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Applied and Environmental Microbiology, October 2001, p. 4678-4684, Vol. 67, No. 10
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.10.4678-4684.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Generation of Metal-Binding Staphylococci through Surface Display of Combinatorially Engineered Cellulose-Binding Domains

Henrik Wernérus, Janne Lehtiö, Tuula Teeri, Per-Åke Nygren, and Stefan Ståhl^*

Department of Biotechnology, SCFAB, Kungliga Tekniska Högskolan, SE-10691 Stockholm, Sweden

Received 8 May 2001/Accepted 3 July 2001

Ni²⁺-binding staphylococci were generated through surface display of combinatorially engineered variants of a fungal cellulose-binding domain (CBD) from Trichoderma reesei cellulase Cel7A. 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.

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^* Corresponding author. Mailing address: Dept. of Biotechnology, SCFAB, Kungliga Tekniska Högskolan, SE-10691 Stockholm, Sweden. Phone: 46 8 55378329. Fax: 46 8 55378481. E-mail: stefans{at}biochem.kth.se.

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Applied and Environmental Microbiology, October 2001, p. 4678-4684, Vol. 67, No. 10
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.10.4678-4684.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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