1887

Abstract

low-molecular-mass (LMM) Penicillin-binding proteins (PBPs) help in hydrolysing the peptidoglycan fragments from their cell wall and recycling them back into the growing peptidoglycan matrix, in addition to their reported involvement in biofilm formation. Biofilms are external slime layers of extra-polymeric substances that sessile bacterial cells secrete to form a habitable niche for themselves. Here, we hypothesize the involvement of LMM PBPs in regulating the nature of exopolysaccharides (EPS) prevailing in its extra-polymeric substances during biofilm formation. Therefore, this study includes the assessment of physiological characteristics of CS109 LMM PBP deletion mutants to address biofilm formation abilities, viability and surface adhesion. Finally, EPS from parent CS109 and its ΔPBP4 and ΔPBP5 mutants were purified and analysed for sugars present. Deletions of LMM PBP reduced biofilm formation, bacterial adhesion and their viability in biofilms. Deletions also diminished EPS production by ΔPBP4 and ΔPBP5 mutants, purification of which suggested an increased overall negative charge compared with their parent. Also, EPS analyses from both mutants revealed the appearance of an unusual sugar, xylose, that was absent in CS109. Accordingly, the reason for reduced biofilm formation in LMM PBP mutants may be speculated as the subsequent production of xylitol and a hindrance in the standard flow of the pentose phosphate pathway.

Funding
This study was supported by the:
  • Science and Engineering Research Board (Award CRG/2019/001558)
    • Principle Award Recipient: AnindyaS Ghosh
  • Department of Biotechnology, Ministry of Science and Technology (Award BT/PR24255/NER/95/716/2017)
    • Principle Award Recipient: S GhoshAnindya
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.001031
2021-02-04
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/167/3/micro001031.html?itemId=/content/journal/micro/10.1099/mic.0.001031&mimeType=html&fmt=ahah

