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Accueil > Départements > Microbiologie > Marie-Joëlle VIROLLE : Métabolisme Energétique des Streptomyces

Publications de l’équipe

2017



  • C. Esnault, T. Dulermo, A. Smirnov, A. Askora, M. David, A. Deniset-Besseau, I. - B. Holland, et M. - J. Virolle, « Strong antibiotic production is correlated with highly active oxidative metabolism in Streptomyces coelicolor M145 », Scientific Reports, vol. 7, nᵒ 1, 2017.
    Mots-clés : DIR, MESMIC, MICROBIO, SWARM.

  • C. Esnault, D. Leiber, C. Toffano-Nioche, Z. Tanfin, et M. - J. Virolle, « Another example of enzymatic promiscuity: the polyphosphate kinase of Streptomyces lividans is endowed with phospholipase D activity », Applied Microbiology and Biotechnology, vol. 101, nᵒ 1, p. 139-145, 2017.
    Résumé : Polyphosphate kinases (PPK) from different bacteria, including that of Streptomyces lividans, were shown to contain the typical HKD motif present in phospholipase D (PLD) and showed structural similarities to the latter. This observation prompted us to investigate the PLD activity of PPK of S. lividans, in vitro. The ability of PPK to catalyze the hydrolysis of phosphatidylcholine (PC), the PLD substrate, was assessed by the quantification of [(3)H]phosphatidic acid (PA) released from [(3)H]PC-labeled ELT3 cell membranes. Basal cell membrane PLD activity as well as GTPγS-activated PLD activity was higher in the presence than in absence of PPK. After abolition of the basal PLD activity of the membranes by heat or tryptic treatment, the addition of PPK to cell membranes was still accompanied by an increased production of PA demonstrating that PPK also bears a PLD activity. PLD activity of PPK was also assessed by the production of choline from hydrolysis of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) in the presence of the Amplex Red reagent and compared to two commercial PLD enzymes. These data demonstrated that PPK is endowed with a weak but clearly detectable PLD activity. The question of the biological signification, if any, of this enzymatic promiscuity is discussed.
    Mots-clés : Amino Acid Motifs, Cell Membrane, Choline, DBG, eBio, Hydrolysis, Lipid droplets, MESMIC, MICROBIO, PF, Phosphatidic Acids, Phosphatidylcholines, Phospholipase D, Phosphotransferases (Phosphate Group Acceptor), Polyphosphate kinase, Promiscuous enzyme, Protein Conformation, SSFA, Streptomyces lividans.

  • A. Millan-Oropeza, C. Henry, M. Blein-Nicolas, A. Aubert-Frambourg, F. Moussa, J. Bleton, et M. - J. Virolle, « Quantitative Proteomics Analysis Confirmed Oxidative Metabolism Predominates in Streptomyces coelicolor versus Glycolytic Metabolism in Streptomyces lividans », Journal of Proteome Research, 2017.
    Résumé : Recent physiological studies indicated that S. lividans metabolism was mainly glycolytic, whereas S. coelicolor metabolism was mainly oxidative. To determine whether such metabolic characteristics were correlated with consistent proteomics features, a comparative label-free, shotgun proteomics analysis of these strains was carried out. Among 2024 proteins identified, 360 showed significant differences in abundance between the strains. This study revealed that S. coelicolor catabolized glucose less actively than S. lividans, whereas the amino acids present in the medium were catabolized less actively by S. lividans than by S. coelicolor. The abundance of glycolytic proteins in S. lividans was consistent with its high glycolytic activity, whereas the abundance of proteins involved in the catabolism of amino acids in S. coelicolor provided an explanatory basis for its predominantly oxidative metabolism. In this study, conducted under conditions of low O2 availability, proteins involved in resistance to oxidative stress and those belonging to a DosR-like dormancy regulon were abundant in S. coelicolor, whereas tellurium resistance proteins were abundant in S. lividans. This indicated that the strains reacted differently to O2 limitation. Proteins belonging to the CDA, RED, and ACT pathways, usually highly expressed in S. coelicolor, were not detected under these conditions, whereas proteins of siderophores, 5-hydroxyectoine, and terpenoid biosynthetic pathways were present.
    Mots-clés : dormancy regulon, glycolytic metabolism, Mass Spectrometry, MESMIC, MICROBIO, oxidative metabolism, oxidative stress, Secondary metabolites, shotgun label-free quantitative proteomics, Streptomyces, tellurium resistance proteins.

