
; Stephanie Klatte
; Johannes Bongaerts
; Karl-Heinz Maurer
; Timothy O'Connell
; Friedhelm Meinhardt
Veronika Maria Deppe
; Johannes Bongaerts
; Timothy O'Connell
; Karl-Heinz Maurer
; Friedhelm Meinhardt
Christian Degering
; Thorsten Eggert
; Michael Puls
; Johannes Bongaerts
; Stefan Evers
; Karl-Heinz Maurer
; Karl-Erich Jaeger
Bacillus subtilis and Bacillus licheniformis are widely used for the large-scale industrial production of proteins. These strains can efficiently secrete proteins into the culture medium using the general secretion (Sec) pathway. A characteristic feature of all secreted proteins is their N-terminal signal peptides, which are recognized by the secretion machinery. Here, we have studied the production of an industrially important secreted protease, namely, subtilisin BPN′ from Bacillus amyloliquefaciens. One hundred seventy-three signal peptides originating from B. subtilis and 220 signal peptides from the B. licheniformis type strain were fused to this secretion target and expressed in B. subtilis, and the resulting library was analyzed by high-throughput screening for extracellular proteolytic activity. We have identified a number of signal peptides originating from both organisms which produced significantly increased yield of the secreted protease. Interestingly, we observed that levels of extracellular protease were improved not only in B. subtilis, which was used as the screening host, but also in two different B. licheniformis strains. To date, it is impossible to predict which signal peptide will result in better secretion and thus an improved yield of a given extracellular target protein. Our data show that screening a library consisting of homologous and heterologous signal peptides fused to a target protein can identify more-effective signal peptides, resulting in improved protein export not only in the original screening host but also in different production strains.
Marco Krämer
; Johannes Bongaerts
; Roel Bovenberg
; Susanne Kremer
; Ulrike Müller
; Sonja Orf
; Marcel Wubbolts
; Leon Raeven