References

  1. Zhang Z, Nadezhina E, Wilkinson KJ. Quantifying diffusion in a biofilm of Streptococcus mutans . Antimicrob Agents Chemother 2011; 55:1075–1081 [View Article][PubMed]
    [Google Scholar]
  2. Beloin C, Roux A, Ghigo JM. Escherichia coli biofilms. Curr Top Microbiol Immunol 2008; 322:249–289 [View Article][PubMed]
    [Google Scholar]
  3. Garrett TR, Bhakoo M, Zhang Z. Bacterial adhesion and biofilms on surfaces. Prog Nat Sci 2008; 18:1049–1056 [View Article]
    [Google Scholar]
  4. Limoli DH, Jones CJ, Wozniak DJ, Cruz S. Bacterial extracellular polysaccharides in biofilm formation and function. Microbiol Spectr 2015; 3:1–30 [View Article][PubMed]
    [Google Scholar]
  5. Donlan RM. Biofilms: microbial life on surfaces. Emerg Infect Dis 2002; 8:881–890 [View Article][PubMed]
    [Google Scholar]
  6. Karatan E, Watnick P. Signals, regulatory networks, and materials that build and break bacterial biofilms. Microbiol Mol Biol Rev 2009; 73:310–347 [View Article][PubMed]
    [Google Scholar]
  7. Vogeleer P, Tremblay YDN, Mafu AA, Jacques M, Harel J. Life on the outside: role of biofilms in environmental persistence of Shiga-toxin producing Escherichia coli . Front Microbiol 2014; 5:317 [View Article][PubMed]
    [Google Scholar]
  8. Van Houdt R, Michiels CW. Role of bacterial cell surface structures in Escherichia coli biofilm formation. Res Microbiol 2005; 156:626–633 [View Article][PubMed]
    [Google Scholar]
  9. Kumar MA, Anandapandian KTK, Parthiban K. Production and characterization of exopolysaccharides (Eps) from biofilm forming marine bacterium. Braz. arch. biol. technol. 2011; 54:259–265 [View Article]
    [Google Scholar]
  10. Rzhepishevska O, Hakobyan S, Ruhal R, Gautrot J, Barbero D et al. The surface charge of anti-bacterial coatings alters motility and biofilm architecture. Biomater Sci 2013; 1:589 [View Article][PubMed]
    [Google Scholar]
  11. Renner LD, Weibel DB. Physicochemical regulation of biofilm formation. MRS Bull 2011; 36:347–355 [View Article][PubMed]
    [Google Scholar]
  12. Teh KH, Flint S, Brooks J, Knight G. (editors) Biofilms in the Dairy Industry, 1st ed. West Sussex, UK: John Wiley & Sons, Ltd; 2015
    [Google Scholar]
  13. Liu Y, Yang CH, Li J. Adhesion and retention of a bacterial phytopathogen Erwinia chrysanthemi in biofilm-coated porous media. Environ Sci Technol 2008; 42:159–165 [View Article][PubMed]
    [Google Scholar]
  14. Irie Y, Preston A, Yuk MH. Expression of the primary carbohydrate component of the Bordetella bronchiseptica biofilm matrix is dependent on growth phase but independent of Bvg regulation. J Bacteriol 2006; 188:6680–6687 [View Article][PubMed]
    [Google Scholar]
  15. Jennings LK, Storek KM, Ledvina HE, Coulon C, Marmont LS et al. Pel is a cationic exopolysaccharide that cross-links extracellular DNA in the Pseudomonas aeruginosa biofilm matrix. Proc Natl Acad Sci U S A 2015; 112:11353–11358 [View Article][PubMed]
    [Google Scholar]
  16. Colvin KM, Irie Y, Tart CS, Urbano R, Whitney JC et al. The Pel and Psl polysaccharides provide Pseudomonas aeruginosa structural redundancy within the biofilm matrix. Environ Microbiol 2012; 14:1913–1928 [View Article][PubMed]
    [Google Scholar]
  17. Gallant CV, Daniels C, Leung JM, Ghosh AS, Young KD et al. Common beta-lactamases inhibit bacterial biofilm formation. Mol Microbiol 2005; 58:1012–1024 [View Article][PubMed]
    [Google Scholar]
  18. Ghosh AS, Chowdhury C, Nelson DE. Physiological functions of D-alanine carboxypeptidases in Escherichia coli . Trends Microbiol 2008; 16:309–317 [View Article][PubMed]
    [Google Scholar]
  19. Kumar A, Sarkar SK, Ghosh D, Ghosh AS. Deletion of penicillin-binding protein 1b impairs biofilm formation and motility in Escherichia coli . Res Microbiol 2012; 163:254–257 [View Article][PubMed]
    [Google Scholar]
  20. Kumar A, Mallik D, Pal S, Mallick S, Sarkar S et al. Escherichia coli O8-antigen enhances biofilm formation under agitated conditions. FEMS Microbiol Lett 2015; 362:fnv112 [View Article][PubMed]
    [Google Scholar]