  • A. Millan-Oropeza, R. Rebois, M. David, F. Moussa, A. Dazzi, J. Bleton, M. - J. Virolle, et A. Deniset-Besseau, « Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) for Rapid Determination of Microbial Cell Lipid Content: Correlation with Gas Chromatography-Mass Spectrometry (GC-MS) », Applied Spectroscopy, p. 3702817709459, 2017.
    Résumé : There is a growing interest worldwide for the production of renewable oil without mobilizing agriculture lands; fast and reliable methods are needed to identify highly oleaginous microorganisms of potential industrial interest. The aim of this study was to demonstrate the relevance of attenuated total reflection (ATR) spectroscopy to achieve this goal. To do so, the total lipid content of lyophilized samples of five Streptomyces strains with varying lipid content was assessed with two classical quantitative but time-consuming methods, gas chromatography-mass spectrometry (GC-MS) and ATR Fourier transform infrared (ATR FT-IR) spectroscopy in transmission mode with KBr pellets and the fast ATR method, often questioned for its lack of reliability. A linear correlation between these three methods was demonstrated allowing the establishment of equations to convert ATR values expressed as CO/amide I ratio, into micrograms of lipid per milligram of biomass. The ATR method proved to be as reliable and quantitative as the classical GC-MS and FT-IR in transmission mode methods but faster and more reproducible than the latter since it involves far less manipulation for sample preparation than the two others. Attenuated total reflection could be regarded as an efficient fast screening method to identify natural or genetically modified oleaginous microorganisms by the scientific community working in the field of bio-lipids.
    Mots-clés : ATR FT-IR, attenuated total reflection Fourier transform, Biofuel, gas chromatography–mass spectrometry, GC-MS, infrared spectroscopy, lipid quantification, MESMIC, MICROBIO, Streptomyces, triacylglycerol.


  • L. Yan, Q. Zhang, M. - J. Virolle, et D. Xu, « In conditions of over-expression, WblI, a WhiB-like transcriptional regulator, has a positive impact on the weak antibiotic production of Streptomyces lividans TK24 », PLOS ONE, vol. 12, nᵒ 3, p. e0174781, mars 2017.

2016


  • I. Boukhris, T. Dulermo, H. Chouayekh, et M. - J. Virolle, « Evidence for the negative regulation of phytase gene expression in Streptomyces lividans and Streptomyces coelicolor », Journal of Basic Microbiology, vol. 56, nᵒ 1, p. 59-66, 2016.
    Résumé : Sco7697, a gene encoding a phytase, enzyme able to degrade phytate (myo-inositol 1,2,3,4,5,6-hexakis phosphate), the most abundant phosphorus storing compound in plants is present in the genome of S. coelicolor, a soil born bacteria with a saprophytic lifestyle. The expression of this gene was previously shown to be induced in conditions of Pi limitation by the response regulator PhoP binding to an operator sequence, the PHO box, located upstream of the -35 promoter sequence. A close examination of the promoter region of sco7697 revealed the presence of another putative operator site, a Direct Repeat (DR), located downstream of the -10 promoter sequence. In order to determine whether this DR played a role in regulation of sco7697 expression, different variants of the phytase gene promoter region were transcriptionally fused to the ß-glucuronidase reporter gene (GUS). As expected, deletion of the PHO box led to abolition of sco7697 induction in conditions of Pi limitation. Interestingly, alteration of the DR correlated with a dramatic increase of GUS expression but only when PhoP was present. These results demonstrated that this DR is the site of strong negative regulation by an unknown repressor. The latter would impede the necessary activation of phytase expression by PhoP.
    Mots-clés : 6-Phytase, Bacterial Proteins, Base Sequence, Beta-glucuronidase reporter gene, Gene Expression Regulation, Bacterial, Glucuronidase, MESMIC, MICROBIO, Negative, Operon, PHO box, PhoP, PhoR, Phosphate limitation, Phytic Acid, Positive regulation, Promoter Regions, Genetic, Protein Binding, Repetitive Sequences, Nucleic Acid, Sequence Deletion, Soil Microbiology, Streptomyces coelicolor, Streptomyces lividans, Streptomyces phytase.

