Preparation and characterization of monomers to tetramers of a Collagen-Like domain from Streptococcus pyogenes

Yong Y. Peng, Violet Stoichevska, Linda Howell, Soren Madsen, Jerome A. Werkmeister, Geoff J. Dumsday, John Alan Maurice Ramshaw

Research output: Contribution to journalArticleResearchpeer-review

8 Citations (Scopus)

Abstract

The collagen like domain Scl2 from Streptococcus pyogenes has been proposed as a potential biomedical material. It is non-cytotoxic and non-immunogenic and can be prepared in good yield in fermentation. The Scl2 collagen domain is about a quarter of the length, 234 residues, of the main collagen type, mammalian type I collagen (1014 residues) that is currently used in biomedical devices. In the present study we have made constructs comprising 1 to 4 copies of the Scl2 collagen domain, plus these same constructs with a CysCys sequence at the C-terminal, analogous to that found in mammalian type III collagens. The yields of these constructs were examined from 2 L fermentation studies. The yields of both series declined with increasing size. Circular dichroism showed that the addition of further collagen domains did not lead to a change in the melting temperature compared to the monomer domain. Addition of the CysCys sequence led to a small additional stabilization of about 2-3_C for the monomer construct when the folding (V) domain was present.

Original languageEnglish
Pages (from-to)378-385
Number of pages8
JournalBioengineered Bugs
Volume5
Issue number6
DOIs
Publication statusPublished - 11 Nov 2014
Externally publishedYes

Keywords

  • Collagen
  • Fermentation
  • Melting temperature
  • Protein polymers
  • Recombinant

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