Generation of metal-binding staphylococci through surface display of combinatorially engineered cellulose-binding domains

H Wernerus, J Lehtio, T Teeri, PA Nygren, S Stahl*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Ni2+-binding staphylococci were generated through surface display of combinatorially engineered variants of a fungal cellulose-binding domain (CBD) from Trichoderma reesei cellulase CeI7A. Novel CBD variants were generated by combinatorial protein engineering through the randomization of 11 amino acid positions, and eight potentially Ni2+-binding CBDs were selected by phage display technology. These new variants were subsequently genetically introduced into chimeric surface proteins for surface display on Staphylococcus carnosus cells. The expressed chimeric proteins were shown to be properly targeted to the cell wall of S. carnosus cells, since full-length proteins could be extracted and affinity purified. Surface accessibility for the chimeric proteins was demonstrated, and furthermore, the engineered CBDs, now devoid of cellulose-binding capacity, were shown to be functional with regard to metal binding, since the recombinant staphylococci had gained Ni2+-binding capacity. Potential environmental applications for such tailor-made metal-binding bacteria as bioadsorbents in biofilters or biosensors are discussed.

Original languageEnglish
Pages (from-to)4678-4684
Number of pages7
JournalApplied and Environmental Microbiology
Volume67
Issue number10
Publication statusPublished - Oct 2001
MoE publication typeA1 Journal article-refereed

Keywords

  • NEUROSPORA-CRASSA METALLOTHIONEIN
  • BACTERIAL RECEPTOR DOMAIN
  • ESCHERICHIA-COLI-CELLS
  • HYICUS SUBSP HYICUS
  • HEAVY-METALS
  • BIOCHEMICAL-PROPERTIES
  • PROTEIN-A
  • PEPTIDES
  • CARNOSUS
  • EXPRESSION

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