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Applied and Environmental Microbiology, March 1999, p. 1092-1098, Vol. 65, No. 3
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Enhanced Bioaccumulation of Heavy Metal Ions by Bacterial Cells Due to Surface Display of Short Metal Binding Peptides

Pavel Kotrba,1 Lucie Dolecková,2 Víctor de Lorenzo,3 and Tomas Ruml1,*

Department of Biochemistry and Microbiology, Institute of Chemical Technology, 166 28 Prague,1 and Department of Biochemistry, Institute of Organic Chemistry and Biochemistry CAS, 166 10 Prague,2 Czech Republic, and Centro Nacional de Biotecnologia, CSIC, 28049 Madrid, Spain3

Received 7 August 1998/Accepted 18 December 1998

Metal binding peptides of sequences Gly-His-His-Pro-His-Gly (named HP) and Gly-Cys-Gly-Cys-Pro-Cys-Gly-Cys-Gly (named CP) were genetically engineered into LamB protein and expressed in Escherichia coli. The Cd2+-to-HP and Cd2+-to-CP stoichiometries of peptides were 1:1 and 3:1, respectively. Hybrid LamB proteins were found to be properly folded in the outer membrane of E. coli. Isolated cell envelopes of E. coli bearing newly added metal binding peptides showed an up to 1.8-fold increase in Cd2+ binding capacity. The bioaccumulation of Cd2+, Cu2+, and Zn2+ by E. coli was evaluated. Surface display of CP multiplied the ability of E. coli to bind Cd2+ from growth medium fourfold. Display of HP peptide did not contribute to an increase in the accumulation of Cu2+ and Zn2+. However, Cu2+ ceased contribution of HP for Cd2+ accumulation, probably due to the strong binding of Cu2+ to HP. Thus, considering the cooperation of cell structures with inserted peptides, the relative affinities of metal binding peptide and, for example, the cell wall to metal ion should be taken into account in the rational design of peptide sequences possessing specificity for a particular metal.


* Corresponding author. Mailing address: Department of Biochemistry and Microbiology, Institute of Chemical Technology, Technická 3, 166 28 Prague, Czech Republic. Phone: (420) 2-24353022. Fax: (420) 2-3119990. E-mail: TOMAS.RUML{at}VSCHT.CZ.


Applied and Environmental Microbiology, March 1999, p. 1092-1098, Vol. 65, No. 3
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



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