1887

Abstract

Polyprenol phosphate mannose (PPM) is a lipid-linked sugar donor used by extra-cytoplasmic glycosyl tranferases in bacteria. PPM is synthesiszed by polyprenol phosphate mannose synthase, Ppm1, and in most Actinobacteria is used as the sugar donor for protein O-mannosyl transferase, Pmt, in protein glycosylation. Ppm1 and Pmt have homologues in yeasts and humans, where they are required for protein O-mannosylation. Actinobacteria also use PPM for lipoglycan biosynthesis. Here we show that mutants of have increased susceptibility to a number of antibiotics that target cell wall biosynthesis. The mutants also have mildly increased antibiotic susceptibilities, in particular to β-lactams and vancomycin. Despite normal induction of the vancomycin gene cluster, , the and mutants remained highly vancomycin sensitive indicating that the mechanism of resistance is blocked post-transcriptionally. Differential RNA expression analysis indicated that catabolic pathways were downregulated and anabolic ones upregulated in the mutant compared to the parent or complemented strains. Of note was the increase in expression of fatty acid biosynthetic genes in the mutant. A change in lipid composition was confirmed using Raman spectroscopy, which showed that the mutant had a greater relative proportion of unsaturated fatty acids compared to the parent or the complemented mutant. Taken together, these data suggest that an inability to synthesize PPM () and loss of the glycoproteome ( mutant) can detrimentally affect membrane or cell envelope functions leading to loss of intrinsic and, in the case of vancomycin, acquired antibiotic resistance.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000605
2018-03-01
2020-01-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/164/3/369.html?itemId=/content/journal/micro/10.1099/mic.0.000605&mimeType=html&fmt=ahah

References

  1. Mishra AK, Driessen NN, Appelmelk BJ, Besra GS. Lipoarabinomannan and related glycoconjugates: structure, biogenesis and role in Mycobacterium tuberculosis physiology and host-pathogen interaction. FEMS Microbiol Rev 2011;35:1126–1157 [CrossRef][PubMed]
    [Google Scholar]
  2. Lommel M, Strahl S. Protein O-mannosylation: conserved from bacteria to humans. Glycobiology 2009;19:816–828 [CrossRef][PubMed]
    [Google Scholar]
  3. Fukuda T, Matsumura T, Ato M, Hamasaki M, Nishiuchi Y et al. Critical roles for lipomannan and lipoarabinomannan in cell wall integrity of mycobacteria and pathogenesis of tuberculosis. mBio 2013;4:e00472-12 [CrossRef][PubMed]
    [Google Scholar]
  4. Mishra AK, Alderwick LJ, Rittmann D, Wang C, Bhatt A et al. Identification of a novel α(1—>6) mannopyranosyltransferase MptB from Corynebacterium glutamicum by deletion of a conserved gene, NCgl1505, affords a lipomannan- and lipoarabinomannan-deficient mutant. Mol Microbiol 2008;68:1595–1613 [CrossRef][PubMed]
    [Google Scholar]
  5. Dobos KM, Khoo KH, Swiderek KM, Brennan PJ, Belisle JT. Definition of the full extent of glycosylation of the 45-kilodalton glycoprotein of Mycobacterium tuberculosis. J Bacteriol 1996;178:2498–2506 [CrossRef][PubMed]
    [Google Scholar]
  6. Michell SL, Whelan AO, Wheeler PR, Panico M, Easton RL et al. The MPB83 antigen from Mycobacterium bovis contains O-linked mannose and (1—>3)-mannobiose moieties. J Biol Chem 2003;278:16423–16432 [CrossRef][PubMed]
    [Google Scholar]
  7. Espitia C, Servín-González L, Mancilla R. New insights into protein O-mannosylation in actinomycetes. Mol Biosyst 2010;6:775–781 [CrossRef][PubMed]