  • I. Boukhris, A. Farhat-Khemakhem, K. Bouchaala, M. - J. Virolle, et H. Chouayekh, « Cloning and characterization of the first actinomycete β-propeller phytase from Streptomyces sp. US42 », Journal of Basic Microbiology, vol. 56, nᵒ 10, p. 1080-1089, 2016.
    Résumé : A gene encoding an extracellular phytase was cloned for the first time from an Actinomycete, Streptomyces sp. US42 and sequenced. The sequence of this gene revealed an encoded polypeptide (PHY US42) exhibiting one and six residues difference with the putative phytases of Streptomyces lividans TK24 and Streptomyces coelicolor A3(2), respectively. The molecular modeling of PHY US42 indicated that this phytase belongs to the group of β-propeller phytases that are usually calcium-dependent. PHY US42 was purified and characterized. Its activity was calcium-dependent and maximal at pH 7 and 65 °C. The enzyme was perfectly stable at pH ranging from 5 to 10 and its thermostability was greatly enhanced in the presence of calcium. Indeed, PHY US42 maintained 80% of activity after 10 min of incubation at 75 °C in the presence of 5 mM CaCl2 . PHY US42 was also found to exhibit high stability after incubation at 37 °C for 1 h in the presence of bovine bile and digestive proteases like of pepsin, trypsin, and chymotrypsin. Considering its biochemical properties, PHY US42 could be used as feed additive in combination with an acid phytase for monogastric animals.
    Mots-clés : 6-Phytase, Amino Acid Sequence, Base Sequence, Calcium, Calcium-dependent, Cloning, Molecular, DNA, Fungal, Enzyme Stability, Feed additive, MESMIC, MICROBIO, Sequence Analysis, DNA, Streptomyces, Substrate Specificity, β-propeller phytase.


  • T. Dulermo, F. Coze, M. - J. Virolle, V. Méchin, S. Baumberger, et M. Froissard, « Bioconversion of agricultural lignocellulosic residues into branched-chain fatty acids using <i>Streptomyces lividans</i> », OCL, vol. 23, nᵒ 2, p. A202, 2016.


  • M. - J. Virolle, A. Millan-Oropeza, C. Esnault, A. Smirnov, T. Dulermo, et A. Askora, « In Streptomyces, the switch between primary and secondary metabolism is underpinned by a transition from glycolytic to oxidative metabolism », New Biotechnology, vol. 33, p. S59-S60, 2016.


  • P. Vitry, R. Rebois, E. Bourillot, A. Deniset-Besseau, M. - J. Virolle, E. Lesniewska, et A. Dazzi, « Combining infrared and mode synthesizing atomic force microscopy: Application to the study of lipid vesicles inside Streptomyces bacteria », Nano Research, vol. 9, nᵒ 6, p. 1674-1681, 2016.

2015

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Group MES publications before 2015

Regulation of antibiotics biosynthesis

- Xu D, Waack P, Zhang Q, Werten S, Hinrichs W, Virolle MJ (2014) Structure and regulatory targets of SCO3201, a highly promiscuous TetR-like regulator of Streptomyces coelicolor M145. Biochem Biophys Res Commun. 2014 18 ;450(1):513-8.

- Coze F, Gilard F, Tcherkez G, Virolle MJ, Guyonvarch A (2013) Carbon-Flux distribution within Streptomyces coelicolor metabolism : a comparison between the actinorhodin-producing strain M145 and its non-producing derivative M1146. PlosOne, 23:8 (12) : e84151.

- Seghezzi N, Virolle MJ and Amar P (2013) Novel insights regarding the sigmoidal pattern of resistance to neomycin conferred by the aphII gene, in Streptomyces lividans AMB Express 2013, 3:13

- Le Maréchal P, Decottignies P, Marchand CH, Degrouard J, Jaillard D, Dulermo T, Froissard M, Smirnov A, Chapuis V, Virolle MJ. (2013) Comparative proteomic analysis of the wild-type and the ppk mutant of Streptomyces lividans revealed the importance of storage lipids for antibiotic biosynthesis. Appl Environ Microbiol. vol. 79 no. 19 5907-5917.

- Xu D, Seghezzi N, Esnault C, Virolle MJ. (2010) The Over-expression of a Transcriptional Regulator of the TetR family Represses Antibiotic production and Sporulation in Streptomyces coelicolor.
Appl Environ Microbiol. 76(23):7741-53.

- Ghorbel S, Smirnov A, Chouayekh H, Sperandio B, Esnault C, Kormanec J.and Virolle MJ (2006) Regulation of ppk Expression and In Vivo Function of Ppk in Streptomyces lividans TK24. J Bacteriol. 188(17):6269-76.

- Chouayekh H and Virolle MJ (2002) The polyphosphate kinase is involved in the phosphate control of actinorhodin production in Streptomyces lividans. Mol. Microbiol. 43(4):919-30.)

- Smirnov A, Esnault C, Nowacka-Mazurek A, Martel C, Darbon-Rongère E, Chouayekh H, Ghorbel S, Shikura N, Seghezzi N and Virolle MJ (2008) Toward a better understanding of why and how antibiotic biosynthesis is being triggered in Streptomyces ? Chemical Engineering Transactions. 14 : 47-53. Proceedings of 1rst International Conference on Industrial Biotechnology IBIC 2008. (http://www.aidic.it/IBIC2008/)

- Virolle MJ (2012) Novel antibiotics : some hope ?. Public Service Review : European Union Issue 34 on p 59. Managing Editor Lisa Carnwell.Publishers : Public Service Communications Agency (PSCA) International Ltd. http://www.publicservice.co.uk/.Printer: Pelican Press (Manchester, UK).

