Surface modification using interfacial assembly of the Streptomyces chaplin proteins


The chaplin proteins are instrumental in the formation of reproductive aerial structures by the filamentous bacterium Streptomyces coelicolor. They lower the water surface tension thereby enabling aerial growth. In addition, chaplins provide surface hydrophobicity to the aerial hyphae by assembling on the cell surface into an amphipathic layer of amyloid fibrils. We here show that mixtures of cell wall-extracted chaplins can be used to modify a variety of hydrophilic and hydrophobic surfaces in vitro thereby changing their nature. Assembly on glass leads to a protein coating that makes the surface hydrophobic. Conversely, the assembly of chaplins on hydrophobic surfaces renders them hydrophilic. Furthermore, we show that chaplins can stabilize emulsions of oil into water and have an unprecedented surface activity at high pH. Interestingly, this high surface activity coincides with the interfacial assembly of chaplins into a semi-liquid membrane, as opposed to the rigid membrane formed at neutral pH. This semi-liquid membrane possibly represents a trapped intermediate in the assembly process towards the more rigid amyloidal conformation. Taken together, our data shows that chaplins are suitable candidate proteins for a wide range of biotechnological applications.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 199

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7


  1. Bilewicz R, Witomski J, Van der Heyden A, Tagu D, Palin B, Rogalska E (2004) Modification of electrodes with self-assembled hydrophobin layers. J Phys Chem B 105(40):9772–9777

  2. Blanco LP, Evans ML, Smith DR, Badtke MP, Chapman MR (2012) Diversity, biogenesis, and function of microbial amyloids. Trends Microbiol 20(2):66–73

  3. Bokhove M, Claessen D, de Jong W, Dijkhuizen L, Boekema EJ, Oostergetel GT (2013) Chaplins of Streptomyces coelicolor self-assemble into two distinct functional amyloids. J Struct Biol. doi:10.1016/j.jsb.2013.08.013, in press

  4. Capstick DS, Jomaa A, Hanke C, Ortega J, Elliot MA (2011) Dual amyloid domains promote differential functioning of the chaplin proteins during Streptomyces aerial morphogenesis. Proc Natl Acad Sci U S A 108(24):9821–9826

  5. Chater KF, Horinouchi S (2003) Signaling early developmental events in two highly diverged Streptomyces species. Mol Microbiol 48(1):9–15

  6. Claessen D, Wösten HAB, van Keulen G, Faber OG, Alves AM, Meijer WG, Dijkhuizen L (2002) Two novel homologous proteins of Streptomyces coelicolor and Streptomyces lividans are involved in the formation of the rodlet layer and mediate attachment to a hydrophobic surface. Mol Microbiol 44(6):1483–1492

  7. Claessen D, Rink R, de Jong W, Siebring J, de Vreugd P, Boersma FG, Dijkhuizen L, Wösten HAB (2003) A novel class of secreted hydrophobic proteins in involved in aerial hyphae formation in Streptomyces coelicolor by forming amyloid-like fibrils. Genes Dev 17(14):1714–1726

  8. Claessen D, Stokroos I, Deelstra HJ, Penninga NA, Bormann C, Salas JA, Dijkhuizen L, Wösten HAB (2004) The formation of the rodlet layer of Streptomycetes is the result of the interplay between rodlins and chaplins. Mol Microbiol 53(2):433–443

  9. Claessen D, de Jong W, Dijkhuizen L, Wösten HAB (2006) Regulation of Streptomyces development: reach for the sky! Trends Microbiol 14(7):313–319

  10. de Jong W, Wösten HAB, Dijkhuizen L, Claessen D (2009) Attachment of Streptomyces coelicolor is mediated by amyloidal fimbriae that are anchored to the cell surface via cellulose. Mol Microbiol 73(6):1128–1140

  11. de Jong W, Vijgenboom E, Dijkhuizen L, Wösten HAB, Claessen D (2012) SapB and the rodlins are required for development of Streptomyces coelicolor in high osmolarity media. FEMS Microbiol Lett 329(2):154–159

  12. Duong A, Capstick DS, Di Berardo C, Findlay KC, Hesketh A, Hong HJ, Elliot MA (2012) Aerial development in Streptomyces coelicolor requires sortase activity. Mol Microbiol 83(5):992–1005

  13. Eisenberg D, Jucker M (2012) The amyloid state of proteins in human diseases. Cell 148(6):1188–1203

  14. Elliot MA, Talbot NJ (2004) Building filaments in the air: aerial morphogenesis in bacteria and fungi. Curr Opin Microbiol 7(6):594–601

  15. Elliot MA, Karoonuthaisiri N, Huang J, Bibb MJ, Cohen SN, Kao CM, Buttner MJ (2003) The chaplins: a family of hydrophobic cell-surface proteins involved in aerial mycelium formation in Streptomyces coelicolor. Genes Dev 17(14):1727–1740

  16. Falabella P, Riviello L, Pascale M, Lelio ID, Tettamanti G, Grimaldi A, Iannone C, Monti M, Pucci P, Tamburro AM, Deeguileor M, Gigliotti S, Pennacchio F (2012) Functional amyloids in insect immune response. Insect Biochem Mol Biol 42(3):203–211

