title: Mutations improving production and secretion of extracellular lipase by Burkholderia glumae PG1.
abstract: Burkholderia glumae is a Gram-negative phytopathogenic bacterium known as the causative agent of rice panicle blight. Strain B. glumae PG1 is used for the production of a biotechnologically relevant lipase, which is secreted into the culture supernatant via a type II secretion pathway. We have comparatively analyzed the genome sequences of B. glumae PG1 wild type and a lipase overproducing strain obtained by classical strain mutagenesis. Among a total number of 72 single nucleotide polymorphisms (SNPs) identified in the genome of the production strain, two were localized in front of the lipAB operon and were analyzed in detail. Both mutations contribute to a 100-fold overproduction of extracellular lipase in B. glumae PG1 by affecting transcription of the lipAB operon and efficiency of lipase secretion. We analyzed each of the two SNPs separately and observed a stronger influence of the promoter mutation than of the signal peptide modification but also a cumulative effect of both mutations. Furthermore, fusion of the mutated LipA signal peptide resulted in a 2-fold increase in secretion of the heterologous reporter alkaline phosphatase from Escherichia coli.
authors.affiliation: Institute of Molecular Enzyme Technology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Knapp
authors.affiliation: Institute of Microbiology and Genetics, Department of Genomic and Applied Microbiology and Göttingen Genomics Laboratory, Georg-August University Göttingen, Göttingen, Germany.
authors.forename: Sonja
authors.initials: S
authors.lastname: Voget
authors.affiliation: Biocenter Klein Flottbek, Department of Microbiology and Biotechnology, University of Hamburg, Hamburg, Germany.
authors.forename: Rong
authors.initials: R
authors.lastname: Gao
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland, Helmholtz Centre for Infection Research and Pharmaceutical Biotechnology, Saarland University, Saarbrücken, Germany.
authors.forename: Nestor
authors.initials: N
authors.lastname: Zaburannyi
authors.affiliation: Biocenter Klein Flottbek, Department of Microbiology and Biotechnology, University of Hamburg, Hamburg, Germany.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Krysciak
authors.affiliation: BASF SE, Biocatalysis and Fine Chemicals Research, Ludwigshafen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Breuer
authors.affiliation: BASF SE, Biocatalysis and Fine Chemicals Research, Ludwigshafen, Germany.|Institute of Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Hauer
authors.affiliation: Biocenter Klein Flottbek, Department of Microbiology and Biotechnology, University of Hamburg, Hamburg, Germany.
authors.forename: Wolfgang R
authors.initials: WR
authors.lastname: Streit
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland, Helmholtz Centre for Infection Research and Pharmaceutical Biotechnology, Saarland University, Saarbrücken, Germany.
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
authors.affiliation: Institute of Microbiology and Genetics, Department of Genomic and Applied Microbiology and Göttingen Genomics Laboratory, Georg-August University Göttingen, Göttingen, Germany.
authors.forename: Rolf
authors.initials: R
authors.lastname: Daniel
authors.affiliation: Institute of Molecular Enzyme Technology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany. karl-erich.jaeger@fz-juelich.de.|Forschungszentrum Jülich GmbH, Institute of Bio- and Geosciences IBG-1: Biotechnology, Jülich, Germany. karl-erich.jaeger@fz-juelich.de.
authors.forename: Karl-Erich
authors.initials: KE
authors.lastname: Jaeger
keywords: Burkholderia glumae,; Enzyme production; Lipase,; Secretion,