Generation of metal-binding staphylococci through surface display of combinatorially engineered cellulose-binding domains
2001 (English)In: Applied and Environmental Microbiology, ISSN 0099-2240, E-ISSN 1098-5336, Vol. 67, no 10, 4678-4684 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
2001. Vol. 67, no 10, 4678-4684 p.
neurospora-crassa metallothionein, bacterial receptor domain, escherichia-coli-cells, hyicus subsp hyicus, heavy-metals, biochemical-properties, protein-a, peptides, carnosus, expression
IdentifiersURN: urn:nbn:se:kth:diva-20974ISI: 000171237700038OAI: oai:DiVA.org:kth-20974DiVA: diva2:339671
QC 201005252010-08-102010-08-10Bibliographically approved