A5073612200- Bacterial Genetics and Biotechnology
- Enzyme Structure and Function
- Enzyme Production and Characterization
- RNA and protein synthesis mechanisms
- Protein Structure and Dynamics
- Bacteriophages and microbial interactions
- Advanced Fluorescence Microscopy Techniques
- Gene Regulatory Network Analysis
- Bacterial biofilms and quorum sensing
- Protein Hydrolysis and Bioactive Peptides
- Bacillus and Francisella bacterial research
- Phytase and its Applications
- Protein purification and stability
- Toxin Mechanisms and Immunotoxins
- RNA Research and Splicing
- bioluminescence and chemiluminescence research
- CRISPR and Genetic Engineering
- Biochemical and Structural Characterization
- Photosynthetic Processes and Mechanisms
- Fungal and yeast genetics research
- Plant Virus Research Studies
- Plant-Microbe Interactions and Immunity
- Studies on Chitinases and Chitosanases
- Probiotics and Fermented Foods
- Cell Image Analysis Techniques
Inserm
2008-2024
Université de Montpellier
2009-2024
Centre National de la Recherche Scientifique
2009-2024
Centre de Biologie Structurale
2013-2024
Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement
2011-2021
University of Göttingen
2011
Université de Strasbourg
2010
Institut National de Recherche en Santé Publique
2007
Centro de Investigaciones Biológicas Margarita Salas
2007
Laboratoire de Microbiologie et Génétique Moléculaires
1994-2006
Libraries of well-characterised components regulating gene expression levels are essential to many synthetic biology applications. While widely available for the Gram-negative model bacterium Escherichia coli, such libraries lacking Gram-positive Bacillus subtilis, a key organism basic research and biotechnological Here, we engineered genetic toolbox comprising promoters, Ribosome Binding Sites (RBS), protein degradation tags precisely tune in B. subtilis. We first designed modular...
Gram-positive bacteria use a wealth of extracellular signaling peptides, so-called autoinducers, to regulate gene expression according population densities. These “quorum sensing” systems control vital processes such as virulence, sporulation, and transfer. Using x-ray analysis, we determined the structure PlcR, major virulence regulator Bacillus cereus group, obtained mechanistic insights into effects autoinducer binding. Our structural phylogenetic analysis further suggests that all those...
Bacillus licheniformis α-amylase (BLA) is a highly thermostable starch-degrading enzyme that has been extensively studied in both academic and industrial laboratories. For over decade, we have investigated BLA thermal properties identified amino acid substitutions significantly increase or decrease the thermostability. This paper describes cumulative effect of some most beneficial point mutations BLA. Remarkably, Q264S–N265Y double mutation led to rather limited gain stability but improved...
The transcriptional regulator PlcR and its cognate cell-cell signalling peptide PapR form a quorum-sensing system that controls the expression of extra-cellular virulence factors in various species Bacillus cereus group. alleles are clustered into four groups defining pherotypes. However, molecular basis for group specificity remains elusive, largely because biologically relevant is not known. Here, we show vivo active C-terminal heptapeptide precursor, pentapeptide, as previously suggested....
Magnaporthe AVRs and ToxB-like (MAX) effectors constitute a family of secreted virulence proteins in the fungus Pyricularia oryzae (syn . oryzae) , which causes blast disease on numerous cereals grasses. In spite high sequence divergence, MAX share common fold characterized by ß-sandwich core stabilized conserved disulfide bond. this study, we investigated structural landscape diversity within effector repertoire P Combining experimental protein structure determination silico modeling...
Assessing gene expression noise in order to obtain mechanistic insights requires accurate quantification of on many individual cells over a large dynamic range. We used unique method based 2-photon fluorescence fluctuation microscopy measure directly, at the single cell level and with single-molecule sensitivity, absolute concentration fluorescent proteins produced from two Bacillus subtilis promoters that control switch between glycolysis gluconeogenesis. quantified cell-to-cell variations...
This paper provides further understanding of the thermodynamic and structural features determining stability Bacillus licheniformis alpha-amylase (BLA) at two crucial positions, His133 Ala209. Results protein modelling saturated site-directed mutagenesis position 133 209 have been reported in a previous (Declerck et al., 1995, Prot. Engng, 8, 1029-1037). In first part present work, evidence is presented supporting hypothesis that stabilizing mutations reduce rate initial unfolding enzyme...
The transcriptional antiterminator protein LicT regulates the expression of Bacillus subtilis operons involved in beta-glucoside metabolism. It consists an N-terminal RNA-binding domain (co-antiterminator (CAT)) and two phosphorylatable phosphotransferase system regulation domains (PRD1 PRD2). In activated state, each PRD forms a dimeric unit with phosphorylation sites totally buried at dimer interface. Here we present 1.95 A resolution structure inactive PRDs as well molecular solution...
We have identified previously two critical positions for the thermostability of highly thermostable alpha-amylase from Bacillus licheniformis. now introduced all 19 possible amino acid residues to these positions, His133 and Ala209. The most favourable substitutions were Ile Val, respectively, which both increased half-life enzyme at 80 degrees C by a factor approximately 3. At stabilizing effect hydrophobic was observed, although only in case position 133 could clear correlation be drawn...
Mutants affected at the LYS5 locus of Yarrowia lipolytica lack detectable dehydrogenase (SDH) activity. The gene has previously been cloned, and we present here sequence 2.5-kilobase-pair (kb) DNA fragment complementing lys5 mutation. Two large antiparallel open reading frames (ORF1 ORF2) were observed, flanked by potential transcription signals. Both ORFs appear to be transcribed, but several lines evidence suggest that only ORF2 is translated encodes SDH. (i) global amino acid compositions...
The Bacillus subtilis homologous transcriptional antiterminators LicT and SacY control the inducible expression of genes involved in aryl β‐glucoside sucrose utilization respectively. Their RNA‐binding activity is carried by N‐terminal domain (CAT), regulated two similar C‐terminal domains (PRD1 PRD2), which are targets phosphorylation reactions catalysed phosphoenolpyruvate: sugar phosphotransferase system (PTS). In absence corresponding inducer, inactivated BglP, PTS permease (EII)...
A set of 12 Escherichia coli suppressor tRNAs, inserting different amino acids in response to an amber codon, has been used create rapidly numerous protein variants a thermostable amylase; by site-directed mutagenesis, mutations were first introduced into Bacillus licheniformis alpha-amylase gene at position His35, His133, His247, His293, His406, or His450; genes carrying one two then expressed the strains, generating over 100 amylase with predicted acid changes that could be tested for...
The central glycolytic genes repressor (CggR) is a 37 kDa transcriptional protein which plays key role in Bacillus subtilis glycolysis by regulating the transcription of gapA operon. Fructose-1,6-bisphosphate (FBP), identified as effector sugar, has been shown to abolish binding cooperativity CggR its DNA target and modify conformational dynamics CggR/DNA complex. In present study, noncovalent mass spectrometry (MS) was used obtain deeper insights into FBP-dependent interactions. effect FBP...
A sub-lethal hydrostatic pressure (HP) shock of ∼100 MPa elicits a RecA-dependent DNA damage (SOS) response in Escherichia coli K-12, despite the fact that cannot compromise covalent integrity DNA. Prior screens for HP resistance identified Mrr (Methylated adenine Recognition and Restriction), Type IV restriction endonuclease (REase), as instigator this enigmatic HP-induced SOS response. REases tend to target modified sites, E. activity was previously shown be elicited by expression foreign...