  21. Sandberg ME, Schellmann D, Brunhofer G, Erker T, Busygin I et al. Pros and cons of using resazurin staining for quantification of viable Staphylococcus aureus biofilms in a screening assay. J Microbiol Methods 2009; 78:104–106 [View Article][PubMed]
    [Google Scholar]
  22. Pawar DM, Rossman ML, Chen J. Role of curli fimbriae in mediating the cells of enterohaemorrhagic Escherichia coli to attach to abiotic surfaces. J Appl Microbiol 2005; 99:418–425 [View Article][PubMed]
    [Google Scholar]
  23. Kaiser TDL, Pereira EM, Dos Santos KRN, Maciel ELN, Schuenck RP et al. Modification of the Congo red agar method to detect biofilm production by Staphylococcus epidermidis . Diagn Microbiol Infect Dis 2013; 75:235–239 [View Article][PubMed]
    [Google Scholar]
  24. Bokranz W, Wang X, Tschäpe H, Römling U. Expression of cellulose and curli fimbriae by Escherichia coli isolated from the gastrointestinal tract. J Med Microbiol 2005; 54:1171–1182 [View Article][PubMed]
    [Google Scholar]
  25. Vesterlund S, Paltta J, Karp M, Ouwehand AC. Measurement of bacterial adhesion-in vitro evaluation of different methods. J Microbiol Methods 2005; 60:225–233 [View Article][PubMed]
    [Google Scholar]
  26. Eboigbodin KE, Biggs CA. Characterization of the extracellular polymeric substances produced by Escherichia coli using infrared spectroscopic, proteomic, and aggregation studies. Biomacromolecules 2008; 9:686–695 [View Article][PubMed]
    [Google Scholar]
  27. Bales PM, Renke EM, May SL, Shen Y, Nelson DC. Purification and characterization of biofilm-associated Eps exopolysaccharides from ESKAPE organisms and other pathogens. PLoS One 2013; 8:e67950 [View Article][PubMed]
    [Google Scholar]
  28. Mandal SK, Singh RP, Patel V. Isolation and characterization of exopolysaccharide secreted by a toxic dinoflagellate, Amphidinium carterae Hulburt 1957 and its probable role in harmful algal blooms (HABs). Microb Ecol 2011; 62:518–527 [View Article][PubMed]
    [Google Scholar]
  29. Singh RP, Shukla MK, Mishra A, Kumari P, Reddy CRK et al. Isolation and characterization of exopolysaccharides from seaweed associated bacteria Bacillus licheniformis . Carbohydr Polym 2011; 84:1019–1026 [View Article]
    [Google Scholar]
  30. Seo E-J, Yoo S-H, Oh K-W, Cha J, Lee HG et al. Isolation of an exopolysaccharide-producing bacterium, Sphingomonas sp. CS101, which forms an unusual type of sphingan. Biosci Biotechnol Biochem 2004; 68:1146–1148 [View Article][PubMed]
    [Google Scholar]
  31. DuBois M, Gilles KA, Hamilton JK, Rebers PA, Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem 1956; 28:350–356 [View Article]
    [Google Scholar]
  32. Christensen BE, Kjosbakken J, Smidsrød O. Partial chemical and physical characterization of two extracellular polysaccharides produced by marine, periphytic Pseudomonas sp. strain NCMB 2021. Appl Environ Microbiol 1985; 50:837–845 [View Article][PubMed]
    [Google Scholar]
  33. Blakeney AB, Harris PJ, Henry RJ, Stone BA. A simple and rapid preparation of alditol acetates for monosaccharide analysis. Carbohydr Res 1983; 113:291–299 [View Article]
    [Google Scholar]
  34. Stahl E. editor Thin-Layer Chromatography, 2nd ed. Springer-Verlag Berlin Heidelberg; 1969
    [Google Scholar]
  35. Brunton NP, Gormley TR, Murray B. Use of the alditol acetate derivatisation for the analysis of reducing sugars in potato tubers. Food Chem 2007; 104:398–402 [View Article]
    [Google Scholar]
  36. Mariscal A, Lopez-Gigosos RM, Carnero-Varo M, Fernandez-Crehuet J. Fluorescent assay based on resazurin for detection of activity of disinfectants against bacterial biofilm. Appl Microbiol Biotechnol 2009; 82:773–783 [View Article][PubMed]
    [Google Scholar]
  37. Cerca N, Pier GB, Vilanova M, Oliveira R, Azeredo J. Quantitative analysis of adhesion and biofilm formation on hydrophilic and hydrophobic surfaces of clinical isolates of Staphylococcus epidermidis . Res Microbiol 2005; 156:506–514 [View Article][PubMed]
    [Google Scholar]
  38. Heilmann C, Schweitzer O, Gerke C, Vanittanakom N, Mack D et al. Molecular basis of intercellular adhesion in the biofilm-forming Staphylococcus epidermidis . Mol Microbiol 1996; 20:1083–1091 [View Article][PubMed]
    [Google Scholar]