    [Google Scholar]
  8. Mahne M, Tauch A, Pühler A, Kalinowski J. The Corynebacterium glutamicum gene pmt encoding a glycosyltransferase related to eukaryotic protein-O-mannosyltransferases is essential for glycosylation of the resuscitation promoting factor (Rpf2) and other secreted proteins. FEMS Microbiol Lett 2006;259:226–233 [CrossRef][PubMed]
    [Google Scholar]
  9. Wehmeier S, Varghese AS, Gurcha SS, Tissot B, Panico M et al. Glycosylation of the phosphate binding protein, PstS, in Streptomyces coelicolor by a pathway that resembles protein O-mannosylation in eukaryotes. Mol Microbiol 2009;71:421–433 [CrossRef][PubMed]
    [Google Scholar]
  10. Gurcha SS, Baulard AR, Kremer L, Locht C, Moody DB et al. Ppm1, a novel polyprenol monophosphomannose synthase from Mycobacterium tuberculosis. Biochem J 2002;365:441–450 [CrossRef][PubMed]
    [Google Scholar]
  11. Rana AK, Singh A, Gurcha SS, Cox LR, Bhatt A et al. Ppm1-encoded polyprenyl monophosphomannose synthase activity is essential for lipoglycan synthesis and survival in mycobacteria. PLoS One 2012;7:e48211 [CrossRef][PubMed]
    [Google Scholar]
  12. Gibson KJ, Eggeling L, Maughan WN, Krumbach K, Gurcha SS et al. Disruption of Cg-Ppm1, a polyprenyl monophosphomannose synthase, and the generation of lipoglycan-less mutants in Corynebacterium glutamicum. J Biol Chem 2003;278:40842–40850 [CrossRef][PubMed]
    [Google Scholar]
  13. Vanderven BC, Harder JD, Crick DC, Belisle JT. Export-mediated assembly of mycobacterial glycoproteins parallels eukaryotic pathways. Science 2005;309:941–943 [CrossRef][PubMed]
    [Google Scholar]
  14. Fernández-Álvarez A, Marín-Menguiano M, Lanver D, Jiménez-Martín A, Elías-Villalobos A et al. Identification of O-mannosylated virulence factors in Ustilago maydis. PLoS Pathog 2012;8:e1002563 [CrossRef][PubMed]
    [Google Scholar]
  15. Gentzsch M, Tanner W. The PMT gene family: protein O-glycosylation in Saccharomyces cerevisiae is vital. EMBO J 1996;15:5752–5759[PubMed]
    [Google Scholar]
  16. Mouyna I, Kniemeyer O, Jank T, Loussert C, Mellado E et al. Members of protein O-mannosyltransferase family in Aspergillus fumigatus differentially affect growth, morphogenesis and viability. Mol Microbiol 2010;76:1205–1221 [CrossRef][PubMed]
    [Google Scholar]
  17. Olson GM, Fox DS, Wang P, Alspaugh JA, Buchanan KL. Role of protein O-mannosyltransferase Pmt4 in the morphogenesis and virulence of Cryptococcus neoformans. Eukaryot Cell 2007;6:222–234 [CrossRef][PubMed]
    [Google Scholar]
  18. Prill SK, Klinkert B, Timpel C, Gale CA, Schröppel K et al. PMT family of Candida albicans: five protein mannosyltransferase isoforms affect growth, morphogenesis and antifungal resistance. Mol Microbiol 2005;55:546–560 [CrossRef][PubMed]
    [Google Scholar]
  19. Willer T, Brandl M, Sipiczki M, Strahl S. Protein O-mannosylation is crucial for cell wall integrity, septation and viability in fission yeast. Mol Microbiol 2005;57:156–170 [CrossRef][PubMed]
    [Google Scholar]
  20. Willer T, Prados B, Falcón-Pérez JM, Renner-Müller I, Przemeck GK et al. Targeted disruption of the Walker-Warburg syndrome gene Pomt1 in mouse results in embryonic lethality. Proc Natl Acad Sci USA 2004;101:14126–14131 [CrossRef][PubMed]
    [Google Scholar]
  21. Cowlishaw DA, Smith MC. Glycosylation of a Streptomyces coelicolor A3(2) cell envelope protein is required for infection by bacteriophage phi C31. Mol Microbiol 2001;41:601–610 [CrossRef][PubMed]
    [Google Scholar]
