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- Fachbereich Chemie und Biotechnologie (847) (remove)
L. Sebastian Meyer-Stork
; Jochen Kalbe
; Rolf Kuropka
; S. L. Sauter
; Hartwig Höcker
; Heinz Berndt
Poly(vinyl acetate), PVAc, with a degree of polymerization Xn = 10 was prepared by chain-transfer radical polymerization using carbon tetrachloride and used as oligomeric plasticizer for commercial PVAc. However, the chlorinated chain ends cause a low thermal stability requiring mild Cl/H substitution. The product exhibits high thermal stability and excellent melt-compounding properties. Blends of oligomeric and commercial PVAc show single glass transition temperatures which decrease with higher oligomer content and exhibit small negative deviations from Fox' linear additivity rule. This indicates plasticization and miscibility being mainly due to entropic effects. Injection-moulded thick specimens show ductile behaviour at oligomer contents >10 wt %, while sheets with a thickness of 0.2–0.5 mm appear flexible already at 7.5 wt %. The oxygen permeability coefficients are an order of magnitude lower than those of low-density polyethylene. Due to the sum of their properties, the plasticized sheets present a promising alternative in the preparation of barrier materials.
Design of enzyme reactors as chromatographic columns for racemic resolution of amino acid esters
(1989)
Sandra Wiegand
; Sascha Dietrich
; Robert Hertel
; Johannes Bongaerts
; Stefan Evers
; Sonja Volland
; Rolf Daniel
; Heiko Liesegang
Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis
(2013)
Michael Rachinger
; Melanie Bauch
; Axel Strittmatter
; Johannes Bongaerts
; Stefan Evers
; Karl-Heinz Maurer
; Rolf Daniel
; Wolfgang Liebl
; Heiko Liesegang
; Armin Ehrenreich
The response of Bacillus licheniformis to heat and ethanol stress and the role of the SigB regulon
(2013)
Birgit Voigt
; Rebecca Schroeter
; Britta Jürgen
; Dirk Albrecht
; Stefan Evers
; Johannes Bongaerts
; Karl-Heinz Maurer
; Thomas Schweder
; Michael Hecker
Anja Wilming
; Jens Begemann
; Stefan Kuhne
; Lars Regestein
; Johannes Bongaerts
; Stefan Evers
; Karl-Heinz Maurer
; Jochen Büchs
Sandra Scheele
; Dan Oertel
; Johannes Bongaerts
; Stefan Evers
; Hendrik Hellmuth
; Karl-Heinz Maurer
; Michael Bott
; Roland Freudl
Disruption experiments targeted at the Bacillus licheniformis degSU operon and GFP-reporter analysis provided evidence for promoter activity immediately upstream of degU. pMutin mediated concomitant introduction of the degU32 allele – known to cause hypersecretion in Bacillus subtilis – resulted in a marked increase in protease activity. Application of 5-fluorouracil based counterselection through establishment of a phosphoribosyltransferase deficient Δupp strain eventually facilitated the marker-free introduction of degU32 leading to further protease enhancement achieving levels as for hypersecreting wild strains in which degU was overexpressed. Surprisingly, deletion of rapG – known to interfere with DegU DNA-binding in B. subtilis – did not enhance protease production neither in the wild type nor in the degU32 strain. The combination of degU32 and Δupp counterselection in the type strain is not only equally effective as in hypersecreting wild strains with respect to protease production but furthermore facilitates genetic strain improvement aiming at biological containment and effectiveness of biotechnological processes.
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.