  39. Olsén A, Herwald H, Wikström M, Persson K, Mattsson E et al. Identification of two protein-binding and functional regions of curli, a surface organelle and virulence determinant of Escherichia coli . J Biol Chem 2002; 277:34568–34572 [View Article][PubMed]
    [Google Scholar]
  40. Y, Lu H, Wang S, Han J, Xiang H et al. An acidic exopolysaccharide from aloarcula hispanica ATCC33960 and two genes responsible for its synthesis. Archaea 2017; 2017:Article ID 5842958 [View Article][PubMed]
    [Google Scholar]
  41. Urbani R, Sist P, Pletikapic G, Misic T, Svetlicic V. Diatom polysaccharides: Extracellular production, isolation and molecular characterization. In Karunaratne DN. editor In the Complex World of Polysaccharides InTech; 2012 pp 345–370
    [Google Scholar]
  42. Llamas I, Mata JA, Tallon R, Bressollier P, Urdaci MC et al. Characterization of the exopolysaccharide produced by Salipiger mucosus A3, a halophilic species belonging to the Alphaproteobacteria, isolated on the Spanish Mediterranean seaboard. Mar Drugs 2010; 8:2240–2251 [View Article][PubMed]
    [Google Scholar]
  43. Jiao Y, Cody GD, Harding AK, Wilmes P, Schrenk M et al. Characterization of extracellular polymeric substances from acidophilic microbial biofilms. Appl Environ Microbiol 2010; 76:2916–2922 [View Article][PubMed]
    [Google Scholar]
  44. Li O, Qian C-D, Zheng D-Q, Wang P-M, Liu Y et al. Two UDP-glucuronic acid decarboxylases involved in the biosynthesis of a bacterial exopolysaccharide in Paenibacillus elgii . Appl Microbiol Biotechnol 2015; 99:3127–3139 [View Article][PubMed]
    [Google Scholar]
  45. Gu X, Lee SG, Bar-Peled M. Biosynthesis of UDP-xylose and UDP-arabinose in Sinorhizobium meliloti 1021: first characterization of a bacterial UDP-xylose synthase, and UDP-xylose 4-epimerase. Microbiology 2011; 157:260–269 [View Article][PubMed]
    [Google Scholar]
  46. Sprenger GA. Genetics of pentose-phosphate pathway enzymes of Escherichia coli K-12. Arch Microbiol 1995; 164:324–330 [View Article][PubMed]
    [Google Scholar]
  47. Sasaki M, Jojima T, Inui M, Yukawa H. Xylitol production by recombinant Corynebacterium glutamicum under oxygen deprivation. Appl Microbiol Biotechnol 2010; 86:1057–1066 [View Article][PubMed]
    [Google Scholar]
  48. Abdul-Lateef LA, Alturaihy SH A. Effected of xylitol on biofilm formation and growth of Streptococcus pneumoniae isolated from children with acute otitis media in Hilla province/Iraq.. Aust J Basic Appl Sci 2016; 10:23–28
    [Google Scholar]
  49. Badet C, Furiga A, Thébaud N. Effect of xylitol on an in vitro model of oral biofilm. Oral Health Prev Dent 2008; 6:337–341[PubMed]
    [Google Scholar]
  50. da Silva SS, Chandel AK. D-Xylitol: Fermentative Production, Application and Commercialization, 1st ed. Springer-Verlag Berlin Heidelberg; 2012
    [Google Scholar]
  51. Ammons MC, Copié V. Mini-Review: lactoferrin: a bioinspired, anti-biofilm therapeutic. Biofouling 2013; 29:443–455 [View Article][PubMed]
    [Google Scholar]
  52. Ammons MCB, Ward LS, Fisher ST, Wolcott RD, James GA. In vitro susceptibility of established biofilms composed of a clinical wound isolate of Pseudomonas aeruginosa treated with lactoferrin and xylitol. Int J Antimicrob Agents 2009; 33:230–236 [View Article][PubMed]
    [Google Scholar]
  53. Denome SA, Elf PK, Henderson TA, Nelson DE, Young KD. Escherichia coli mutants lacking all possible combinations of eight penicillin binding proteins: viability, characteristics, and implications for peptidoglycan synthesis. J Bacteriol 1999; 181:3981–3993 [View Article][PubMed]
    [Google Scholar]
  54. Nelson DE, Young KD. Contributions of PBP 5 and DD-carboxypeptidase penicillin binding proteins to maintenance of cell shape in Escherichia coli . J Bacteriol 2001; 183:3055–3064 [View Article][PubMed]
    [Google Scholar]
  55. Nelson DE, Young KD. Penicillin binding protein 5 affects cell diameter, contour, and morphology of Escherichia coli . J Bacteriol 2000; 182:1714–1721 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.001031
Loading
/content/journal/micro/10.1099/mic.0.001031
Loading

Data & Media loading...

Supplements

Supplementary material 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