  22. Cowlishaw DA, Smith MC. A gene encoding a homologue of dolichol phosphate-beta-D-mannose synthase is required for infection of Streptomyces coelicolor A3(2) by phage (phi)C31. J Bacteriol 2002;184:6081–6083 [CrossRef][PubMed]
    [Google Scholar]
  23. MacNeil DJ. Characterization of a unique methyl-specific restriction system in Streptomyces avermitilis. J Bacteriol 1988;170:5607–5612 [CrossRef][PubMed]
    [Google Scholar]
  24. Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA et al. Practical Streptomyces Genetics Norwich: The John Innes Foundation; 2000
    [Google Scholar]
  25. Zhang YM, Rock CO. Membrane lipid homeostasis in bacteria. Nat Rev Microbiol 2008;6:222–233 [CrossRef][PubMed]
    [Google Scholar]
  26. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 2014;15:550 [CrossRef][PubMed]
    [Google Scholar]
  27. Zheng Q, Wang XJ. GOEAST: a web-based software toolkit for Gene Ontology enrichment analysis. Nucleic Acids Res 2008;36:W358–W363 [CrossRef][PubMed]
    [Google Scholar]
  28. Robinson JT, Thorvaldsdóttir H, Winckler W, Guttman M, Lander ES et al. Integrative genomics viewer. Nat Biotechnol 2011;29:24–26 [CrossRef][PubMed]
    [Google Scholar]
  29. Thorvaldsdóttir H, Robinson JT, Mesirov JP. Integrative genomics viewer (IGV): high-performance genomics data visualization and exploration. Brief Bioinform 2013;14:178–192 [CrossRef][PubMed]
    [Google Scholar]
  30. Candeloro P, Grande E, Raimondo R, di Mascolo D, Gentile F et al. Raman database of amino acids solutions: a critical study of extended multiplicative signal correction. Analyst 2013;138:7331–7340 [CrossRef][PubMed]
    [Google Scholar]
  31. Movasaghi Z, Rehman S, Rehman IU. Raman spectroscopy of biological tissues. Applied Spectroscopy Reviews 2007;42:493–541 [CrossRef]
    [Google Scholar]
  32. Walter A, Schumacher W, Bocklitz T, Reinicke M, Rösch P et al. From bulk to single-cell classification of the filamentous growing Streptomyces bacteria by means of Raman spectroscopy. Appl Spectrosc 2011;65:1116–1125 [CrossRef][PubMed]
    [Google Scholar]
  33. Hong HJ, Hutchings MI, Buttner MJ. Biotechnology, Biological Sciences Research Council UK. Vancomycin resistance VanS/VanR two-component systems. Adv Exp Med Biol 2008;631:200–213[Crossref]
    [Google Scholar]
  34. Hong HJ, Hutchings MI, Neu JM, Wright GD, Paget MS et al. Characterization of an inducible vancomycin resistance system in Streptomyces coelicolor reveals a novel gene (vanK) required for drug resistance. Mol Microbiol 2004;52:1107–1121 [CrossRef][PubMed]
    [Google Scholar]
  35. Arthur M, Molinas C, Depardieu F, Courvalin P. Characterization of Tn1546, a Tn3-related transposon conferring glycopeptide resistance by synthesis of depsipeptide peptidoglycan precursors in Enterococcus faecium BM4147. J Bacteriol 1993;175:117–127 [CrossRef][PubMed]
    [Google Scholar]
  36. Bugg TD, Wright GD, Dutka-Malen S, Arthur M, Courvalin P et al. Molecular basis for vancomycin resistance in Enterococcus faecium BM4147: biosynthesis of a depsipeptide peptidoglycan precursor by vancomycin resistance proteins VanH and VanA. Biochemistry 1991;30:10408–10415 [CrossRef][PubMed]
    [Google Scholar]
  37. Healy VL, Lessard IA, Roper DI, Knox JR, Walsh CT. Vancomycin resistance in enterococci: reprogramming of the D-ala-D-Ala ligases in bacterial peptidoglycan biosynthesis. Chem Biol 2000;7:R109–R119 [CrossRef][PubMed]
    [Google Scholar]
  38. Hong HJ, Hutchings MI, Hill LM, Buttner MJ. The role of the novel Fem protein VanK in vancomycin resistance in Streptomyces coelicolor. J Biol Chem 2005;280:13055–13061 [CrossRef][PubMed]
