Functional analysis of the response regulator DegU in Bacillus megaterium DSM319 and comparative secretome analysis of degSU mutants

Borgmeier, Claudia; Voigt, Birgit; Hecker, Michael; Meinhardt, Friedhelm

doi:10.1007/s00253-011-3302-7

Cited by 8 publications

(1 citation statement)

References 54 publications

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“…In contrast, another gene (vpr) showed discrepancy from the aprE gene in its transcriptional response to salt stress, as transcription was not repressed by salt, at least at some time-points (Figure 4). However, the vpr gene was revealed to be repressed significantly by salt in B. subtilis (Hahne et al, 2010), and also repressed in a DegU-dependent manner in Bacillus megaterium (Borgmeier et al, 2011). In addition, the wprA gene in B. subtilis was expressed during the exponential growth phase (Margot and Karamata, 1996) and was repressed by salt stress (Hahne et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Salt stress represses production of extracellular proteases in Bacillus pumilus

Liu¹,

Huang²,

Hong³

2015

Genet. Mol. Res.

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ABSTRACT.Bacillus pumilus is able to secrete subtilisin-like proteases, one of which has been purified and characterized biochemically, demonstrating great potential for use in industrial applications. In the current study, the biosynthesis and transcription of extracellular proteases in B. pumilus (BA06) under salt stress were investigated using various methods, including a proteolytic assay, zymogram analysis, and real-time PCR. Our results showed that total extracellular proteolytic activity, both in fermentation broth and on milk-containing agar plates, was considerably repressed by salt in a dosage-dependent manner. As Bacillus species usually secret multiple extracellular proteases, a variety of individual extracellular protease encoding genes were selected for real-time PCR analysis. It was shown that proteases encoded by the aprE and aprX genes were the major proteases in the fermentation broth in terms of their transcripts in B. pumilus. Further, transcription of aprE, aprX, and epr genes was indeed repressed by salt stress. In contrast, transcription of other genes (e.g., vpr and wprA) was not repressed or significantly affected by the salt. Conclusively, salt stress represses total extracellular proteolytic activity in B. pumilus, which can largely be ascribed to suppression of the major protease-encoding genes (aprE, R.F. Liu et al. 4940©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (2): 4939-4948 (2015) aprX) at the transcriptional level. In contrast, transcription of other protease-encoding genes (e.g., vpr, wprA) was not repressed by salt stress.

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Section: Discussionmentioning

confidence: 99%

Salt stress represses production of extracellular proteases in Bacillus pumilus

Liu¹,

Huang²,

Hong³

2015

Genet. Mol. Res.

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A Bacillus megaterium System for the Production of Recombinant Proteins and Protein Complexes

Biedendieck

2016

Advanced Technologies for Protein Complex Production and Characterization

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For many years the Gram-positive bacterium Bacillus megaterium has been used for the production and secretion of recombinant proteins. For this purpose it was systematically optimized. Plasmids with different inducible promoter systems, with different compatible origins, with small tags for protein purification and with various specific signals for protein secretion were combined with genetically improved host strains. Finally, the development of appropriate cultivation conditions for the production strains established this organism as a bacterial cell factory even for large proteins. Along with the overproduction of individual proteins the organism is now also used for the simultaneous coproduction of up to 14 recombinant proteins, multiple subsequently interacting or forming protein complexes. Some of these recombinant strains are successfully used for bioconversion or the biosynthesis of valuable components including vitamins. The titers in the g per liter scale for the intra- and extracellular recombinant protein production prove the high potential of B. megaterium for industrial applications. It is currently further enhanced for the production of recombinant proteins and multi-subunit protein complexes using directed genetic engineering approaches based on transcriptome, proteome, metabolome and fluxome data.

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Transcriptome profiling of degU expression reveals unexpected regulatory patterns in Bacillus megaterium and discloses new targets for optimizing expression

Borgmeier

Biedendieck

Hoffmann

et al. 2011

Appl Microbiol Biotechnol

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The first whole transcriptome assessment of a Bacillus megaterium strain provides unanticipated insights into the degSU regulon considered to be of central importance for exo-enzyme production. Regulatory patterns as well as the transcription of degSU itself deviate from the model organism Bacillus subtilis; the number of DegU-regulated secretory enzymes is rather small. Targets for productivity optimization, besides degSU itself, arise from the unexpected DegU-dependent induction of the transition-state regulator AbrB during exponential growth. Induction of secretion-assisting factors, such as the translocase subunit SecY or the signal peptidase SipM, promote hypersecretion. B. megaterium DegSU transcriptional control is advantageous for production purposes, since the degU32 constitutively active mutant conferred hypersecretion of a heterologous Bacillus amyloliquefaciens amylase without the detrimental rise, as for B. subtilis and Bacillus licheniformis, in extracellular proteolytic activities.

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Functional analysis of the response regulator DegU in Bacillus megaterium DSM319 and comparative secretome analysis of degSU mutants

Cited by 8 publications

References 54 publications

Salt stress represses production of extracellular proteases in Bacillus pumilus

Salt stress represses production of extracellular proteases in Bacillus pumilus

A Bacillus megaterium System for the Production of Recombinant Proteins and Protein Complexes

Transcriptome profiling of degU expression reveals unexpected regulatory patterns in Bacillus megaterium and discloses new targets for optimizing expression

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