Engineering Bacillus subtilis for the conversion of the antimetabolite 4-hydroxy-l-threonine to pyridoxine

Commichau, Fabian M.; Alzinger, Ariane; Sande, Rafael; Bretzel, Werner; Reuß, Daniel R.; Dormeyer, Miriam; Chevreux, Bastien; Schuldes, Jörg; Daniel, Rolf; Akeroyd, Michiel; Wyss, Markus; Hohmann, Hans‐Peter; Prágai, Zoltán

doi:10.1016/j.ymben.2015.03.007

Cited by 41 publications

(45 citation statements)

References 56 publications

(54 reference statements)

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“…To dissect the functions of ThrR and CodY in controlling the expression of the thrD and hom‐thrCB genes, we assessed the activities of translational hom‐ and thrD ‐ lacZ fusions in the parent strain 168, in the isogenic ΔthrR and ΔcodY mutants, and in the ΔthrR ΔcodY double mutant. In addition to this, we analysed the activities of the mutated P * hom promoters from the ilvA pseudo‐revertant spr8 and from strain BV708 that was adapted to 4‐hydroxy‐L‐threonine (4‐HT R phenotype), a substance known to interfere with threonine biosynthesis (Westley et al ., ; Commichau et al ., ). BV708 had acquired mutations in the bcaP and ybxG amino acid transporter genes and in the P hom promoter.…”

Section: Resultsmentioning

confidence: 97%

“…BV708 had acquired mutations in the bcaP and ybxG amino acid transporter genes and in the P hom promoter. These mutations enable the bacteria to tolerate high amounts of 4‐hydroxy‐L‐threonine (Commichau et al ., ). In the spr8 strain GP349 and in the 4‐HT R mutant strain the nucleotides C26 and T9 were replaced by T and C, respectively, downstream from the P hom promoter.…”

Section: Resultsmentioning

confidence: 97%

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ThrR, a DNA‐binding transcription factor involved in controlling threonine biosynthesis in Bacillus subtilis

Rosenberg

Müller

Lentes

et al. 2016

Molecular Microbiology

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The threonine dehydratase IlvA is part of the isoleucine biosynthesis pathway in the Gram-positive model bacterium Bacillus subtilis. Consequently, deletion of ilvA causes isoleucine auxotrophy. It has been reported that ilvA pseudo-revertants having a derepressed hom-thrCB operon appear in the presence of threonine. Here we have characterized two classes of ilvA pseudo-revertants. In the first class the hom-thrCB operon was derepressed unmasking the threonine dehydratase activity of the threonine synthase ThrC. In the second class of mutants, threonine biosynthesis was more broadly affected. The first class of ilvA pseudo-revertants had a mutation in the Phom promoter (P*hom ), resulting in constitutive expression of the hom-thrCB operon. In the second class of ilvA pseudo-revertants, the thrR gene encoding a putative DNA-binding protein was inactivated, also resulting in constitutive expression of the hom-thrCB operon. Here we demonstrate that ThrR is indeed a DNA-binding transcription factor that regulates the hom-thrCB operon and the thrD aspartokinase gene. DNA binding assays uncovered the DNA-binding site of ThrR and revealed that the repressor competes with the RNA polymerase for DNA binding. This study also revealed that ThrR orthologs are ubiquitous in genomes from the Gram-positive phylum Firmicutes and in some Gram-negative bacteria.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 97%

ThrR, a DNA‐binding transcription factor involved in controlling threonine biosynthesis in Bacillus subtilis

Rosenberg

Müller

Lentes

et al. 2016

Molecular Microbiology

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“…Nevertheless, genome engineering often leads to undesired effects. In particular, altered gene expression and biosynthesis of enzymes can cause accumulation of toxic intermediates, interference with other pathways and changes of the redox state of the cell (Kokura et al, 1999;Hong and Lee, 2002;Ar evalo-Rodr ıguez et al, 2004;McGary et al, 2013;Commichau et al, 2015). These side-effects are major challenges in genome engineering and have to be addressed individually.…”

Section: Tailor-made Organisms By Genome Streamlining and De Novo Genmentioning

confidence: 99%

“…These side-effects are major challenges in genome engineering and have to be addressed individually. The expression levels of operons can be modified with the right choice of promoters and ribosomal binding sites (Guiziou et al, 2016;Rosenberg et al, 2017), and the interference with other pathways can be circumvented by further genome reduction or evolution experiments (Manabe et al, 2011(Manabe et al, , 2013Commichau et al, 2015). However, we still do not fully understand even intensively studied organisms like B. subtilis and E. coli, which in turn precludes the prediction of possible undesirable phenotypes in many cases.…”

Section: Tailor-made Organisms By Genome Streamlining and De Novo Genmentioning

confidence: 99%

The contribution of bacterial genome engineering to sustainable development

Reuß

Commichau

Stülke

2017

Microbial Biotechnology

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SummaryThe United Nations’ Sustainable Development Goals define important challenges for the prosperous development of mankind. To reach several of these goals, among them the production of value‐added compounds, improved economic and ecologic balance of production processes, prevention of climate change and protection of ecosystems, the use of engineered bacteria can make valuable contributions. We discuss the strategies for genome engineering and how they can be applied to meet the United Nations’ goals for sustainable development.

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“…The bio‐degradation of cellulose has been studied extensively, and a number of cellulolytic enzymes have been characterized . Commercial cellulases are produced mainly by fungi; however, bacteria have also been considered as robust and versatile enzyme producers due to their high growth rate, stability under extreme conditions, and multienzyme complexes . Bacillus strains are often used in modern biological fermentation technology, as they are non‐toxic and biologically safe .…”

Section: Introductionmentioning

confidence: 99%

Solid‐state fermentation of Moringa oleifera leaf meal using Bacillus pumilusCICC 10440

Zhang

Huang

Zhao

et al. 2017

J of Chemical Tech & Biotech

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BACKGROUND Moringa leaf is licensed as a new food ingredient and is rich in nutrients. However, owing to its high cellulose content, the nutrients are difficult to digest and absorb, which limits the product development and application of Moringa leaf. RESULTS Four bacterial strains – Trichoderma reesei CICC 2626, Bacillus pumilus CICC 10440, Bacillus subtilis subsp. CICC 21934, and Bacillus sp. CICC 21942 – were chosen to ferment with Moringa leaf powder. The results of solid fermentation and liquid fermentation show that solid‐state fermentation using Bacillus pumilus CICC 10440 in a 1:60 ratio with Moringa leaf powder produced the greatest amount of soluble protein (285 mg g−1, 2.42 times higher than the control) in 24 h. Using a cellulase activity assay and western blotting, it was found that Bacillus pumilus CICC 10440 uses endoglucanase to dissolve the cellulose to release nutrients from the Moringa oleifera leaf. We expanded the fermentation scale of the Moringa oleifera leaf in a 200 L fermenter and produced a 50 kg batch of Moringa oleifera product. CONCLUSION The results showing improvement in the nutrient release of Moringa leaf via microbial fermentation are a promising start and provide a basis for further product development of Moringa leaf. © 2017 Society of Chemical Industry

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Engineering Bacillus subtilis for the conversion of the antimetabolite 4-hydroxy-l-threonine to pyridoxine

Cited by 41 publications

References 56 publications

ThrR, a DNA‐binding transcription factor involved in controlling threonine biosynthesis in Bacillus subtilis

ThrR, a DNA‐binding transcription factor involved in controlling threonine biosynthesis in Bacillus subtilis

The contribution of bacterial genome engineering to sustainable development

Solid‐state fermentation of Moringa oleifera leaf meal using Bacillus pumilusCICC 10440

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