    [Google Scholar]
  39. Bayer AS, Schneider T, Sahl HG. Mechanisms of daptomycin resistance in Staphylococcus aureus: role of the cell membrane and cell wall. Ann N Y Acad Sci 2013;1277:139–158 [CrossRef][PubMed]
    [Google Scholar]
  40. Mascher T, Margulis NG, Wang T, Ye RW, Helmann JD. Cell wall stress responses in Bacillus subtilis: the regulatory network of the bacitracin stimulon. Mol Microbiol 2003;50:1591–1604 [CrossRef][PubMed]
    [Google Scholar]
  41. Boechat AL, Kaihami GH, Politi MJ, Lépine F, Baldini RL. A novel role for an ECF sigma factor in fatty acid biosynthesis and membrane fluidity in Pseudomonas aeruginosa. PLoS One 2013;8:e84775 [CrossRef][PubMed]
    [Google Scholar]
  42. Kingston AW, Subramanian C, Rock CO, Helmann JD. A σW-dependent stress response in Bacillus subtilis that reduces membrane fluidity. Mol Microbiol 2011;81:69–79 [CrossRef][PubMed]
    [Google Scholar]
  43. Hong HJ, Paget MS, Buttner MJ. A signal transduction system in Streptomyces coelicolor that activates the expression of a putative cell wall glycan operon in response to vancomycin and other cell wall-specific antibiotics. Mol Microbiol 2002;44:1199–1211 [CrossRef][PubMed]
    [Google Scholar]
  44. Rida S, Caillet J, Alix JH. Amplification of a novel gene, sanA, abolishes a vancomycin-sensitive defect in Escherichia coli. J Bacteriol 1996;178:94–102 [CrossRef][PubMed]
    [Google Scholar]
  45. Paget MS, Chamberlin L, Atrih A, Foster SJ, Buttner MJ. Evidence that the extracytoplasmic function sigma factor sigmaE is required for normal cell wall structure in Streptomyces coelicolor A3(2). J Bacteriol 1999;181:204–211[PubMed]
    [Google Scholar]
  46. Ashton L, Lau K, Winder CL, Goodacre R. Raman spectroscopy: lighting up the future of microbial identification. Future Microbiol 2011;6:991–997 [CrossRef][PubMed]
    [Google Scholar]
  47. Walter A, Reinicke M, Bocklitz T, Schumacher W, Rösch P et al. Raman spectroscopic detection of physiology changes in plasmid-bearing Escherichia coli with and without antibiotic treatment. Anal Bioanal Chem 2011;400:2763–2773 [CrossRef][PubMed]
    [Google Scholar]
  48. Stöckel S, Kirchhoff J, Neugebauer U, Rösch P, Popp J. The application of Raman spectroscopy for the detection and identification of microorganisms. J Raman Spectrosc 2016;47:89–109 [CrossRef]
    [Google Scholar]
  49. Münchberg U, Rösch P, Bauer M, Popp J. Raman spectroscopic identification of single bacterial cells under antibiotic influence. Anal Bioanal Chem 2014;406:3041–3050 [CrossRef][PubMed]
    [Google Scholar]
  50. Wu H, Volponi JV, Oliver AE, Parikh AN, Simmons BA et al. In vivo lipidomics using single-cell Raman spectroscopy. Proc Natl Acad Sci USA 2011;108:3809–3814 [CrossRef][PubMed]
    [Google Scholar]
  51. Potcoava MC, Futia GL, Aughenbaugh J, Schlaepfer IR, Gibson EA. Raman and coherent anti-Stokes Raman scattering microscopy studies of changes in lipid content and composition in hormone-treated breast and prostate cancer cells. J Biomed Opt 2014;19:111605 [CrossRef][PubMed]
    [Google Scholar]
  52. Nieva C, Marro M, Santana-Codina N, Rao S, Petrov D et al. The lipid phenotype of breast cancer cells characterized by Raman microspectroscopy: towards a stratification of malignancy. PLoS One 2012;7:e46456 [CrossRef][PubMed]
    [Google Scholar]
  53. Hoischen C, Gura K, Luge C, Gumpert J. Lipid and fatty acid composition of cytoplasmic membranes from Streptomyces hygroscopicus and its stable protoplast-type L form. J Bacteriol 1997;179:3430–3436 [CrossRef][PubMed]
