Identification and functional characterization of a type I signal peptidase gene of Bacillus megaterium DSM319.
The sipM gene of Bacillus megaterium encoding a type I signal peptidase (SPase) was isolated and structurally characterized. RNA analysis revealed a transcript size in accordance with a bicistronic operon comprising sipM and an adjacent open reading frame. Inactivation of sipM by targeted gene disruption could not be achieved, indicating its essential role for cell viability since there might be no other type I SPase of major importance present in B. megaterium. Plasmid-assisted amplification of the gene resulted in an increase in activity of the heterologous glucanase used as an extracellular reporter, suggesting a potential bottleneck for protein secretion within this species.
Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Corrensstrasse 3, 48149, Münster, Germany.
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Nahrstedt
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Wittchen
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Rachman
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Meinhardt
D000595:Amino Acid Sequence; D001410:Bacillus megaterium / Q000201:enzymology* / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002480:Cellulase / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D017353:Gene Deletion; D018628:Gene Dosage; D015870:Gene Expression; D023061:Gene Order; D005798:Genes, Bacterial; D020043:Genes, Essential; D017930:Genes, Reporter; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009876:Operon; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins; D002480:Cellulase; D012697:Serine Endopeptidases; C026515:type I signal peptidase