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

Borgmeier, Claudia; Biedendieck, Rebekka; Hoffmann, Kristina; Jahn, Dieter; Meinhardt, Friedhelm

doi:10.1007/s00253-011-3575-x

Cited by 8 publications

(4 citation statements)

References 75 publications

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“…However, the relationship between DegUϳP and antibiotic production is not clear at present. Recently, DegQ was shown to enhance the production of the peptide antibiotics plipastatin and iturin A (35,36). However, the effects of DegQ are only indirect and are mediated via DegU, since DegQ shares no homology with typical transcriptional regulators, i.e., DNA-binding proteins, and DegQ overexpression cannot complement for the loss of DegU in terms of bacillomycin D synthesis.…”

Section: Discussionmentioning

confidence: 99%

Enhanced Control of Cucumber Wilt Disease by Bacillus amyloliquefaciens SQR9 by Altering the Regulation of Its DegU Phosphorylation

Zhang

Wang

et al. 2014

Appl Environ Microbiol

103

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Bacillus amyloliquefaciens strain SQR9, isolated from the cucumber rhizosphere, suppresses the growth of Fusarium oxysporum in the cucumber rhizosphere and protects the host plant from pathogen invasion through efficient root colonization. In the Gram-positive bacterium Bacillus, the response regulator DegU regulates genetic competence, swarming motility, biofilm formation, complex colony architecture, and protease production. In this study, we report that stepwise phosphorylation of DegU in B. amyloliquefaciens SQR9 can influence biocontrol activity by coordinating multicellular behavior and regulating the synthesis of antibiotics. Results from in vitro and in situ experiments and quantitative PCR (qPCR) studies demonstrate the following: (i) that the lowest level of phosphorylated DegU (DegUϳP) (the degQ mutation) impairs complex colony architecture, biofilm formation, colonization activities, and biocontrol efficiency of Fusarium wilt disease but increases the production of macrolactin and bacillaene, and (ii) that increasing the level of DegUϳP by degQ and degSU overexpression significantly improves complex colony architecture, biofilm formation, colonization activities, production of the antibiotics bacillomycin D and difficidin, and efficiency of biocontrol of Fusarium wilt disease. The results offer a new strategy to enhance the biocontrol efficacy of Bacillus amyloliquefaciens SQR9.

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Section: Discussionmentioning

confidence: 99%

Enhanced Control of Cucumber Wilt Disease by Bacillus amyloliquefaciens SQR9 by Altering the Regulation of Its DegU Phosphorylation

Zhang

Wang

et al. 2014

Appl Environ Microbiol

103

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“…Subsequently, RNA was fragmented and hybridized to a custom-made GE 8×15K microarray for analysis of gene expression of G. sulfurreducens (Agilent Technologies, Santa Clara, California, USA) using the gene expression hybridization kit from Agilent (Agilent Technologies, Santa Clara, California, USA) according to the manufacturer’s instructions. Subsequently, the microarray was analysed using the Microarray C Scanner (Agilent Technologies, Santa Clara, California, USA) as described previously (32). Genes with a |logFC| of >1.0 were considered differentially expressed.…”