    [Google Scholar]
  54. Sandoval-Calderón M, Geiger O, Guan Z, Barona-Gómez F, Sohlenkamp C. A eukaryote-like cardiolipin synthase is present in Streptomyces coelicolor and in most actinobacteria. J Biol Chem 2009;284:17383–17390 [CrossRef][PubMed]
    [Google Scholar]
  55. Guerin ME, Korduláková J, Alzari PM, Brennan PJ, Jackson M. Molecular basis of phosphatidyl-myo-inositol mannoside biosynthesis and regulation in mycobacteria. J Biol Chem 2010;285:33577–33583 [CrossRef][PubMed]
    [Google Scholar]
  56. Gago G, Diacovich L, Arabolaza A, Tsai SC, Gramajo H. Fatty acid biosynthesis in actinomycetes. FEMS Microbiol Rev 2011;35:475–497 [CrossRef][PubMed]
    [Google Scholar]
  57. Singh R, Reynolds KA. Identification and characterization of FabA from the Type II fatty acid synthase of Streptomyces coelicolor. J Nat Prod 2016;79:240–243 [CrossRef][PubMed]
    [Google Scholar]
  58. Feng Y, Cronan JE. Escherichia coli unsaturated fatty acid synthesis: complex transcription of the fabA gene and in vivo identification of the essential reaction catalyzed by FabB. J Biol Chem 2009;284:29526–29535 [CrossRef][PubMed]
    [Google Scholar]
  59. Xiao X, Yu X, Khosla C. Metabolic flux between unsaturated and saturated fatty acids is controlled by the FabA:FabB ratio in the fully reconstituted fatty acid biosynthetic pathway of Escherichia coli. Biochemistry 2013;52:8304–8312 [CrossRef][PubMed]
    [Google Scholar]
  60. Bogdanov M, Dowhan W, Vitrac H. Lipids and topological rules governing membrane protein assembly. Biochim Biophys Acta 2014;1843:1475–1488 [CrossRef][PubMed]
    [Google Scholar]
  61. Mascher T, Hachmann AB, Helmann JD. Regulatory overlap and functional redundancy among Bacillus subtilis extracytoplasmic function sigma factors. J Bacteriol 2007;189:6919–6927 [CrossRef][PubMed]
    [Google Scholar]
  62. Tran TT, Panesso D, Mishra NN, Mileykovskaya E, Guan Z et al. Daptomycin-resistant Enterococcus faecalis diverts the antibiotic molecule from the division septum and remodels cell membrane phospholipids. MBio 2013;4:e00281-13 [CrossRef][PubMed]
    [Google Scholar]
  63. Mishra NN, Bayer AS, Tran TT, Shamoo Y, Mileykovskaya E et al. Daptomycin resistance in enterococci is associated with distinct alterations of cell membrane phospholipid content. PLoS One 2012;7:e43958 [CrossRef][PubMed]
    [Google Scholar]
  64. Kleinschnitz EM, Latus A, Sigle S, Maldener I, Wohlleben W et al. Genetic analysis of SCO2997, encoding a TagF homologue, indicates a role for wall teichoic acids in sporulation of Streptomyces coelicolor A3(2). J Bacteriol 2011;193:6080–6085 [CrossRef][PubMed]
    [Google Scholar]
  65. Helmann JD. Bacillus subtilis extracytoplasmic function (ECF) sigma factors and defense of the cell envelope. Curr Opin Microbiol 2016;30:122–132 [CrossRef]
    [Google Scholar]
  66. Martínez LF, Bishop A, Parkes L, del Sol R, Salerno P et al. Osmoregulation in Streptomyces coelicolor : modulation of SigB activity by OsaC. Mol Microbiol 2009;71:1250–1262 [CrossRef]
    [Google Scholar]
  67. Gordon ND, Ottaviano GL, Connell SE, Tobkin GV, Son CH et al. Secreted-protein response to sigmaU activity in Streptomyces coelicolor. J Bacteriol 2008;190:894–904 [CrossRef][PubMed]
    [Google Scholar]
  68. Hesketh A, Hill C, Mokhtar J, Novotna G, Tran N et al. Genome-wide dynamics of a bacterial response to antibiotics that target the cell envelope. BMC Genomics 2011;12:226 [CrossRef]
    [Google Scholar]