- Virolle MJ (2009) The fight against infection. Public Service Review : European Union Issue 17 on p634. ManagingEditor Lisa Carnwell.Publishers : Public Service Communications Agency (PSCA) International Ltd. http://www.publicservice.co.uk/.Printer: Pelican Press (Manchester, UK).

- Virolle MJ (2007) Antibiotic Production in Actinomycetes : Towards a Better Understanding BioforumEurope Oct:37-39. (www.eMagazineBIOforum.com)

Pho regulon

- Lu M, Jiang YL, Wang S, Jin H, Zhang RG, Virolle MJ, Chen Y, Zhou CZ. (2014) Streptomyces coelicolor SCO4226 Is a Nickel Binding Protein. PLoS One. 6 : 9 (10).
Tenconi E, Jourdan S, Motte P, Virolle MJ, Rigali S. (2012) Extracellular sugar phosphates are assimilated by Streptomyces in a PhoP-dependent manner. Antonie

- Van Leeuwenhoek. Antonie Van Leeuwenhoek. 102(3):425-33.
Darbon E, Martel C, Nowacka A, Pegot,S, Moreau PL and Virolle MJ (2012) “Transcriptional and preliminary functional analysis of the six genes cluster located in divergence of phoR/phoP in Streptomyces lividans.” Applied Microbiology and Biotechnology". 95(6):1553-66.

- Ghorbel S, Kormanec J, Artus A and Virolle MJ (2006) Transcriptional studies and regulatory interactions between the phoR-phoP operon and the phoU, mtpA, and ppk genes of Streptomyces lividans TK24. J Bacteriol. 188(2):677-86.

Plant Growth Promoting Rhizobacteria / Streptomyces

- Hamdali H, Virolle MJ, von Jan M, Spröer C, Klenk HP, and Ouhdouch Y (2011) Streptomyces youssoufiensis sp. nov., isolated from a Moroccan phosphate mine. Int. J. Syst. Evol. Microbiol. 61:1104-1108.

- Hamdali H, Smirnov A, Esnault C, Ouhdouch Y and Virolle MJ (2009) Physiological studies and comparative analysis of rock phosphate solubilization abilities of Actinomycetales originating from Moroccan phosphate mines and of Streptomyces lividans Applied Soil Ecology 44(1) : 24-31.

- Hamdali H, Hafidi M, Virolle MJ and Ouhdouch Y. (2008) Growth promotion and protection against damping-off of wheat by two rock phosphate solubilizing Actinomycetes in a P-deficient soil under greenhouse conditions. Applied Soil Ecology, 40 (3) : 510-517. http://top25.sciencedirect.com/subject/agricultural-and-biological-sciences/1/journal/applied-soil-ecology/09291393/archive/20/

- Hamdali H, Hafidi M, Virolle MJ and Ouhdouch Y (2008) Rock phosphate solubilizing Actinomycetes : Screening for their multiple PGPR activities. World Journal of Microbiology and Biotechnology, 24 (11) : 2565-2575.

- Hamdali H, Bouizgarne B, Hafidi M, Lebrihi A, Virolle MJ, Ouhdouch Y (2008) Screening for rock phosphate solubilizing Actinomycetes from Moroccan phosphate Mines. Appl. Soil Ecol., 38(1):12-19.

Others

- Deniset-Besseau A, Prater CB, Virolle MJ, Dazzi A (2014) Monitoring TriAcylGlycerols Accumulation by Atomic Force Microscopy Based Infrared Spectroscopy in Streptomyces Species for Biodiesel Applications. The Journal of Physical Chemistry Letters 654 DOI : 10.1021/jz402393a

- Seghezzi N, Amar P, Koebmann B, Jensen PR, and Virolle MJ (2011) "The construction of a library of synthetic promoters revealed some specific features of strong Streptomyces promoters." Appl Microbiol Biotechnol. 90(2):615-23.

- Chouayekh H and Virolle MJ.(2007) Fate of the sblA transcript in Streptomyces lividans and Escherichia coli. FEMS Microbiol Lett. Nov ; 276(1):42-7.

- Chouayekh H, Nothaft H, Delaunay S, Linder M, Payrastre B, Seghezzi N, Titgemeyer F and Virolle MJ (2007) Phosphoinositide are involved in the control of the growth resumption that follows the transition phase in Streptomyces lividans, in the presence of glucose. J Bacteriol. (189) (3) : 741–49.

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