  17. Flärdh K, Buttner MJ (2009) Streptomyces morphogenetics: dissecting differentiation in a filamentous bacterium. Nat Rev Microbiol 7(1):36–49

  18. Fodera V, Groenning M, Vetri V, Librizzi F, Spagnolo S, Cornett C, Olsen L, van der Weert M, Leone M (2008) Thioflavin T hydroxylation at basic pH and its effect on amyloid fibril detection. J Phys Chem 112:15174–15181

  19. Fowler DM, Koulov AV, Alory-Jost C, Marks MS, Balch WE, Kelly JW (2006) Functional amyloid formation within mammalian tissue. PLOS Biol 4(1):e6

  20. Gras SL, Claessen D (2013) Functional amyloid fibrils: lessons from microbes. In: Havlicek V, Spizek J (eds) Natural Products Analysis: instrumentation, Methods, and Applications. Wiley (in press)

  21. Gebbink MFBH, Claessen D, Bouma B, Dijkhuizen L, Wösten HAB (2005) Amyloids—a functional coat for microorganisms. Nat Rev Microbiol 3(4):333–341

  22. Hozzein WN, Ali MIA, Hammouda O, Mousa AS, Goodfellow M (2011) Streptomyces sannurensis sp. nov., a new alkaliphilic member of the genus Streptomyces isolated from Wadi Sannur in Egypt. Afr J Microbiol Res 5(11):1329–1334

  23. Janssen MI, van Leeuwen MB, van Kooten TG, de Vries J, Dijkhuizen L, Wösten HAB (2004) Promotion of fibroblast activity by coating with hydrophobins in the β-sheet end state. Biomaterials 25(14):2731–2739

  24. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685

  25. Li WJ, Zhang YG, Zhang YQ, Tang SK, Xu P, Xu LH, Jiang CL (2005) Streptomyces sodiiphilus sp. Nov., a novel alkaliphilic actinomycete. Int J Syst Evol Microbiol 55(3):1329–1333

  26. Linder M, Szilvay GR, Nakari-Setala T, Soderlund H, Penttila M (2002) Surface adhesion of fusion proteins containing the hydrophobins HFBI and HFBII from Trichoderma reesei. Protein Sci 11(9):2257–2266

  27. Linder MB, Szilvay GR, Nakari-Setala T, Penttila ME (2005) Hydrophobins: the protein-amphiphiles of filamentous fungi. FEMS Microbiol Rev 29(5):877–896

  28. Sawyer EB, Claessen D, Haas M, Hurgobin B, Gras SL (2011) The assembly of individual chaplin peptides from Streptomyces coelicolor into functional amyloid fibrils. PLoS One 6(4):18839–18839

  29. Scholtemeijer K, Janssen MI, van Leeuwen MB, van Kooten TG, Hektor H, Wösten HAB (2004) The use of hydrophobins to functionalize surfaces. Bio-Med Mater Eng 14(4):447–454

  30. Scholtmeijer K, de Vocht ML, Rink R, Robillard GT, Wösten HAB (2009) Assembly of the fungal SC3 hydrophobin into functional amyloid fibrils depends on its concentration and is promoted by cell wall polysaccharides. J Biol Chem 284(39):26309–26314

  31. Srinivasan R, Jones EM, Liu K, Ghiso J, Marchant RE, Zagorski MG (2003) pH-dependent amyloid and protofibril formation by the ABri peptide of familial British dementia. J Mol Biol 333(5):1003–1023

  32. Stauffer CE (1965) The measurement of surface tension by the pendant drop technique. J Phys Chem 69(6):1933–1938

  33. Sunde M, Kwan AHY, Templeton MD, Beever RE, Mackay JP (2008) Structural analysis of hydrophobins. Micron 39(7):773–784

  34. Wösten HAB (2001) Hydrophobins: multipurpose proteins. Annu Rev Microbiol 55:625–646

  35. Xu W, Zhang C, Derreumaux P, Graslund A, Morozova-Roche L, Mu Y (2011) Intrinsic determinants of Abeta12-24 pH-dependent self-assembly revealed by combined computational and experimental studies. PLoS One 6(9):1–12

Download references


We would like to thank Karin Scholtmeijer and Han Wösten for stimulating discussions and their help with the water contact angle and surface tension measurements.

Author information

Correspondence to Dennis Claessen or Eize Stamhuis.

Additional information

David Matthias Ekkers and Dennis Claessen contributed equally to this manuscript.

Electronic supplementary material

Below is the link to the electronic supplementary material.

(WMV 1646 kb)


(PDF 4147 kb)


(WMV 1646 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ekkers, D.M., Claessen, D., Galli, F. et al. Surface modification using interfacial assembly of the Streptomyces chaplin proteins. Appl Microbiol Biotechnol 98, 4491–4501 (2014).

Download citation


  • Chaplins
  • Surface modification
  • Hydrophobins
  • Surfactant
  • Functional amyloid
  • Application