Section: Methodsmentioning

confidence: 99%

Quantification of microaerobic growth ofGeobacter sulfurreducens

Engel

VorlaÌˆnder

Biedendieck

et al. 2019

Preprint

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20 Geobacter sulfurreducens was originally considered a strict anaerobe. However, this 21 bacterium was later shown to not only tolerate exposure to oxygen but also to use it as 22 terminal electron acceptor. Research performed has so far only revealed the general ability of 23 G. sulfurreducens to reduce oxygen, but the oxygen uptake rate has not been quantified yet, 24 nor has evidence been provided as to how the bacterium achieves oxygen reduction.25 Therefore, microaerobic growth of G. sulfurreducens was investigated here with better 26 defined operating conditions as previously performed and a transcriptome analysis was 27 performed to elucidate possible metabolic mechanisms important for oxygen reduction in G.28 sulfurreducens. The investigations revealed that cell growth with oxygen is possible to the 29 same extent as with fumarate if the maximum specific oxygen uptake rate (sOUR) of 30 95 mg O2 g CDW -1 h -1 is not surpassed. Hereby, the entire amount of introduced oxygen is 31 reduced. When oxygen concentrations are too high, cell growth is completely inhibited and 32 there is no partial oxygen consumption. Transcriptome analysis suggests a menaquinol 33 oxidase to be the enzyme responsible for oxygen reduction. Transcriptome analysis has 34 further revealed three different survival strategies, depending on the oxygen concentration 35 present. When prompted with small amounts of oxygen, G. sulfurreducens will try to escape 36 the microaerobic area; if oxygen concentrations are higher, cells will focus on rapid and 37 complete oxygen reduction coupled to cell growth; and ultimately cells will form protective 38 layers if a complete reduction becomes impossible. The results presented here have important 39 implications for understanding how G. sulfurreducens survives exposure to oxygen. 40 41 42 43 Introduction 44 Geobacter sulfurreducens, a δ-proteobacterium found in subsurface environments, was 45 originally reported to be a strict anaerobe (1). However, it was later shown that this bacterium 46 can grow with oxygen as terminal electron acceptor when 5 % of oxygen or less are added to 47 the headspace of cultivation flasks (2). But this study did not investigate dissolved oxygen 48 concentrations reached in the liquid phase under these conditions (2). Further, the analysis of 49 the genome of G. sulfurreducens revealed several enzymes that could be involved in the 50 reduction of oxygen (3). The expression of many of the corresponding genes is regulated by 51 the RpoS regulon in G. sulfurreducens, which was shown to be inevitable for cell growth with 52 oxygen (4). Part of the RpoS regulon are genes for the cytochrome c oxidase, which is 53 thought to be most likely responsible for cell growth with oxygen, but genes for a cytochrome 54 d ubiquinol oxidase and a rubredoxin-oxygen oxidoreductase have also been found to be 55 regulated by RpoS and encode for enzymes that may play a role in oxygen reduction (3,5).56 The gene for a cytochrome d ubiquinol oxidase has also been found upregulated in Geobacter 5...

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“…In order to counteract the repression, the regulon DegSU transcriptional control promoted the hyper-secretion of a heterologous B. amyloliquefaciens amylase in B. megaterium. 109 high-level expression of microbial enzymes in B. brevis. B. brevis is also an interesting host for heterologous protein production.…”

Section: Strategies For Achieving High-level Expression Of Microbial mentioning

confidence: 99%

How to achieve high-level expression of microbial enzymes

et al. 2013

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Microbial enzymes have been used in a large number of fields, such as chemical, agricultural and biopharmaceutical industries. The enzyme production rate and yield are the main factors to consider when choosing the appropriate expression system for the production of recombinant proteins. Recombinant enzymes have been expressed in bacteria (e.g., Escherichia coli, Bacillus and lactic acid bacteria), filamentous fungi (e.g., Aspergillus) and yeasts (e.g., Pichia pastoris). The favorable and very advantageous characteristics of these species have resulted in an increasing number of biotechnological applications. Bacterial hosts (e.g., E. coli) can be used to quickly and easily overexpress recombinant enzymes; however, bacterial systems cannot express very large proteins and proteins that require post-translational modifications. The main bacterial expression hosts, with the exception of lactic acid bacteria and filamentous fungi, can produce several toxins which are not compatible with the expression of recombinant enzymes in food and drugs. However, due to the multiplicity of the physiological impacts arising from high-level expression of genes encoding the enzymes and expression hosts, the goal of overproduction can hardly be achieved, and therefore, the yield of recombinant enzymes is limited. In this review, the recent strategies used for the high-level expression of microbial enzymes in the hosts mentioned above are summarized and the prospects are also discussed. We hope this review will contribute to the development of the enzyme-related research field.

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

Cited by 8 publications

References 75 publications

Enhanced Control of Cucumber Wilt Disease by Bacillus amyloliquefaciens SQR9 by Altering the Regulation of Its DegU Phosphorylation

Enhanced Control of Cucumber Wilt Disease by Bacillus amyloliquefaciens SQR9 by Altering the Regulation of Its DegU Phosphorylation

Quantification of microaerobic growth ofGeobacter sulfurreducens

How to achieve high-level expression of microbial enzymes

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