  69. Spirig T, Weiner EM, Clubb RT. Sortase enzymes in Gram-positive bacteria. Mol Microbiol 2011;82:1044–1059 [CrossRef]
    [Google Scholar]
  70. Pallen MJ, Lam AC, Antonio M, Dunbar K. An embarrassment of sortases – a richness of substrates?. Trends Microbiol 2001;9:97–101 [CrossRef]
    [Google Scholar]
  71. Duong A, Capstick DS, di Berardo C, Findlay KC, Hesketh A et al. Aerial development in Streptomyces coelicolor requires sortase activity. Mol Microbiol 2012;83:992–1005 [CrossRef]
    [Google Scholar]
  72. Kim M-S, Dufour YS, Yoo JS, Cho Y-B, Park J-H et al. Conservation of thiol-oxidative stress responses regulated by SigR orthologues in actinomycetes. Mol Microbiol 2012;85:326–344 [CrossRef]
    [Google Scholar]
  73. Strakova E, Zikova A, Vohradsky J. Inference of sigma factor controlled networks by using numerical modeling applied to microarray time series data of the germinating prokaryote. Nucleic Acids Res 2014;42:748–763 [CrossRef]
    [Google Scholar]
  74. Campbell EA, Korzheva N, Mustaev A, Murakami K, Nair S et al. Structural mechanism for rifampicin inhibition of bacterial RNA polymerase. Cell 2001;104:901–912 [CrossRef][PubMed]
    [Google Scholar]
  75. Selinger DW, Saxena RM, Cheung KJ, Church GM, Rosenow C. Global RNA half-life analysis in Escherichia coli reveals positional patterns of transcript degradation. Genome Res 2003;13:216–223 [CrossRef][PubMed]
    [Google Scholar]
  76. Wegrzyn A, Szalewska-Palasz A, Blaszczak A, Liberek K, Wegrzyn G. Differential inhibition of transcription from sigma70- and sigma32-dependent promoters by rifampicin. FEBS Lett 1998;440:172–174[Crossref]
    [Google Scholar]
  77. Bandow JE, Brötz H, Hecker M. Bacillus subtilis tolerance of moderate concentrations of rifampin involves the sigma(B)-dependent general and multiple stress response. J Bacteriol 2002;184:459–467 [CrossRef][PubMed]
    [Google Scholar]
  78. Newell KV, Thomas DP, Brekasis D, Paget MS. The RNA polymerase-binding protein RbpA confers basal levels of rifampicin resistance on Streptomyces coelicolor. Mol Microbiol 2006;60:687–696 [CrossRef][PubMed]
    [Google Scholar]
  79. Lithgow KV, Scott NE, Iwashkiw JA, Thomson EL, Foster LJ et al. A general protein O-glycosylation system within the Burkholderia cepacia complex is involved in motility and virulence. Mol Microbiol 2014;92:116–137 [CrossRef][PubMed]
    [Google Scholar]
  80. Zebian N, Merkx-Jacques A, Pittock PP, Houle S, Dozois CM et al. Comprehensive analysis of flagellin glycosylation in Campylobacter jejuni NCTC 11168 reveals incorporation of legionaminic acid and its importance for host colonization. Glycobiology 2016;26:386–397 [CrossRef]
    [Google Scholar]
  81. Rolain T, Bernard E, Beaussart A, Degand H, Courtin P et al. O-glycosylation as a novel control mechanism of peptidoglycan hydrolase activity. J Biol Chem 2013;288:22233–22247 [CrossRef][PubMed]
    [Google Scholar]
  82. Hugonnet JE, Haddache N, Veckerlé C, Dubost L, Marie A et al. Peptidoglycan cross-linking in glycopeptide-resistant Actinomycetales. Antimicrob Agents Chemother 2014;58:1749–1756 [CrossRef][PubMed]
    [Google Scholar]
  83. Li Y, Florova G, Reynolds KA. Alteration of the fatty acid profile of Streptomyces coelicolor by replacement of the initiation enzyme 3-ketoacyl acyl carrier protein synthase III (FabH). J Bacteriol 2005;187:3795–3799 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000605
Loading
/content/journal/micro/10.1099/mic.0.000605
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error