High-level transcription of large gene regions: a novel T(7) RNA-polymerase-based system for expression of functional hydrogenases in the phototrophic bacterium Rhodobacter capsulatus.
High-level synthesis of complex enzymes like bacterial [NiFe] hydrogenases, in general, requires an expression system that allows concerted expression of a large number of genes. So far, it has not been possible to overproduce a hydrogenase in a stable and active form by using a customary expression system. Therefore we started to establish a new, T(7)-based expression system in the phototrophic bacterium Rhodobacter capsulatus. The beneficial properties of this bacterial host in combination with the unique capacity of T(7) RNA polymerase to synthesize long transcripts will allow the high-level synthesis and assembly of active hydrogenase as well as other complex enzymes in the near future.
Drepper
T
T
Institut für Molekulare Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf, Forschungszentrum Jülich, D-52426 Jülich, Germany. t.drepper@tz-juelich.de
Arvani
S
S
Rosenau
F
F
Wilhelm
S
S
Jaeger
K-E
KE
D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D006864:Hydrogenase / Q000235:genetics* / Q000378:metabolism; D012242:Rhodobacter sphaeroides / Q000201:enzymology*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics*
D014764:Viral Proteins; D006864:Hydrogenase; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases